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abstract book

ABSTRACT BOOK
13TH INTERNATIONAL CONFERENCE OF THE
INTERNATIONAL MESOTHELIOMA INTEREST GROUP
Follow us on Twitter @iMig_meso
for updates during the Conference!
ABSTRACT BOOK
13TH INTERNATIONAL CONFERENCE OF THE I NTERNATIONAL MESOTHELIOMA INTEREST GROUP
Table of contents
PLENARY SESSIONS
Please note that the iMig 2016 Abstract Book only includes
Peer Reviewed Abstracts and excludes any abstracts from
Invited Speakers.
3
PL01: ORIGINS OF MESOTHELIOMA
MONDAY, MAY 2, 2016
08:25 – 10:25
PL02:
PREDICTING THE OUTCOME
MONDAY, MAY 2, 2016
11:15 – 12:45
PL03: THE KNIFE OR THE NEEDLE?
TUESDAY, MAY 3, 2016
09:00 – 10:30
[NO PEER REVIEW ABSTRACTS]
PL04: FROM GENETICS TO THERAPY
11:15 – 12:45
6
PL05:
WHAT IS IN THE LOCKER NOW?
WEDNESDAY, MAY 4, 2016
09:00 – 10:30
8
PL06:
THE “IMMUNE WAR” ON
MESOTHELIOMA
11:15 – 12:45
MINI SYMPOSIUM
3
4
11
12
MS01:
MARF INTERNATIONAL MESO UK
MINI SYMPOSIUM
MONDAY, MAY 2, 2016
14:15 – 15:45
MS02:
CELL DEATH MECHANISMS
MONDAY, MAY 2, 2016
14:15 – 15:45
MS03: IMAGING AND ENDPOINT EVALUATION
MONDAY, MAY 2, 2016
14:15 – 15:45
17
MS04: CELL AND VACCINE BASED THERAPY
MONDAY, MAY 2, 2016
14:15 – 15:45
22
MS05:
OPTIMUM DIAGNOSTIC PATHWAY FOR SUSPECTED MESOTHELIOMA
MONDAY, MAY 2, 2016
16:30 – 18:00
27
MS06:
ASBESTOS CONTROL
MONDAY, MAY 2, 2016
16:30 – 18:00
32
MS07:
BAP1 AND GENETICS
MONDAY, MAY 2, 2016
16:30 – 18:00
36
12
MS08:
PATHOLOGY
MONDAY, MAY 2, 2016
16:30 – 18:00
39
MS09:
SURGERY (TECHNICAL ASPECTS)
14:15 – 15:45
44
MS10:
NOVEL TARGETS ENTERING IN THE CLINIC
14:15 – 15:45
46
MS11:
CRITICAL SIGNALING PATHWAY
14:15 – 15:45
50
MS12: TREATMENT ADVANCES IN PERITONEAL
MESOTHELIOMA / PALLIATIVE CARE
FOR ALL MESOTHELIOMA
TUESDAY, MAY 3, 2016 14:15 – 15:45
55
MS13:
GENOMICS AND DRUG SENSITIVITY
16:30 – 18:00
57
MS14:
RADIOTHERAPY
16:30 – 18:00
60
MS15:
MULTIMODALITY
16:30 – 18:00
64
MS16:
NOVEL IMMUNE STRATEGIES
16:30 – 18:00
68
14
POSTER SESSIONS
73
PP01:
POSTER MIXER AND POSTER DISCUSSION SESSION 1
MONDAY, MAY 2, 2016
18:00 – 19:30
73
PP02:
POSTER MIXER AND POSTER DISCUSSION SESSION 2
18:00 – 19:30
117
AUTHOR INDEX
159
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ABSTRACT BOOK
PLENARY SESSIONS
PL01: ORIGINS OF MESOTHELIOMA
MONDAY, MAY 2, 2016
08:25 – 10:25
PL01.04: PLEURAL MESOTHELIOMA AND
ASBESTOS EXPOSURE: A CASE-CONTROL STUDY
WITH QUANTITATIVE RISK ASSESSMENT
Daniela Ferrante1, Dario Mirabelli2, Sara Tunesi3 , Benedetto
Terracini2, Corrado Magnani4
CPO-Piemonte and Unit of Medical Statistics and Epidemiology, Department of Translational Medicine, University of Eastern
Piedmont, Novara, ITALY, 2Center for Cancer Epidemiology and
Prevention, City of Health and Science Hospital; Human Genetics Foundation, HuGeF, Turin, ITALY,3CPO-Piemonte and Unit of
Medical Statistics and Epidemiology, Department of Translational
Medicine, University of Eastern Piedmont; Center for Cancer Epidemiology and Prevention, City of Health and Science Hospital,
Turin, ITALY, 4CPO-Piemonte and Unit of Medical Statistics and
Epidemiology, Department of Translational Medicine, University
of Eastern Piedmont; Human Genetics Foundation, HuGeF, Turin,
ITALY
1
Objectives: The area of Casale Monferrato (NW Italy, population around 100,000) showed an extremely high incidence of
malignant mesothelioma (MM) caused by the “Eternit” plant,
the most important asbestos cement plant in Italy active in the
town of Casale Monferrato for 80 years. During 1990-2010, the
annual incidence rate of definite pleural MM (excluding diagnoses of “probable” and “possible” MM) was 27.3 (per 100,000)
among men and 15 (per 100,000) among women, about 10
times higher than the corresponding Italian rate. Several studies have estimated the effect of asbestos exposure in this population considering the risk of MM separately for occupational,
environmental and domestic exposure. The purpose of the
present population-based case-control study was to quantify
the association between MM and asbestos cumulative exposure
using individual assessment of all sources of exposure.
Methods: The study included the incident cases of pleural malignant mesothelioma diagnosed from 1/1/2001 to 30/6/2006
to residents in the area. Cases were detected in the hospitals
of the area. The controls were a random sample of residents
matched to cases by sex and date of birth. Cases and controls
were interviewed with a standardized questionnaire including
sections on demographic characteristics, lifelong occupational
and residential histories, selected leisure time activities and
characteristics of the home environment possibly relevant for
asbestos exposure. Two hundred cases and 348 controls were
included in the study. Asbestos exposure was assessed by an
experienced rater and cumulative exposure was computed
considering all sources of exposure. The data analysis was
based on unconditional logistic regression adjusting all models
for gender, age at diagnosis and type of interview (subjects vs
with proxy).
Results: A highly statistically significant trend in the risk of
pleural malignant mesothelioma was observed with increasing
total (occupational and non occupational) cumulative exposure.
ORs increased from 4.4 (CI 95% 1.7 to 11.3) for cumulative
exposure <1 f/mL-years to 62.1 (CI 95% 22.2 to 173.2) for cumulative exposures above 10 f/mL-years when both occupational and non-occupational exposures were considered. Among 84
cases and 201 controls never occupationally exposed, corresponding ORs were 3.8 (CI 95% 1.3 to 11.1) and 23.3 (CI 95%
2.9 to 186.9) (reference: residents only exposed to background
levels of asbestos). Having a family member occupationally
exposed to asbestos doubles the risk of MM (38 cases and 35
controls; OR=2.2; CI95% 1.2-4.0). Having a garden or courtyard
paved with asbestos cement tailings, an asbestos cement roof
or buildings near home were also associated with a significant increase in the OR (152 cases and 221 controls; OR=1.9
CI95%1.2-3.0).
Conclusion: This study underlines that, in addition to occupational exposures, environmental and familial/domestic exposures to asbestos also contribute to the occurrence of MM in
the population of Casale Monferrato. Continuing epidemiological surveillance and investigation into the specific routes and
circumstances of exposures contributing to MM occurrence in
this population is, therefore, important.
Keywords: pleura, asbestos, environmental asbestos exposure, cumulative exposure
PL01.06: LONG-FIBRE CARBON NANOTUBES
INDUCE PLEURAL MESOTHELIOMA VIA
SILENCING AND/OR LOSS OF KEY TUMOUR
SUPPRESSOR GENES
Tatyana Chernova1, Fiona A. Murphy1, Sara Galavotti1, XiaoMing Sun1, Ian R. Powley1, Stefano Grosso1, Anja Schinwald2,
David Dinsdale1, John Le Quesne1, Jonathan Bennett3 , Apostolos Nakas3 , Peter Greaves4 , Craig A. Poland5, Ken Donaldson2,
Martin Bushell1, Anne E. Willis1, Marion Macfarlane1
MRC Toxicology Unit, Leicester, UNITED KINGDOM, 2Centre For
Inflammation Research, MRC/University of Edinburgh, Edinburgh,
UNITED KINGDOM, 3Glenfield Hospital, UHL NHS Trust, Leicester,
UNITED KINGDOM, 4Department of Cancer Studies, University of
Leicester, Leicester, UNITED KINGDOM, 5Institute of Occupational
Medicine, Edinburgh, UNITED KINGDOM
1
Objectives: Exposure to asbestos fibres causes pathological
changes in the pleural cavity including malignant mesothelioma. Length-dependent retention of asbestos fibres in the
pleural cavity is crucial for disease development. Chronic
inflammation induced by pathogenic asbestos fibres plays a
key role in carcinogenesis and epigenetic events, rather than
driver mutations, are considered to be major causative factors.
Manufactured carbon nanotubes (CNT) are similar to asbestos
in terms of their high aspect ratio and thus may pose an asbestos-like inhalation hazard, however the molecular mechanisms
underlying their carcinogenic potential have not been sufficiently explored.
Methods: Using a model of direct injection into the pleural
cavity we compared the molecular changes which occur at the
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ABSTRACT BOOK
mesothelium after exposure to short and long asbestos fibres
and short and long CNT over 1 year following injection.
Results: We show a common pro-oncogenic activity of long
CNT and long asbestos throughout disease progression. The
common key molecular events encompass changes in gene
expression and signaling pathway activation, oxidative DNA
damage, increased mitosis and proliferation. Instillation of long
CNT into the pleural cavity of mice induces chronic inflammation and pro-oncogenic changes leading to development
of mesothelioma with deletion of p19/Arf and silencing of p16/
Ink4a and NF2. Epigenetic changes induced by pathogenic fibres
occur at the pre-neoplastic stage of disease and may play a key
role in progression of pleural inflammatory lesions to malignant
mesothelioma.
Conclusion: Together these data demonstrate that exposure
to long CNT induces development of pleural mesothelioma
replicating the pathogenesis of human disease and highlights
commonality in the hazard mechanism of long pathogenic
fibres at the molecular level. Crucially, our findings reinforce
concerns that long CNT may pose an asbestos-like hazard,
leading to malignant mesothelioma.
enhances the expression of genes encoding cell-cycle promoting proteins including CCDN1 and FOXM1 and connective
tissue growth factor (CTGF), the latter of which was associated
with extracellular matrix formation of the MM cells in vivo.
To determine the biological roles of YAP on untransformed
mesothelial cells, we established immortalized mesothelial cell
lines (HOMC), and examined whether YAP activation induces
malignant phenotypes in the cells. We found that transduction
of both wild-type and constitutively active-type (S127A) YAP enhanced HOMC-cell proliferation in vitro. We also demonstrated
that YAP-transduced HOMC cells showed enhanced tumorigenicity in vivo after inoculation into nude mice subcutaneously
or into intrathoracic cavities. Finally, we are currently analyzing
whether or not TAZ, a paralog of YAP, is also involved in the
dysregulation of the YAP/TAZ target gene expressions in MM
cells, and YAP/TAZ activation is a synergistic effect on MM cell
proliferation and promotion.
Conclusion: Our results indicate that the NF2-Hippo pathway
inactivation induces YAP and TAZ activation, which confers
more malignant phenotypes of mesothelial cells.
Keywords: signal transduction, Hippo pathway
Keywords: in vivo model, epigenetics, carbon nanotubes,
mesothelioma
PL02: PREDICTING THE OUTCOME
MONDAY, MAY 2, 2016
11:15 – 12:45
PL01.09: CONSTITUTIVE YAP ACTIVATION
INDUCES MALIGNANT PHENOTYPES OF
IMMORTALIZED MESOTHELIAL CELLS
Yoshitaka Sekido
Division of Molecular Oncology, Aichi Cancer Center Research
Institute, Nagoya, JAPAN
Objectives: Malignant mesothelioma (MM) is an aggressive tumor arising primarily from pleural or peritoneal cavities, which
is caused by asbestos exposure after long latency. Three major
tumor suppressor genes which are frequently mutated in MM
are CDKN2A, NF2, and BAP1. NF2 encodes Merlin, a member of
the Ezrin-Radixin-Moesin protein, and Merlin regulates the Hippo signaling pathway, which has been shown to play important
roles in organ size control and cancer development. Inactivation
of Hippo pathway is known to induce constitutive underphosphorylation/activation of YAP transcriptional coactivator. The
objective of this study is to demonstrate whether YAP confers
more malignant phenotypes to mesothelial cells in vitro and in
vivo.
Methods: Immortalized mesothelial cell lines (HOMC) were established by transduction of HPV-E6/E7 and hTERT into primary
mesothelial cells. Immortalized mesothelial cells which were
transduced with YAP or control vectors were injected subcutaneously or into the right thoracic cavity of nude mice.
Results: Our previous studies have identified alteration or aberrant expression of the components in this cascade including
LATS2, SAV1 and AJUBA, which causes constitutive activation
of YAP transcriptional coactivator. We found that activated YAP
PL02.03: IMPACT OF TUMOR THICKNESS ON
SURVIVAL AFTER ACCELERATED HEMITHORACIC
RADIATION FOLLOWED BY EXTRAPLEURAL
PNEUMONECTOMY
Marc De Perrot, Ronald Feld, Penelope Bradbury, Natasha
Leighl, Bc John Cho
Thoracic Surgery, Toronto General Hospital and Princess Margaret Cancer Center, Toronto, ON, CANADA
Objectives: Surgery for mesothelioma after radiation therapy
(SMART) provides encouraging results in patients with malignant pleural mesothelioma of epithelial subtype. However,
patient selection for this approach based on radiological parameters remains not well defined. In this analysis, we reviewed
the impact of tumor thickness (TT) on long-term outcome in an
attempt to find radiological criteria to select patients for this
therapy.
Methods: Pre-treatment CT scan was reviewed for all patients undergoing the SMART approach between 10/2008 and
11/2015. Patients undergoing chemotherapy before SMART
were excluded from analysis (n=4). The thickest part of the tumor was measured on three sites along the chest wall (anterior,
middle, posterior), mediastinum (upper, lower anterior, lower
posterior) and diaphragm (anterior, middle, posterior) using
modified RECIST criteria. TT of <1cm, 1-1.5cm and >1.5cm was
then used as a cut-off. All patients were followed up until death
or 11/2015. Survival was calculated from the start of radiation
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ABSTRACT BOOK
using life table analysis and statistical differences were determined by log-rank test.
Results: All 70 consecutive patients (81% men, age 41-82,
median 64 years) included in the SMART protocol completed
radiation (25-30 Gy in 5 fractions) and surgery (extrapleural
pneumonectomy) with no 30-day mortality. Two patients died
after discharged from hospital for a treatment related mortality of 2.9%. The overall median survival and disease-free
survival (DFS) reached 45 months and 31 months in patients
with epithelial subtypes, respectively, compared to 13 months
(p=0.0008) and 8 months (p=0.0002) in patients with biphasic
subtypes. Among patients with epithelial subtype, the 5-year
survival reached 51±13% in the absence of pT4N1-2 disease
on final pathology (median survival not reached after a median
follow-up of 29 months), while the median survival was 19
months in pT4N1-2 disease (p=0.006). The median DFS was 16
months in patients with epithelial pT4N1-2 disease compared
to 48 months in the remaining patients with epithelial subtype (p=0.01). Age (p=0.3) and gender (p=0.2) did not impact
survival and DFS. Maximal TT of <1cm, 1-1.5cm and >1.5cm
on the chest wall had no impact on survival (p=0.6) and DFS
(p=0.8). Maximal TT <1cm on the mediastinum was associated
with better DFS (p=0.03), but had no significant impact on
survival (p=0.1). In contrast, maximal TT on the diaphragm had
a major impact on survival and DFS. Median survival reached
51 months when maximal TT was <1cm on the diaphragm, 24
months when maximal TT was 1-1.5cm and 12 months when
maximal TT was >1.5cm (p<0.0001). All patients had recurrence
within 18 months when maximal TT on the diaphragm was
>1.5cm (median DFS 10 months), while the 3-year DFS reached
54±12% (median DFS, 47 months) when maximal TT was <1cm
and 36±12% (median DFS, 14 months) when maximal TT was
1-1.5cm (p<0.0001).
Conclusion: The outcome of patients with epithelial subtypes
remains encouraging after the SMART approach, particularly in
the absence of pT4N1-2 disease. Maximal TT on the diaphragm
based on pre-treatment CT scan appears to be a good predictor
of outcome, independently of histologic subtypes, and could
potentially be used as a selection criteria for this approach.
PL02.05: MUTATION PROFILES OF MALIGNANT
PLEURAL MESOTHELIOMAS ACCORDING TO
MOLECULAR CLASSIFICATION
Lisa Quetel1, Clément Meiller1, Robin Tranchant1, Annie Renier1, Françoise Galateau-Sallé2, Marie-Christine Copin3 , Paul
Hofman4 , Françoise Le Pimpec-Barthes1, Sandrine Imbeaud1,
Jessica Zucman-Rossi1, Marie Claude Jaurand1, Didier Jean1
Inserm U.1162, INSERM U.1162, Paris, FRANCE, 2MESOBANK, Lyon,
FRANCE, 3CHRU Lille, Lille, FRANCE, 4CHU Nice, Nice, FRANCE
1
Objectives: Development of precision medicine for Malignant
Pleural Mesothelioma (MPM) needs a deep knowledge of the
molecular changes associated with mesothelial carcinogenesis
especially to take into account the tumor variability between patients. Recently, based on transcriptomic data, we defined a ro-
bust molecular classification of MPM composed of two groups,
C1 and C2, linked to histology and survival. C1 MPM exhibited
more frequent BAP1 alterations. To better define the mutation
profile of the C1 and C2 MPM groups, we performed targeted
Next-Generation Sequencing (NGS) of candidate genes.
Methods: NGS (Miseq, Illumina) was performed using 165
MPM including 60 MPM cultures and 105 frozen MPM tumors
samples. Twenty-two candidate genes were selected: key
altered genes in mesothelial carcinogenesis (CDKN2A, CDKN2B, NF2, BAP1, TP53 and LATS2) previously sequenced by
Sanger method, genes mutated at low frequency in MPM
(KRAS, HRAS, EGFR, CTNNB1…) and genes recently reported
as altered in MPM (CUL1, ARID1A, ARID2, SMARCA4, SETD2…).
The TERT promoter, in which we previously identified oncogenic hot-spot mutations, was also included. Genetic alterations
were analyzed with a total depth of 200X. Large deletions were
detected from NGS data based on coverage and confirmed by
PCR on genomic DNA or by Multiplex ligation-dependent probe
amplification (MLPA). Gene expression was also assessed by
RT-qPCR to validate absence of transcript for genes with large
deletion.
Results: Data demonstrated an enrichment in C>T transitions
and high frequency of large biallelic deletion in CDKN2A/
CDKN2B, NF2 and BAP1 genes. Previous genetic alterations
identified by Sanger sequencing in our collection have been
found. Variants inducing protein structure modification and
not identified as SNP, were found in 19 genes. The mutation
frequencies are consistent with literature data for previously
well-characterized genes and allow to precise the frequency
for the others. Genetic alterations deleterious to the function
of the protein (deletion/insertion, splice-site mutation, substitutions nonsense and damaging missense predicted by SIFT
and Polyphen) represent more than 60% of the non-synonymous variants, and are enriched in 10 genes. Among them,
the highest alteration frequency was found in CDKN2A, CDKN2B, NF2 and BAP1 genes (over 25%) and frequencies around
5-10% were found in TP53, LATS2, and SETD2 genes. TERT promoter mutations were found at an overall rate of 18% in all
MPM and 59% in sarcomatoid MPM, supporting the strong
association with this histological subtype (P=0.0001), which
we described in a precedent study. The highest frequency
of BAP1 mutations in C1 MPM (P=0.0006) was confirmed.
Moreover, an enrichment in SETD2 (histone methyltransferase)
mutations and a significant association with ARID1A (member of
the SWI/SNF chromatin remodeling family) mutations, mainly
consisting in non-deleterious substitutions, were identified.
Conversely, relevant mutations in TP53 gene were significantly
associated with C2 MPM (P =0.02).
Conclusion: The NGS gene candidate approach precise the
genetic landscape of C1 and C2 MPM main tumor groups. C1
MPM are characterized by mutations in genes involved in chromatin organization. Interestingly, mutations in TP53, which is
linked with tumor aggressiveness, are found mainly in the MPM
of the C2 group, gathering patients with the worse prognosis.
Keywords: Tumor molecular classification, Genetic alterations,
Next Generation Sequencing (NGS)
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ABSTRACT BOOK
PL04.03: PHASE 1 STUDY OF TAZEMETOSTAT
(EPZ-6438), AN INHIBITOR OF EZH2, IN PATIENTS
WITH NON-HODGKIN LYMPHOMA AND SOLID
TUMORS
occurring in >10% of pts were: asthenia, anorexia, thrombocytopenia, nausea, constipation, diarrhea, vomiting and muscle
spasms. Four pts had grade ≥3 treatment related AE’s: thrombocytopenia, neutropenia, hypertension, and transaminase
elevation. The recommended phase 2 dose was determined to
be 800 mg BID. Of 16 response-evaluable NHL pts, objective
responses (CRs, PRs) were observed in 56% (5/10 DLBCL, 3/5
FL and 1/1 MZL). Four NHL pts remained on study for >1 year.
Of the ST pts, all 6 who experienced tumor reduction had either
INI1- or SMARCA4-negative tumors as of 31-Aug. 2015. This
includes a CR that is ongoing through 65 weeks (MRT), PRs
(MRT, ES, MRT of ovary) and stable disease >24 weeks (ES,
MRT of ovary).
Vincent Ribrag1, Antoine Italiano2, Jean-Charles Soria1,
Jean-Marie Michot1, Anna Schmidt2, Sophie Postel-Vinay1,
Fontanet Bijou3 , Jean-Michele Coindre2, Maud Toulmonde2,
Christophe Massard1, Stephen J. Blakemore4 , Alice Mcdonald4 ,
Scott Ribich4 , Blythe Thomson4 , Heike Keilhack4 , Maria Roche4 ,
John Larus4, Peter T. Ho4
Conclusion: Tazemetostat demonstrates a safety profile
favorable for chronic dosing and objective responses in pts with
relapsed or refractory B-cell NHL including DLBCL, FL and MZL
and in subjects with advanced STs consisting of MRT, ES, and
MRT of ovary. Phase 2 trials in B-cell NHL and INI1- or SMARCA4-negative tumors are enrolling. A phase 2 trial in pts with
BAP1-mutated mesothelioma is planned.
PL04: FROM GENETICS TO THERAPY
11:15 – 12:45
Institut Gustave Roussy, Villejuif, FRANCE, 2Institut Bergonie, Bordeaux, FRANCE, 3French Blood Institute, Bordeaux,
FRANCE, 4Epizyme Inc., Cambridge, MA, UNITED STATES OF
AMERICA
1
Objectives: The histone methyl transferase EZH2 is the catalytic subunit of the polycomb repressive complex 2 (PRC2) and
responsible for methylation of lysine 27 of histone H3 (H3K27),
which results in chromatin remodeling and repressed transcription when trimethylated. Aberrant EZH2 activity has been implicated as an oncogenic driver in non-Hodgkin lymphoma (NHL).
The SWI/SNF complex also remodels chromatin, activates
transcription and acts in opposition to PRC2. Oncogenesis from
mutation and/or loss of the SWI/SNF subunit INI1 in cancers
such as malignant rhabdoid tumor (MRT) is sensitive to EZH2
inhibition. Tazemetostat is a potent, selective small molecule inhibitor of EZH2 in phase 2 clinical development. EZH2 inhibition
may have therapeutic potential in BAP1 mutated mesothelioma
(Levine, Nat Med 2015).
Methods: This phase 1 open-label first-in-human study
evaluated the safety, tolerability, and preliminary efficacy of
tazemetostat administered orally as a monotherapy twice a day
(BID). Eligible patients (pts) had either a relapsed/refractory
B-cell NHL or solid tumors (ST). Archival tumor tissue from
NHL pts was analyzed for EZH2 hot spot mutations by either
amplicon-based next generation sequencing [NGS] or cobas®
EZH2 Mutation Test [in development]. In addition, cell-of-origin
in Diffuse Large B-cell Lymphoma (DLBCL) pts was determined
by immunohistochemistry using the Hans algorithm. For tumors
that were INI1-negative, central confirmation of diagnostic pathology and INI1 loss was performed. Tazemetostat was administered to subjects in 5 dose cohorts (100 mg, 200 mg, 400 mg,
800 mg and 1600 mg) and in 2 clinical pharmacology sub-study
cohorts. Tumor response assessments were performed every 8
weeks and graded according to Cheson/IWG criteria or RECIST
as appropriate.
Results: As of 7-Nov. 2015, 58 pts were enrolled to this trial
including 21 NHL pts, (14 DLBCL, 6 follicular lymphoma [FL]
and 1 marginal zone lymphoma [MZL]). Of the 37 ST pts, 8 had
INI1-negative tumors (MRT [5], epithelioid sarcoma (ES) [3])
and 3 had SMARCA4-negative tumors (MRT of ovary [2], thoracic sarcoma [1]). Adverse events (AE) regardless of attribution
Keywords: EZH2, Phase 1, Lymphoma, INI1
PL04.05: DIFFERENTIAL RESPONSE OF
MALIGNANT PLEURAL MESOTHELIOMA CELLS
TO YAP TARGETED THERAPY ACCORDING TO
MOLECULAR CLASSIFICATION
Robin Tranchant1, Annie Renier1, Lisa Quetel1, Leanne De
Koning2, Françoise Le Pimpec-Barthes1, Jessica Zucman-Rossi1,
Marie Claude Jaurand1, Didier Jean1
Inserm U.1162, INSERM U.1162, Paris, FRANCE, 2Rppa Platform,
Institut Curie, Paris, FRANCE
1
Objectives: Novel target therapies require better knowledge
of molecular and clinico-biological heterogeneity of tumors. To
better characterize MPM heterogeneity, we recently identified
a robust MPM transcriptomic classification defining two groups
(C1 and C2). Epithelioid MPM, the most frequent histologic
subtype, was found in both tumor groups, with a worse survival
prognosis in the C2 group. These groups differ by their mutation profile and by specifically deregulated pathways such as
epithelial-mesenchymal transition (EMT) and TGFβ pathway.
The aim of the work was to determine if the effect of a panel of
ten specific molecular inhibitors induced a differential response
in the 2 groups. Both inhibitors of epigenetic regulation and
signaling pathways involved in mesothelial carcinogenesis were
investigated.
Methods: A panel of 18 MPM primary cultured cells classified
in C1 or C2 and characterized for genetic alteration in genes
involved in mesothelial carcinogenesis. Cell viability was
determined by the MTS assay after treatment with a gradient
concentration of ten inhibitors for 48 hours (Table 1). Gene
expression was measured by quantitative RT-PCR, and protein
phosphorylation and expression by Reverse Phase Protein
Array.
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ABSTRACT BOOK
Results: The mTOR/PI3K/Akt pathway inhibitor (PF_04691502),
YAP/TEAD association inhibitor (Verteporfin) and the histone
deacetylase inhibitor (Vorinostat) were the most effective to
reduce MPM viability (Table 1). MPM of the C1 molecular group
were more sensitive to Verteporfin treatment than MPM of the
C2 group (P=0.01 – Figure 1). YAP activity is known to be regulated by Hippo pathway, but
no significant relationship between Verteporfin sensitivity
and the mutations status of two members of Hippo pathway, NF2 and LATS2, was observed. A decrease in YAP phosphorylation (P=0.04) and an overexpression of YAP target
genes (CTGF and CYR61, P<0.01) were observed in C2 MPM in
comparison with C1 MPM, indicating a higher co-transcriptional
activity of YAP in C2 MPM. CTGF mRNA expression was predictive of Verteporfin sensitivity. Verteporfin induced a downregulation of YAP target genes (CTGF and CYR61), but also of target
genes of TGFβ pathway (MMP2 andSERPINE1).
Conclusion: MPM sensibility to Verteporfin was dependent of
YAP activity and could be predicted by CTGF gene expression.
Our data suggest that YAP deregulation is stronger in C2 group
and is not only associated with Hippo inactivation. YAP targeting by Verteporfin may be a promising strategy for MPM patient
treatment, especially for MPM of the C1 molecular group.
Keywords: Tumor molecular classification, Genomics, Drug
sensitivity, Hippo pathway
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PL04.06: NOVEL SYNERGISTIC CELL THERAPIES
FOR THE TREATMENT OF MALIGNANT PLEURAL
MESOTHELIOMA
Keywords: chemotherapy, MSCTRAIL, mesothelioma, Cell
Therapy
Beth Sage, Krishna Kolluri, Zhenqiang Yuan, Neelam Kumar,
Adam Giangreco, Sam Janes
PL05: WHAT IS IN THE LOCKER NOW?
09:00 – 10:30
Ucl Respiratory, University College London, London, UNITED
KINGDOM
Objectives: Malignant pleural mesothelioma (MPM) is an
aggressive fatal cancer with no effective treatments. Mesenchymal stem cells (MSCs) migrate and incorporate into
tumour stroma making them good vehicles for the delivery of
anti-cancer therapies. TNF-related apoptosis inducing ligand
(TRAIL) selectively induces apoptosis in malignant cells without
affecting healthy tissues and is known to target the extrinsic
apoptotic pathway. However, some cancer cells are resistant
to TRAIL due to expression of proteins that block this pathway.
Current chemotherapeutics target the proteins that inhibit the
extrinsic apoptotic pathway suggesting that MSCTRAIL could be
used in conjunction with those agents to increase the therapeutic effect. This study aimed to test whether MSCs modified to
express TRAIL (MSCTRAIL) alone or in conjunction with Vorinostat (cFLIP inhibitor), LCL161 (IAP inhibitor), SNS032 (cFLIP
& MCL1 inhibitor) and Obatoclax BCL2 family inhibitor) could
be a successful treatment for MPM.
Methods: Human MSCs were transduced with a lentiviral
vector containing TRAIL. The biological activity of MSCTRAIL
was determined using co-culture experiments where DiI stained
MPM cells were incubated in a 4:1 ratio with MSCTRAIL cells
with or without chemotherapy or rTRAIL for 24 hours. Apoptosis and cell death were determined using Annexin V and DAPI
staining on flow cytometry. To test the effect of MSCTRAIL in
vivo a bioluminescent tumour model was established. MPM
cells were transduced with a luciferase lentivirus and injected
into the pleural cavity of NOD/SCID mice to establish an orthotopic tumour model. MSCTRAIL cells were delivered via tail vein
injections on days 5, 9, 12, 15 and 18 post tumour inoculation
and bioluminescence was measured twice weekly.
Results: MSCs were successfully transduced with TRAIL with
96% efficiency and TRAIL production was confirmed by ELISA.
Ten human MPM cell lines were tested with 6 being sensitive to
TRAIL and 4 resistant. In vivo delivery of MSCTRAIL to xenograft
tumours from a TRAIL sensitive cell line resulted in a significant
reduction in MPM tumour growth. TRAIL resistant cell lines
were further tested with a combination of MSCTRAIL and 4 different chemotherapy agents and showed a significant increase
in cell death. 3 cell lines were sensitive to all chemotherapeutic
agents in combination with MSCTRAIL whilst one was sensitive
to SAHA, SNS032 and LCL161 with MSCTRAIL but not Obatoclax.
Conclusion: MSCs can be successfully transduced with TRAIL
and induce apoptosis and death of MPM cells in vitro. Intravenous delivery of MSCTRAIL causes a significant reduction in
TRAIL sensitive MPM tumour growth and resistant cells can be
made sensitive by the addition of agents that target different
elements of the extrinsic apoptotic pathway. MSCTRAIL is a
potential novel cellular therapy for this currently untreatable
disease.
PL05.01: PHASE II STUDY ON INTENSITY
MODULATED PLEURAL RADIATION THERAPY
(IMPRINT) FOR MALIGNANT PLEURAL
MESOTHELIOMA: FINAL RESULTS
Andreas Rimner1, Marjorie G. Zauderer2, Daniel R. Gomez3 ,
Prasad S. Adusumilli4 , Preeti Parhar1, Kaitlin M. Woo5, Ronglai
Shen5, Michelle Ginsberg6 , David Rice7, Anne Tsao8 , Kenneth
E. Rosenzweig9, Abraham J. Wu1, Ellen Yorke10, Valerie Rusch11,
Lee Krug12
Radiation Oncology, Memorial Sloan Kettering Cancer Center,
New York City, NY, UNITED STATES OF AMERICA, 2Medicine, Memorial Sloan Kettering Cancer Center, New York, UNITED STATES
OF AMERICA, 3Radiation Oncology, MDACC, Houston, UNITED
STATES OF AMERICA,4Surgery, Memorial Sloan Kettering Cancer
Center, New York City, NY, UNITED STATES OF AMERICA, 5Biostatistics, Memorial Sloan Kettering Cancer Center, New York City,
NY, UNITED STATES OF AMERICA, 6Radiology, Memorial Sloan
Kettering Cancer Center, New York City, NY, UNITED STATES OF
AMERICA, 7Surgery, MDACC, Houston, TX, UNITED STATES OF
AMERICA, 8Md Anderson Cancer Center, The University of Texas,
Houston, TX, UNITED STATES OF AMERICA, 9Radiation Oncology,
Mount Sinai Medical Center, New York, NY, UNITED STATES OF
AMERICA, 10Medical Physics, Memorial Sloan Kettering Cancer
Center, New York City, NY, UNITED STATES OF AMERICA, 11Surgery, Memorial Sloan Kettering Cancer Center, New York, NY,
UNITED STATES OF AMERICA, 12Bristol Myers Squibb, New York,
UNITED STATES OF AMERICA
1
Objectives: Adjuvant radiation therapy for malignant pleural
mesothelioma (MPM) is particularly challenging in patients
with two intact lungs after pleurectomy/decortication (P/D) or
those with unresectable disease due to the risk for radiation
pneumonitis (RP). Here we report the final results of a prospective phase II study to determine the safety of hemithoracic
pleural IMRT as part of a multimodality lung-sparing treatment
approach.
Methods: Patients received up to 4 cycles of pemetrexed/platinum chemotherapy. If feasible, P/D was performed. Hemithoracic pleural IMRT was administered in 28 fractions for a total
planned dose consistent with normal tissue constraints, up to
5040 cGy, as previously described (Rosenzweig et al., IJROBP
2012). The primary endpoint was the incidence of ≥grade 3 RP
defined per Common Terminology Criteria for Adverse Events,
v4.0. Steroids, typically 40mg prednisone, were rapidly initiated
for ≥grade 2 RP. A Simon two-stage design was used with a
safety analysis after the first 9 patients. As only one patient developed ≥grade 3 RP over 4 months, the cohort was expanded
to 27 evaluable patients, defined as having initiated RT.
iMig2016.ORG
8
ABSTRACT BOOK
Results: Forty-five patients were enrolled. The median age was
68 years (range 38-79). Median KPS was 90% (range 70-100%).
Ten patients had sarcomatoid or biphasic and 35 had epithelioid MPM. Sixty percent had advanced stage III/IV MPM with
49% deemed unresectable. 18 patients came off study prior to
receiving IMRT (9 due to disease progression, 5 due to patient
refusal of surgery or RT, 2 for a change in the planned surgical procedure to extrapleural pneumonectomy, and 2 due to
complications from chemotherapy). 27 patients initiated IMRT
[median dose 4680cGy (range 2880 to 5040cGy)]; one patient
had distant disease progression after 16 fractions; all other
patients completed their radiation treatment as planned. Eight
patients (30%) developed ≥grade 2 RP: 6 patients experienced
grade 2 RP with symptom improvement after steroid initiation.
Only 2 patients experienced grade 3 RP and were successfully
weaned from oxygen after a course of steroids. Other ≥grade
2 radiation-related toxicities included fatigue (41%), nausea
(41%), esophagitis (30%), and cough (11%). No grade 4 or 5
radiation-related toxicities were observed. The median progression-free and overall survival (OS) was 12.4 and 23.7 months,
with a 2-year OS of 59% in resectable and 25% in unresectable
patients.
Conclusion: Hemithoracic pleural IMRT has an acceptable toxicity profile. Early intervention with steroids appears effective in
avoiding severe toxicities of RP. Survival rates of our lung-sparing multimodality regimen were promising for this advanced
patient population. This novel radiation technique will be further
tested in a multicenter safety study to establish its exportability
for the treatment of locally advanced MPM.
Keywords: pleural IMRT, pleurectomy/decortication, trimodality therapy, radiation pneumonitis
the ipsilateral, contralateral and total lung volumes, all minus
GTV, were exported for analysis. The maximum RP grade (Common Terminology Criteria for Adverse Events, V4.0), onset time
after treatment start, disease laterality, age, gender and smoking history were obtained from patient records. Correlation of
categorical variables with Grade 2 or higher (G2+) and G3+ RP
was analyzed with Fishers’ exact test. Preliminary analysis of
RP correlation with mean organ dose and percent organ volume
receiving dose D (VD) at selected doses was analyzed with the
rank-sum test and the Cox model was used for detailed analysis
of VD with D in 2 Gy increments.
Results: 27 patients had G2+ RP: 13 had Grade 2, 11 Grade 3,
2 Grade 4, 1 Grade 5. Median prescription dose was 46.8 Gy
(range 39.6-50.4 Gy), all in 1.8 Gy fractions. Median age was
67.6 y (42-83). There were 79 males, 24 females; 63 patients
were former or current smokers, 40 were never-smokers; 44 patients had left-sided disease, 59 had right-sided. No categorical
variables were significantly correlated with RP, but there was a
trend (p=0.09) for RP3+ more likely for left-sided disease. The
Cox model analysis revealed significant correlation (p<0.05)
of RP2+ with total lung VD from 12 to 16 Gy, ipsilateral lung
VD from 38-44 Gy and heart VD from 36-48 Gy. The best p-values for heart VD were an order of magnitude smaller than those
for lung. The rank-sum analysis showed significant correlation
of G2+ and G3+ RP with mean heart dose and of G3+ with heart
V40.
Conclusion: In addition to radiation dose to the lungs, radiation dose to the heart is correlated with symptomatic RP in
this large cohort of MPM patients with two lungs treated with
hemithoracic pleural IMRT. Heart dose should be kept as low as
possible while maintaining target coverage. Continuing analysis
will determine specific planning constraints.
Keywords: IMRT, radiation pneumonitis, radiotherapy
PL05.02: PNEUMONITIS PREDICTORS IN
INTENSITY MODULATED RADIATION TREATMENT
OF MESOTHELIOMA PATIENTS WITH TWO LUNGS
Ellen Yorke1, Anthonia Ojo2, Andrew Jackson3 , Licheng Kuo1,
Ming Yan1, Andreas Rimner2
Medical Physics, Memorial Sloan Kettering Cancer Center, New
York City, NY, UNITED STATES OF AMERICA, 2Radiation Oncology, Memorial Sloan Kettering Cancer Center, New York City,
NY, UNITED STATES OF AMERICA, 3Medical Physics, Memorial
Sloan Kettering Cancer Center, New York, NY, UNITED STATES OF
AMERICA
1
Objectives: To determine dose-volume and clinical metrics
that correlate with symptomatic radiation pneumonitis (RP) in
malignant pleural mesothelioma (MPM) patients with two intact
lungs uniformly treated with hemithoracic pleural intensity
modulated radiation therapy (IMRT).
Methods: The treatment plans of 103 consecutive MPM
patients treated between 2/2005 and 3/2015 and satisfying the
above criteria were recalculated with a superposition-convolution algorithm. The treatments were designed to give the highest prescription < 50.4 Gy that satisfied departmental normal
tissue constraints. Dose-volume histograms for the heart and
PL05.05: RANDOMIZED PHASE II STUDY OF
ADJUVANT WT1 VACCINE FOR MALIGNANT
PLEURAL MESOTHELIOMA (MPM) AFTER
MULTIMODALITY THERAPY
Marjorie G. Zauderer1, Tao Dao1, Valerie Rusch2, Michelle Ginsberg3 , Anne Tsao4 , Katherine Panageas3 , David Scheinberg3 ,
Lee Krug1
Medicine, Memorial Sloan Kettering Cancer Center, New York,
UNITED STATES OF AMERICA, 2Surgery, Memorial Sloan Kettering Cancer Center, New York, UNITED STATES OF AMERICA, 3Memorial Sloan Kettering Cancer Center, New York, UNITED STATES
OF AMERICA, 4Md Anderson Cancer Center, The University of
Texas, Houston, TX, UNITED STATES OF AMERICA
1
Objectives: The WT1 gene product is highly expressed on
tumor cells of numerous cancers and nominally expressed on
normal adult tissues. This makes WT1 an ideal candidate for
a tumor selective cancer vaccine in malignancies that express
WT1, such as mesothelioma. Using four native and synthetic
peptide sequences from WT1, a multivalent peptide vaccine
was created to stimulate both CD4 and CD8 T cell responses.
iMig2016.ORG
9
ABSTRACT BOOK
Affinity was optimized and the peptides were combined with
Montanide adjuvant and co-administered with GM-CSF injected
locally. In a pilot trial including patients with previously treated
MPM, the vaccine was well-tolerated and CD4/8 immune
responses were generated. Subsequently, we began this
randomized, double-blind, placebo-controlled, phase II study
of the WT1 vaccine in patients with MPM expressing WT1 who
completed multimodality therapy.
Methods: After surgical resection as well as chemotherapy
and/or radiation, patients were randomly assigned to receive
Montanide and GM-CSF with or without the WT1 peptide
vaccine. Treatment consisted of 6 subcutaneous vaccinations
(Montanide, or Montanide with WT1 peptide vaccine) on weeks
0, 2, 4, 6, 8, and 10 beginning within 12 weeks after completing
multimodality therapy. Injection sites were prestimulated with
GM-CSF on days -2 and 0. Immune responses were evaluated
by ELISPOT and T-cell proliferation assays at week 12. Patients
were followed for progression with imaging every 3 months with
a primary endpoint of 1-year progression-free survival (PFS).
Results: 40 patients were randomized (21 on the placebo arm
and 19 with the complete WT1 vaccine) from two institutions.
Patient characteristics were well balanced between the arms,
with a majority of men and epithelioid histology. There were no
serious treatment related adverse events. Based on a pre-specified futility analysis of each arm (futility = 10 or more of the first
20 patients experience progression within 1 year), the placebo
control arm was closed in May 2015; accrual was stopped and
the vaccine arm was closed in November 2015. Median PFS
from randomization was 11.4 months (95% CI 4.4-24.3) in the
WT1 vaccine arm versus 5.7 months (95% CI 2.7-14.6) in the
placebo control arm (hazard ratio 0.69, p=0.3). Similarly, median overall survival (OS) from randomization was 21.4 months
(95% CI 8.5-40.4) in the WT1 vaccine arm versus 16.6 months
(95% CI 7.7-24.8) in the placebo control arm (hazard ratio 0.52,
p=0.14). In the subgroup with R0 resection, median OS was
39.3 months in the vaccine arm and 24.8 in the control arm
(p=0.04). PFS and OS were also examined in various subgroups
related to their immune response.
Conclusion: This randomized, controlled phase II trial demonstrated that administration of this analog WT1 peptide vaccine
in MPM was associated with a trend toward improved PFS and
OS, though the trial was not originally powered to determine
this effect. These results warrant additional better powered,
randomized studies to define the optimal use and benefit of this
vaccine in the treatment of mesothelioma. Supported by the
Department of Defense, the Mesothelioma Applied Research
Foundation, the National Cancer Institute, and Sellas Life
Sciences Group.
Keywords: Immunotherapy, WT1, adjuvant, randomized phase
II
PL05.06: IMPROVED QUALITY OF LIFE IN
PATIENTS UNDERGOING PLEURECTOMY AND
DECORTICATION FOR MALIGNANT PLEURAL
MESOTHELIOMA
Wickii T. Vigneswaran1, Diego Avella Patino2, Diana Kircheva2,
Sydeaka Watson2, Aliya Husain3 , Hedy Kindler2, Buerkley Rose2,
Amy Durkin2
Thoracic And Cardiovascular Surgery, Loyola University Medical
Center, Maywood, UNITED STATES OF AMERICA, 2University
of Chicago Medicine, Chicago, IL, UNITED STATES OF AMERICA, 3Department of Pathology, The University of Chicago, Chicago, IL, UNITED STATES OF AMERICA
1
Objectives: Pleurectomy and decortication (PD), a maximal cyto-reductive surgery for Malignant Pleural Mesothelioma (MPM)
improves survival in selected patients, in others the survival
benefit is modest. In a preliminary review we reported improvement in quality of life (QoL) following PD. We report our findings
in a larger cohort of patient with longer follow-up.
Methods: Patients undergoing PD were prospectively enrolled
between 2010 -2015 to determine the effects of surgery on QoL.
EORTC QLQ-C30 was utilized to assess the QOL at baseline,
and 1, 4 -5, 7- 8, and 10-11 months (m) postoperatively. Global
health, variables in functional domain and variables symptoms
domains were investigated. Sub-group analysis were also
performed for comparing preoperative performance status (PS,
0 vs 1&2), histology (epithelioid vs non-epithelioid and pathological tumor volume (PV, <600 ml vs >600mls). Survival was
summarized using Kaplan-Meier estimates; Log-rank tests were
used to compare subgroups.
Results: 114 patients were enrolled. Median age: 70 years
(range: 50-88). PS0: 35 (30.7%), PS1: 74 (64.9%), PS2: 5 (4.4%).
Epithelioid histology: 61 (53.5%), Median volume: 575ml, ranging
100-2200ml, volume<600ml: 58 (50.9%). Overall global health
worsened at the first post-operative month (p = 0.0005) returning
to baseline at 3-4 months with subsequent improvement. Non
epithelioid histology, PS 1&2 and PV > 600mls did not show
deterioration at 1 month following PD, and remained unchanged.
In functional domain, physical functioning, role functioning and
social functioning deteriorated at 1 month, cognitive function was
not altered whereas emotional functioning significantly improved.
All measures continued to improve during the follow-up. In the
symptoms domain pain, fatigue and insomnia were worse at 1
month in all groups but dyspnea was worse only in PS 0, epithelioid and in patients with tumor volume <600 ml. The overall
survival was significantly better among patients with epithelioid
histology (19.9m p<0.0001), PS 0 (19.1m, p=0.031), and tumor
volume <600ml (19.1, p<0.001).
Conclusion: Epithelioid histology, good PS, and low tumor
volume correlate with good survival in MPM. Improvement in
the QoL was observed after the first month and maintained
at late follow-up. QoL measures however were not adversely
affected by PD at any time in patients with PS 1&2, non-epithelioid histology and tumor volume >600 ml, a trend towards
improvement at late follow-up was observed. The net benefit of
PD justifies the procedure in majority of patients with MPM.
Keywords: pleural mesothelioma, pleurectomy and decortication, quality of life
iMig2016.ORG
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ABSTRACT BOOK
PL06: THE “IMMUNE WAR” ON
MESOTHELIOMA
11:15 – 12:45
PL06.05: THE IMMUNE LANDSCAPE OF HUMAN
MESOTHELIOMA TO PREDICT RESPONSE TO ANTIPD1 THERAPY
Astero Klabatsa1, Jennifer H. Yearly2, Erin Murphy2, Terri Mcclanahan2, Andrew Kossenkov3 , Daniel Sterman4 , Evan Alley5,
Leslie Litzky6 , Edmund Moon7, Steven Albelda1
Division of Pulmonary, Allergy And Critical Care Medicine,
Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, UNITED STATES OF AMERICA, 2Merck & Co., Inc,
Kenilworth, NJ, UNITED STATES OF AMERICA, 3Bioinformatics
Facility, The Wistar Institute, Philadelphia, PA, UNITED STATES
OF AMERICA, 44. pulmonary, Critical Care And Sleep Medicine,
NYU Langone Medical Center, NY, NY, UNITED STATES OF
AMERICA, 5Haematology/oncology, Perelman School of Medicine,
University of Pennsylvania, Philadelphia, PA, UNITED STATES OF
AMERICA, 6Pathology And Laboratory Medicine, Hospital of The
University of Pennsylvania, Philadelphia, AL, UNITED STATES OF
AMERICA, 7Division of Pulmonary, Allergy And Critical Care Medicine, Perelman School of Medicine, University of Pennsylvania,
Philadelphia, PA, UNITED STATES OF AMERICA
1
Objectives: With the approval of anti-PD1 and anti-CTLA4
antibody therapy for lung cancer and melanoma, there is great
interest in using these approaches in mesothelioma, and in
fact, early trials have looked encouraging. One major issue to
emerge from checkpoint inhibitory therapies is selecting those
patients most likely to respond. The biomarker currently being
tested is expression of PD-L1 on tumor biopsies, however, in
lung cancer, the value of this marker has been controversial.
Recently, a more comprehensive classification that takes into
account the presence of PD-L1 and tumor infiltrating lymphocytes (TILs) has been proposed (Teng et al, Cancer Res 2015). In
this scheme, tumors are divided into four groups: type I (TIL+/
PD-L1+), type 2 (TIL-/PD-L1-), type 3 (TIL-/PD-L1+), and type
4 (TIL+/PD-L1-). Type 1 would be predicted to have the best
responses to anti-PD1 therapy and type 2 the worst.
Methods: To study the immune landscape of mesothelioma
using this scheme, we used nanostring technology to quantify
mRNA expression levels of ~600 immune-related genes. We
also did immunohistochemical staining for PD-L1 (scored on a
6 point scale). We studied 53 malignant mesothelioma tumors
of both epithelioid (72%) and non-epithelioid (28%) histology. Results: 44% of the tumors showed high (>2 on a scale of 0-5)
PD-L1 expression. The expression of high PD-L1 was greater on
non-epithelioid tumors (67%) than on epithelioid tumors (29%).
15% (8/53) of the tumors were type 1 (TIL+/PDL1+). This group
also had the highest expression of PD1. Surprisingly, 6 of the 8
tumors in this group were non-epithelioid. 30% (16/53) tumors
were type 2 (TIL-/PD-L1-), a group that would not be expected to
respond to checkpoint therapy. 28% (15/53) tumors were type
3 (TIL-/PD-L1+) and 29% (14/53) were type 4 (TIL+/PD-L1-). T
cell infiltration was highly linked to expression of the chemokines CXCL9, 10, 11 and CCL5. Macrophage infiltration and
IFN-related genes such as IRF7, OAS2, MX1, and STAT1 seemed
to be relatively similar in each group.
Conclusion: These data show that the immune landscape of
mesothelioma is heterogeneous. About 45% of tumors seem to
have significant T cell infiltration. 15% of the tumors also have
high expression of PD1 and P-DL1 and would thus be predicted
to be highly responsive to anti-PD1 or anti-PD-L1 therapy. However, these data are only hypothesis-generating (as this scheme
has not yet been validated) and need to be paired with clinical
trial data in which the response to anti-PD1 or anti-PD-L1 therapy is determined.
Keywords: classification, nanostring, checkpoint inhibitors,
immune profile
iMig2016.ORG
11
ABSTRACT BOOK
MINI SYMPOSIUM
MS01: MARF INTERNATIONAL MESO UK
MINI SYMPOSIUM
MONDAY, MAY 2, 2016
14:15 – 15:45
MS01.04: PROGRAMME FOR NURSES TO IMPROVE
COMMUNICATION SKILLS IN THE CARE OF
PEOPLE WITH MESOTHELIOMA IN JAPAN
Yasuko S. Nagamatsu1, Helen Clayson2, Tsuyoshi Matsuda3
programme either very much agree or agree regarding overall
satisfaction with the programme, comprehension and relevance. Comments from participants: Apart from the positive
feed back of the program, 6 requested further program to
improve communication skills including role-play. They reported that high level communication skills were important in 2
particular areas: 1) when, as routine in Japan, the diagnosis is
disclosed firstly to relatives who then do not want this information to be given to the patient, and 2) when relatives demand
inappropriate radical treatments when the patient has endstage disease.
Conclusion: This programme to improve communication skills
for nurses caring for people affected by mesothelioma was
highly rated by participants. It offers a model that could apply
to similar settings.
Keywords: education, nursing, communication skill, mesothelioma
Nursing, St. Luke’s International University, Tokyo, JAPAN, 2Centre for the Social History of Health and Healthcare, Glasgow,
UNITED KINGDOM, 3Kobe University, Kobe, JAPAN
1
Objectives: Malignant Pleural Mesothelioma (MPM) causes
1400 deaths/year in Japan. Following educational programmes
about mesothelioma (2 days in 2012 and 1 day in 2013) nurses
reported ongoing difficulties in talking with people suffering
from mesothelioma, and their relatives. To address this a 1 day
communications skills programme was developed in 2014. It
included 1) clinical and psychological aspects of MPM, 2) interactive lecture on communication skills including ‘breaking bad
news’ ,3) videos illustrating good and bad communication skills
4) narratives of bereaved relatives and 5) group discussion. The
aim of this study is to report and evaluate the programme.
Methods: Recruitment: Letters of invitation to nurses were
sent nationwide to heads or nursing directors of health care
facilities. Ethical considerations: Ethical principles were
followed: avoiding harm, voluntary participation, anonymity
and protection of privacy and personal information. Participants were informed that this program included evaluation.
The purpose, procedure and confidentiality of the study were
explained verbally at the outset of the course and in written
format. Participants were informed that nonparticipation would
not disadvantage them. Data were collected from those who
wished to participate and who completed the informed consent
form. 1. Participants’ satisfaction form: Participants provided feedback regarding the programme by responding to the
following four items: 1) satisfaction with the overall program; 2)
comprehension of the content and　3) relevance of the content.
The programme was evaluated using a 5-point Likert scale
(5=very much agree to 1=never agree). Higher scores indicated
more positive feedback for the program. 2. Comment form: This
was the first educational program about communication
skills in MPM therefore, it was important to capture as much
feedback as possible. Participants were encouraged to provide
written comments about their experience of the program in an
open-ended format.
Results: Participants : Twenty seven nurses had worked for
for 1-35 years (mean = 15.2). Current posts were: respiratory
department (8), palliative care (6), OPD (4), home visiting care
(2), hospice (2) and health centre (1). Five had no experience
of MPM. Participants’ satisfaction: All participants rated the
MS01.05: MESOTHELIOMA PATIENTS’ AND
CARERS’ CONCERNS ABOUT THEIR DIAGNOSIS,
TREATMENT, AND CARE
Richard Stephens1, Helen Clayson2, Heather Foot3 , Kate Hill4 ,
Ian Jarrold5, Katherine Cowan6 , Caroline Whiting6
ex MRC Clinical Trials Unit, London, UNITED KINGDOM, 2Centre for Social History of Health and Healthcare, University of
Strathclyde and Glasgow Caledonian University, Glasgow,
UNITED KINGDOM, 3(Bereaved Carer), Matlock, UNITED
KINGDOM, 4Leeds Institute of Health Sciences, Leeds, UNITED
KINGDOM, 5British Lung Foundation, London, UNITED KINGDOM, 6James Lind Alliance, Southampton, UNITED KINGDOM
1
Objectives: In July 2013 the UK Parliament approved measures to increase awareness of, and support research into, mesothelioma. These included a James Lind Alliance (JLA) Priority
Setting Partnership (PSP), funded by the National Institute for
Health Research (NIHR), which brought together patients and
carers, relevant health professionals, and patient support organizations, to identify and prioritize specific interventions that
could be tested in a clinical trial setting. The project involved
an initial survey to identify areas where research was needed.
From the 453 responses 50 unanswered research questions
were generated. A second survey prioritized these questions,
and the top 30 were taken to a workshop where a final prioritization was agreed. These results have been published1. However, the initial responses also identified numerous issues that fell
outwith the remit of the project. The Steering Group of the JLA
PSP nonetheless recognized the importance of these issues,
and felt that their implementation would significantly improve
the experience of mesothelioma patients (and their carers) who
have to cope with this devastating disease.
Methods: All the responses to the initial survey were reviewed,
and those not eligible for inclusion in the original NIHR remit
(i.e. a question involving an intervention that could be tested)
were grouped under 4 themes proposed by an existing framework for quality of care2.
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ABSTRACT BOOK
Results: Patients and carers raised concerns regarding all
4 aspects of care: 1. The medical and technical competence
of health professionals (lack of compassion, knowledge and
competence) – ‘‘You have mesothelioma and it will kill you
within 2 years’ .. words used by my husband’s consultant without
preamble’ 2. The physical and technical conditions of the care
organisations (inefficient uncoordinated care, poor communication, and lengthy delays) – ‘The inconsistencies are unacceptable. Every mesothelioma patients deserves the best care and
treatment. This shouldn’t depend on which hospital you attend’ 3.
The degree of identity orientation in attitudes and actions of
caregivers (technical complexity of the disease, and supporting
patients and carers) – ‘One of my daughters (a nurse) sighed
and summed it all up thus ‘we still don’t do dying very well’’ 4.
The socio-cultural atmosphere of the care organization – ‘There
was a distinct lack of support for us as a family when Dad was
diagnosed’
Conclusion: Patients are still being diagnosed with mesothelioma in increasing numbers across the UK, and consequently
new therapies and improvements in care are required for
people coping with this devastating disease. This JLA PSP survey highlighted many concerns with diagnosis, treatment and
care, all of which contribute to patients (and carers) feelings
of abandonment, loss, bewilderment and frustration. Many of
these issues have been reported previously but remain commonly experienced and unresolved3 . However, addressing these
concerns, could, and should, improve the current distressing
experience for patients, and help support their families and
carers. 1 Stephens RJ et al. Lung Ca 2015, 89, 175-80 2 Wilde B et
al. Scand J Caring Sci 1994, 8, 39-48 3 Clayson H et al. Hematol
Oncol Clin N Am 2005, 19, 1175-90
Keywords: James Lind Alliance, Quality of Care, Patients and
Carers
MS01.06: ASBESTOS-RELATED DISEASE SUPPORT
GROUPS: A SURVEY OF THEIR ORGANISATIONAL
STRUCTURES AND ACTIVITIES
Helen Clayson1, Kate M. Hill2
Centre For The Social History Of Health And Healthcare, Glasgow
Caledonian University, Glasgow, UNITED KINGDOM, 2Leeds Institute of Helath Sciences, The University of Leeds, LEEDS, UNITED
KINGDOM
1
day organisation and operational activity and (2) to develop a
survey instrument that could be used to monitor and support
group development and impact in the future.
Methods: A survey form, developed by the authors in collaboration with the ASVGs Forum UK, was distributed by email in
December 2015 to 14 Forum support groups. Nine responses
(64%) have been received to date. The survey remains open
until 31 January 2016 and it is predicted that the response rate
will rise when reminders are sent out in mid-January.
Results: The 9 groups were located in areas traditionally associated with substantial industrial use of asbestos. They were
set up between 5 and 20 years ago (5-10 years: n=3, >10 years:
n=5, 20 years: n=1) by independent health and safety organisations (n=2), trade unions (n=2), occupational health organisation (n=1), bereaved relatives (n=3) and in one case a doctor, in
response to unmet needs of people with ARDs, particularly mesothelioma. All the groups provided expert advice and practical
assistance with state benefits, usually in people’s homes and by
telephone, sometimes on group premises and occasionally by
email. All supplied details of expert solicitors. Four held regular
meetings for people with ARDs at which benefits advisors, specialist nurses and sometimes solicitors were present, 3 offered
medical information and 4 offered bereavement support. Six
held annual educational meetings open to patients, healthcare
professionals and lawyers. Three groups expressed concerns
around sustainability due to insecure funding In 2015 these 9
groups supported 2460 people affected by ARDs including 963
with mesothelioma, 242 with asbestos-related lung cancer,
451 with asbestosis, 1 with laryngeal cancer and 359 bereaved
relatives.
Conclusion: Forum AVSGs provide essential psychosocial
and practical support for a large number of people affected by
ARDs, particularly in offering expert assistance with complex
financial and legal matters that are a unique and additional
burden in mesothelioma and asbestos-related lung cancer.
Some groups also provide emotional and practical support for
relatives, including in bereavement. The results reveal that a
disproportionately small number of people with asbestos-related lung cancer access the groups Activity data demonstrate
initiatives, eg bereavement support and education, that could
benefit others affected by ARDs and those who support them.
Keywords: psychosocial support, asbestos victim support
groups, State benefit and civil compensation claims, mesothelioma
Objectives: Mesothelioma is a devastating disease that has
a high emotional impact on patients, their families and carers.
Patient organisations play an important role in the supportive
care of patients with asbestos-related diseases (ARDs), especially those with mesothelioma. The psychosocial sequelae and
complex benefit and compensation claims associated with mesothelioma have resulted in a need that is not met by standard
health and social care services. The Asbestos Victims Support
Groups (AVSGs) were founded in response to this unmet need.
Members of the AVSGs Forum UK adhere to a set of principles
that includes specifying the nature of their relationships with
lawyers. The aim of this study was (1) to provide a comprehensive picture of the way Forum AVSGs are set-up, their day-toiMig2016.ORG
13
ABSTRACT BOOK
MS02:CELL DEATH MECHANISMS
MONDAY, MAY 2, 2016
14:15 – 15:45
MS02.02: SIGNALLING PATHWAYS INVOLVED IN
UPREGULATION OF MCL1 IN MM; METABOLIC
REPROGRAMMING PROVIDES A NOVEL
APPROACH TO SENSITISE MM
novel approach to selectively sensitize malignant mesothelioma
cells to targeted agents such as the BH3-mimetic, ABT-737.
Keywords: Metabolism, Signal transduction, Cell death,
Pre-clinical models
MS02.03: MONOAMINE OXYDASE A AS A
POTENTIAL NEW TARGET TO TREAT MALIGNANT
PLEURAL MESOTHELIOMA
Xiao-Ming Sun1, Gareth J. Miles1, Ian R. Powley1, Sara Galavotti1, Tatyana Chernova1, Stefano Grosso1, Jonathan Bennett2,
Apostolos Nakas2, Martin Bushell1, Anne E. Willis1, Kelvin Cain1,
Marion Macfarlane1
David Roulois, Sophie Deshayes, Marc Grégoire, Christophe
Blanquart
MRC Toxicology Unit, Leicester, UNITED KINGDOM, 2Glenfield
Hospital, UHL NHS Trust, Leicester, UNITED KINGDOM
Objectives: Malignant Mesothelioma (MM) is an asbestos-related cancer and is currently resistant to the entire chemotherapeutic regime. In most tumour cells, the Warburg effect (aerobic
glycolysis) not only provides building blocks for the synthesis of
macromolecules, but also provides tumour cells with a survival
advantage via constitutive activation of pro-survival signalling
pathways. Understanding the molecular basis of this aerobic
glycolysis-mediated pro-survival signalling in MM could provide
important new insights to help tackle the widespread resistance
of MM to the current regime of cancer chemotherapeutics.
Objectives: Malignant pleural mesothelioma (MPM) is a rare
and aggressive cancer related to asbestos exposure. Therapeutic actions are limited mainly due to late diagnosis and resistance of mesothelioma cells to treatments. Given the failure
of the current therapies to improve significantly mesothelioma
outcome, new strategies need to be developed. Using a transcriptomic approach, we identified monoamine oxidases (MAO)
as interesting targets in mesothelioma. MAO are implicated in
monoamines catabolism such as serotonin. Two isoforms of
MAO have been described: MAO-A and MAO-B. In this work, we
assessed the inhibition of these enzymes, alone or in combination with cisplatin, as a new therapeutic option to treat MPM.
Methods: A panel of primary malignant mesothelioma (MM)
cell lines, freshly-derived from patient tumours (Chernova, Sun
et al, Cell Death Differ., in press) were used to assess drug sensitivity and drug resistant mechanisms. MM cells were treated
with 2-deoxyglucose (2DG) to inhibit glycolysis. Apoptotic cell
death was assayed by FACS, using ΔΨm or PS externalisation.
Western blotting and siRNA knockdown of key proteins was carried out according to standard protocols. Oxidative phosphorylation (OCR) and glycolysis (ECAR) were measured in live cells
using a Seahorse BioScience XF Analyzer. Freshly-resected
3D tumour explants, cultured ex-vivo, were used to assess the
rational combination of potential therapeutic reagents in MM.
Methods: This study was performed using our biocollection of
MPM cell lines established from pleural effusions of mesothelioma patients. The mRNA expressions were measured using
real-time PCR. In a first step, clorgyline was used to inhibit
MAO-A and pargyline was used to inhibit MAO-B. Evaluation
of toxicity of the treatments was performed by measuring cell
viability. Apoptosis was also studied by labelling cells with
annexin-V-FITC and propidium iodide, and by measuring mitochondrial potential. In a second step, clorgyline was combined
with cisplatin, with primary mesothelial cells used as controls.
Finally, the efficacy of the molecules was assessed on spheroids of MPM cells.
Results: In this study, we have uncovered a link between
aerobic glycolysis and signal transduction that appears to be
responsible for increased levels of the anti-apoptotic protein,
MCL-1 in MM. Signal transduction analysis revealed that
2DG-induced downregulation of MCL-1 is mediated by inhibition
of Stat3-mediated MCL-1 transcription. 2DG also induced a
concomitant activation of AKT, by which total MCL-1 degradation was inhibited, possibly through inactivation of GSK-3β. In
combination with a specific AKT inhibitor, AZD-5363, complete
clonogenic cell death was achieved in the presence of 2DG and
the Bcl-2/Bcl-xL inhibitor/BH3-mimetic, ABT-737. Importantly,
in MM patient freshly-resected 3D tumour explants, which
retain the tumour microenvironment, 2DG/AZD-5363 -mediated
downregulation of MCL-1 correlated with induction of tumour
cell death.
Results: Transcriptomic analysis revealed an increase of the
MAO-A/MAO-B expression ratio in MPM cells compared with
primary mesothelial cells. The MAO-A inhibitor clorgyline
decreased MPM cell viability, whereas the MAO-B inhibitor
Pargyline had no effect. The toxicity of clorgyline was related to
apoptosis induction. An additive toxicity was observed on MPM
cells when clorgyline was combined with cisplatin compared
with drugs used alone. The toxicity of the combination was higher on MPM cells than on primary mesothelial cells. Finally, a
potentiation of apoptosis induction was observed on spheroids
of MPM cells treated with clorgyline and cisplatin, compared
with drugs used alone.
1
Conclusion: Down regulation of MCL-1 levels, by inhibition
of glycolysis with 2-DG or via 2-DG in combination with other
well-tolerated and clinically approved therapeutics, provides a
Centre de Recherche contre le Cancer Nantes et Angers, University of
Nantes, CNRS UMR 6299, Inserm U892, Nantes cedex, FRANCE
Conclusion: All these data highlight MAO-A as an interesting
new target to treat mesothelioma. Additional investigations on
preclinical model are required to validate this hypothesis.
Keywords: mesothelioma, Monoamine oxydase, cisplatin,
chemotherapy
iMig2016.ORG
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ABSTRACT BOOK
MS02.04: EGFR TYROSINE KINASE INHIBITORS
OVERCOME RESISTANCE TO CHEMOTHERAPY IN
MALIGNANT PLEURAL MESOTHELIOMA
Bernard Staumont1, Chrisostome Costa1, Fabian Vandermeers1,
Sathya Neelature Sriramareddy1, Ludovic Dhont1, Céline Mascaux2, Arnaud Scherpereel3 , Bernard Duysinx4 , Renaud Louis4 ,
Philippe Delvenne5, Pascale Hubert5, Luc Willems1
Molecular And Cellular Epigenetics, Interdisciplinary Cluster
for Applied Genoproteomics (GIGA) of University of Liège, Liège,
BELGIUM, 2Faculty Of Pharmacy, UMRS911, University of Marseille, Marseilles, FRANCE, 3Calmette Hospital, CHRU of Lille and
University of Lille II, Lille, FRANCE,4Pneumology, CHU of Liège,
Liège, BELGIUM, 5Laboratory of Experimental Pathology, Interdisciplinary Cluster for Applied Genoproteomics (GIGA) of University
of Liège, Liège, BELGIUM
1
Objectives: Malignant pleural mesothelioma (MPM) is a cancer of the pleura mainly caused by exposure to asbestos fibers.
Current treatments are unsatisfactory due to intrinsic chemoresistance of the tumor. We hypothesized that chemoresistance
was due to epigenetic errors and evaluated the ability of HDAC
inhibitors to improve treatment efficacy. We previously showed
that valproic acid (VPA) improves the first line regimen of MPM
both in vitro and in vivo (Vandermeers et al, 2009, Clinical Cancer Research 15: 2818). A clinical trial also demonstrated that
VPA in combination with doxorubicin increases the response
rate of second line patients (Scherpereel et al, 2011, European
Respiratory Journal 37:129).
Methods: Transcriptomic profiling was performed by microarray analyses (Agilent). Gene expression was validated by quantitative RT-qPCR. Modulation of TGFα expression was performed
by shRNA interference and transfection of a cDNA vector. Onset
of apoptosis was assessed with the Annexin V assay.
Results: To evaluate the mechanisms associated with the
response to chemotherapy, we compared two types of MPM cell
lines (M14K and H28) characterized by a difference in sensitivity to doxorubicin+VPA. Microarray analyses and bioinformatic
modeling of gene expression profiles revealed the most relevant
candidate genes associated with sensitivity or resistance to this
regimen. Among these, TGFa expression was associated with
resistance to doxorubicin + VPA in a series of MPM cell lines.
Silencing of TGFα by RNA interference in H28 cells correlated
with a significant increase in apoptosis. On the other hand,
overexpression of TGFα desensitized M14K cells to doxorubicin+VPA -induced apoptosis. Since TGFα interacts with the
EGF receptor, we evaluated pharmacological inhibition using
EGFR tyrosine kinase inhibitors (erlotinib and gefitinib) and the
dual HDAC/EGFR inhibitor CUDC-101. As predicted, these TKI
inhibitors improved efficacy of doxorubicin+VPA.
MS02.05: TISSUE TRANSGLUTAMINASE (TG2):
A POTENTIAL NOVEL TARGET FOR HUMAN
TREATMENT
Sara Zonca1, Giulia Pinton1, Maria Felicia Soluri2, Szilvia Bakó2,
Daniele Sblattero3 , Laura Moro1
1
Pharmaceutical Sciences, University of Piemonte Orientale,
Novara, ITALY, 2Health Sciences, University of Piemonte Orientale, Novara, ITALY, 3Life Sciences, University of Trieste, Trieste,
ITALY
Objectives: Characterize the expression and function of Tissue
Transglutaminase (TG2) in human malignant pleural mesothelioma (MPM) cell models.
Methods: TG2 isoforms expression has been evaluated by Real
time-PCR and Western Blot analyses in normal mesothelium
and MPM derived cell lines grown under normoxic and hypoxic
conditions.
Results: We demonstrate that cells derived from biphasic MPM
express higher total and surface TG2 levels than cells derived
from epithelioid MPM and normal mesothelium. We firstly
evidence that the full length TG2-v1 is the highest expressed
TG2 isoform both in mesothelial and MPM cells; instead, only
low levels of the other described variants (v2, 3, 4 and 5) are expressed. We observe a significant induction of TG2 when MPM
cells are grown 48 hours as monolayer in hypoxia or packed in
spheroids, where the presence of a hypoxic core is demonstrated. We describe the HIF2 dependent hypoxic induction of TG2.
Importantly, while the silencing of TG2 in MPM cells in normoxia causes only a modest reduction in cell viability, its silencing
in hypoxia causes a reduction by more than 80%. Furthermore,
TG2 silencing results in a marked decrease in MPM spheroids
volume.
Conclusion: MPM is a tumor with significant areas of hypoxia;
understanding of the expression and function of TG2 in the
adaptation to the hypoxic environment may provide useful
information for novel promising therapeutic option for MPM
treatment.
Keywords: tissue transglutaminase, hypoxia, Malignant pleural
mesothelioma, cell viability
Conclusion: Our data demonstrate that TGFα is involved in
resistance of MPM to chemotherapy and that TKI inhibitors
overcome resistance to second line regimen.
Keywords: valproic acid, EGFR TKI, TGFalpha, chemoresistance
iMig2016.ORG
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ABSTRACT BOOK
MS02.06: AUTOPHAGY INHIBITION SENSITIZES
PRIMARY MALIGNANT MESOTHELIOMA TO A
DUAL PI3K/MTOR INHIBITOR
Sara Galavotti1, Tatyana Chernova1, Xiao-Ming Sun1, Ian R.
Powley1, Gareth J. Miles1, David Dinsdale1, Jonathan Bennett2,
Apostolos Nakas2, Anne E. Willis1, Kelvin Cain1, Marion Macfarlane1
Hospital, University Hospitals of Leicester NHS Trust, Leicester,
UNITED KINGDOM
1
Objectives: Malignant mesothelioma (MM) is a rare, aggressive tumour arising from the mesothelial lining cells of the
pleura and commonly associated with asbestos exposure. In
MM the phosphatidylinositide-3-kinase/mammalian target of
rapamycin (PI3K/mTOR) signalling pathway is constitutively
active, promoting tumour growth. Inhibition of mTOR can lead
to induction of autophagy and increased resistance to mTOR
inhibitors, while inhibition of PI3K alone is insufficient to induce
apoptosis. Importantly, a single agent dual inhibitor of PI3K/
mTOR is now feasible due to the development of pan PI3K/
mTOR inhibitors, several of which are currently undergoing clinical trials. Autophagy is an evolutionarily conserved catabolic
pathway involved in the degradation of cytoplasmic macromolecules and organelles to satisfy cellular energetic and nutritional needs. Of note, autophagy has been linked to increased
resistance to drug treatments, including mTOR inhibitors.
Methods: We used a panel of freshly-derived primary mesothelioma cell lines (Chernova, Sun et al, Cell Death Differ., in press)
and surgically-resected 3D tumour-explants cultured ex-vivo which retain the tumour microenvironment. Induction of autophagy was confirmed by electron microscopy and autophagic
flux measured via western blot (LC3-II/LC3-I) and immunofluorescence/immunohistochemistry. Metabolic profiling was
performed using a Seahorse Extracellular Flux Analyser. Cell
death was quantified by Annexin V/PI staining or in the case of
3D tumour explants by immunostaining for cleaved PARP.
Results: In primary mesothelioma cell lines, we show that dual
inhibition of PI3K/mTOR induces substantial autophagy and
promotes cell survival. However when autophagy is blocked
we observe an increase in cell death which is caspase-independent. Since autophagy is a metabolic process, we explored
whether the dual PI3K/mTOR inhibitor had any effect on cellular
bioenergetics. Cells treated with the dual inhibitor indeed
exhibit reduced levels of ATP and mitochondrial oxidative phosphorylation, confirming the loss of cellular energy homeostasis.
Importantly, in 3D tumour-explants freshly resected from MM
patients inhibition of autophagy significantly enhances sensitivity to the dual PI3K/mTOR inhibitor.
Conclusion: Together, these data suggest a role for autophagy
in the modulation of survival in MM and provides a rationale for
targeting autophagy in MM patients. In addition, our findings
reveal that inhibition of the PI3K/mTOR pathway compromises cellular energy homeostasis unveiling potential metabolic
vulnerabilities in MM that could be exploited using combination
therapies.
MS02.07: AUTOPHAGY CORRELATES WITH
PATIENT OUTCOME IN MESOTHELIOMA
Carlo Follo1, Dario Barbone1, William G. Richards2, Raphael
Bueno2, Courtney Broaddus1
Medicine/pulmonary, San Francisco General Hospital, University
of California San Francisco, San Francisco, CA, UNITED STATES
OF AMERICA, 2Brigham and Women’s Hospital, Boston, MA,
1
Objectives: Autophagy, a degradation process that eliminates
dysfunctional proteins and organelles and thereby provides
energy and amino acids, may play an important role in cancer,
although its actual role is still unclear. Understanding the role
of autophagy in cancer has been limited by the inability to
measure this dynamic process in formalin-fixed tissue. We considered that three-dimensional models including ex vivo tumor,
such as we have developed for our research in mesothelioma,
would provide valuable insights. Using these models, in which
we could inhibit lysosomal proteases to measure the autophagic degradation activity, or autophagic flux, we sought a marker
of autophagy that would be valid in formalin-fixed tumor and be
used to assess the role of autophagy in patient outcome.
Methods: Autophagy was studied in mesothelioma cell lines,
as two-dimensional (2D) monolayers and three-dimensional (3D) multicellular spheroids, and in tumor from 25 chemonaive patients, both as ex vivo 3D tumor fragment spheroids (TFS)
and as formalin-fixed tissue. Autophagy was evaluated as autophagic flux by detection of the accumulation of a key protein
in the autophagic process (LC3) after inhibition of lysosomal
proteases. Autophagy was also evaluated as autophagy initiation by detection of ATG13 puncta. ATG13 is a protein involved
in the early phase of autophagy, the initiation phase. When
autophagy is activated, ATG13 accumulates in structures at the
forming autophagic vesicles that can be detected as puncta by
immunofluorescence.
Results: We found that autophagic flux in 3D, but not in 2D,
correlated with ATG13 positivity. In each TFS, ATG13 positivity
was similar to that of the original tumor from which the TFS was
generated. When tested in tissue microarrays of 109 chemonaive patients, higher ATG13 positivity correlated with better outcome and a longer time to progression and provided prognostic
information independent of known prognostic factors.
Conclusion: Our results show that ATG13 is a static marker
of the autophagic flux in 3D models of mesothelioma and may
also reflect autophagy levels in formalin-fixed tumor. If confirmed, this marker would represent a novel prognostic factor
for mesothelioma, supporting the notion that autophagy plays
an important role in this cancer. In conclusion, we have used 3D
models of mesothelioma to identify a marker of autophagy that
in turn has prognostic value in a group of patients with mesothelioma. Our hope is to use these models to explore the role
of autophagy in this tumor.Research support from the Simmons
Mesothelioma Foundation.
Keywords: Autophagy, three-dimensional models, outcome,
ex vivo
Keywords: PI3K/mTOR inhibition, Metabolism, Autophagy, Cell
Death
iMig2016.ORG
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ABSTRACT BOOK
MS03: IMAGING AND ENDPOINT EVALUATION
MONDAY, MAY 2, 2016
14:15 – 15:45
MS03.01: INDOCYANINE GREEN AND
INTRAOPERATIVE IMAGING DETECTS RESIDUAL
DISEASE FOLLOWING RESECTION OF MALIGNANT
Jane Keating, Jarrod Predina, Ollin Venegas, Sarah Nims,
John Kucharczuk, Sunil Singhal
Surgery, University of Pennsylvania, Philadelphia, PA, UNITED
STATES OF AMERICA
Objectives: Patients with epithelioid malignant mesothelioma limited to the hemithorax benefit from an approach
that includes surgery with the goal of macroscopic complete
resection. At the conclusion of surgery, it can be challenging to
discriminate residual disease from scar and normal tissue. We
propose near-infrared (NIR) intraoperative molecular imaging
with indocyanine green (ICG) for the detection of mesothelioma
tumor deposits for more complete macroscopic resection.
Methods: Eight patients with biopsy proven malignant pleural
mesothelioma were enrolled in a pilot clinical trial. All patients
underwent 5 mg/kg of intravenous ICG injection. The following
day, a NIR imaging device was used to detect fluorescence
intraoperatively. After what was believed to be complete tumor
excision, the wound bed was reimaged for residual fluorescence
indicative of retained tumor, and additional tissue was resected
when feasible. Specimens were sent for pathological correlation.
Results: All patients underwent ICG injection with no evidence
of drug toxicity. NIR fluorescence localized to mesothelioma in
all cases intraoperatively and fluorescence was confirmed on
the back table. The mean in vivo NIR tumor-to-background ratio
was 3.2 (IQR 2.9-3.4). Residual disease was discovered upon
wound bed imaging in all 8 patients. The number of resected
specimens following wound bed imaging ranged from 1 to 4
(average 1.8). Disease was typically discovered in difficult to
reach places, including the costophrenic sulcus and directly
beneath or adjacent to the thoracotomy incision. The mean
tumor-to-background ratio of the resected residual tumor
deposits was 2.8 (IQR 2.6- 3.1). Additionally, these specimens
ranged in size from 0.3 mm to 2.2 cm (mean 0.9 cm). In all
cases, the additionally resected fluorescent tissue was malignant mesothelioma on pathology. Conclusion: NIR intraoperative molecular imaging using ICG
localizes to malignant pleural mesothelioma and aids in detection of residual disease for improved resection. A larger clinical
trial is needed to investigate the impact of NIR intraoperative
imaging on patient survival.
Keywords: Near-Infrared, Intraoperative Imaging, Indocyanine
Green
MS03.02: THE VALUE OF DELAYED PHASE
ENHANCEMENT FOR MAGNETIC RESONANCE
IMAGING OF MALIGNANT PLEURAL
MESOTHELIOMA
Sharyn I. Katz1, Akash Patel2, Ian B. Berger3 , Urooj Khalid3 ,
Drew A. Torigian2, Charles B. Simone4 , Andrew Haas3 ,
Evan Alley5, Sunil Singhal6 , Keith A. Cengel4
Radiology, University of Pennsylvania Perelman School of
Medicine, Philadelphia, UNITED STATES OF AMERICA, 2Radiology, University of Pennsylvania Perelman School of Medicine,
Philadelphia, PA, UNITED STATES OF AMERICA, 3University of
Pennsylvania Perelman School of Medicine, Philadelphia, PA,
UNITED STATES OF AMERICA, 4Radiation Oncology, University
of Pennsylvania, Philadelphia, PA, UNITED STATES OF AMERICA, 5Haematology/Oncology, Perelman School of Medicine,
AMERICA, 6Surgery, University of Pennsylvania, Philadelphia, PA,
1
Objectives: Radiologic staging of malignant pleural mesothelioma (MPM) on cross-sectional imaging can be challenging
when evaluating for the presence of subtle local invasion.
Since accurate staging is vital to inform treatment decisions,
techniques that optimize pleural imaging are critical. Here we
characterize the kinetics of MPM enhancement on magnetic
resonance imaging (MRI).
Methods: All MPM patients with intravenous (IV) contrast
enhanced staging thoracic MRI between 2008-2014 at our
institution were retrospectively selected for image analysis.
Patients with maximum pleural tumor thickness <1 cm were
excluded. Quantitative measurements of tumor signal were
obtained on pre-contrast and post-contrast phases where MRI
iMig2016.ORG
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ABSTRACT BOOK
acquisition parameters were fixed. Using best-fit model curves,
predicted maximum values were determined using a simulation
of predicted values. Additionally, a qualitative assessment of
tumor enhancement kinetics was performed. Two board-certified thoracic radiologists, blinded to the quantitative imaging
data, assessed de-identified side-by-side sets of post-contrast
images representing every time point iteration and chose the
most conspicuous tumor image. Statistical analysis assessed
for correlation between qualitative lesion conspicuity and quantitative tumor enhancement.
Results: Of the 23 MPM patients who had undergone staging
MRI, 10 patients met the exclusion criteria. Tumor enhancement
kinetics of these patients are displayed (Figure 1) as maximal
signal intensity as a function of time. Peak tumor enhancement
was at 280 seconds(s) following IV contrast administration (Figure 2). At 280s, 70%, 70% and 60% of patients are projected to
have reached >80%, >85%, and >90% of respective peak projected signal intensities. There was a correlation between degree of tumor enhancement and subjective lesion conspicuity.
Conclusion: Optimal MPM contrast enhancement on MRI
occurs at a time later than is typically imaged on routine clinical
imaging. The impact of delayed phase enhancement on radiologic MPM staging accuracy and therapy response assessment
warrants further study.
Keywords: mesothelioma, enhancement, kinetics, Imaging
MS03.03: OPTIMISATION OF THE METHODS
FOR EARLY CONTRAST ENHANCEMENT (ECE)MAGNETIC RESONANCE IMAGING IN PATIENTS
WITH MESOTHELIOMA
Selina Tsim1, Catherine A. Humphreys2, David B. Stobo3 ,
Gordon W. Cowell3 , Rosemary Woodward4 , John E. Foster4 ,
Craig Dick2, Kevin Blyth1
Respiratory Medicine, Queen Elizabeth University Hospital,
Glasgow, UNITED KINGDOM, 2Pathology, Queen Elizabeth University Hospital, Glasgow, UNITED KINGDOM, 3Radiology, Queen
Elizabeth University Hospital, Glasgow, UNITED KINGDOM, 4Clinical Research Imaging Facility, Queen Elizabeth University Hospital, Glasgow, UNITED KINGDOM
1
Objectives: We have previously reported the preliminary
diagnostic performance of a novel perfusion-based Magnetic
Resonance Imaging (MRI) biomarker of pleural malignancy (PM)
– Early Contrast Enhancement (ECE) (Thorax 2015;70:Suppl 3,
A16, doi:10.1136/thoraxjnl-2015-207770.27). Here we describe
a further analysis, which aims to resolve the potential confounding effect of signal intensity (SI) measurements from interspersed areas of benign pleural disease in patients with PM.
Methods: For measurement of ECE, T1-weighted
3D-spoiled-gradient-echo MRI sequences are acquired at
baseline, 40 seconds, 80 seconds and 4.5, 9 and 13.5 minutes
after intravenous Gadobutrol contrast. SI is measured in up to
15 regions of interest (ROI) on areas of representative parietal
pleural disease. ECE is defined objectively as an early peak in
SI (≤4.5 minutes). A patient is classified as Malignant if ECE is
demonstrated in at least one ROI, even if all others exhibit no
ECE. 18 patients had ECE assessed and subsequent histological
sampling. ROI SI gradient (ROISIG) can also be calculated, as
peak SI - baseline SI divided by time, allowing Receiver Operating Characteristic (ROC) curves to be plotted, summarising
discriminate performance across all ROIs. This approach would
allow different cut-points to be defined for tailored diagnostic
performance in subsequent studies (e.g. higher specificity for
screening asbestos-exposed individuals) but does not account
for heterogeneous tumour deposition, which is typical of MPM.
We hypothesised that the discriminant performance of ROISIG
would be improved by excluding ECE-negative ROI, if these were
areas of interspersed benign disease. To test this, we plotted a
ROC curve incorporating all ROISIG data and compared this to
one incorporating only data from ECE-positive ROI in patients
with PM. In both analyses all ROI were included in patients with
benign disease.
Results: Mean patient age was 73 (± 8) years. 12/18 had
pleural thickening ≤10mm. ECE was present in 10/11 patients
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ABSTRACT BOOK
with PM (MPM (n=10); lung cancer (n=1)) and absent in 6/7
patients with benign disease (BAPE (n=4), fibrothorax (n=2), TB
(n=1)). As previously reported, ECE demonstrated sensitivity of
91%, specificity 86%, negative predictive value 86%, positive
predictive value 91% and Inter-observer agreement 0.766. ROC
curves are shown in Figure 1.
Conclusion: In a previously presented pilot study we have
shown that ECE can be assessed in patients with minimal pleural thickening, with encouraging preliminary diagnostic results.
These additional analyses suggest that exclusion of ECE-negative ROI improves the discriminant performance of ROISIG,
probably because these areas represent interspersed benign
disease, and may enhance the method.
Keywords: Biomarkers, Imaging
iMig2016.ORG
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ABSTRACT BOOK
MS03.04: HISTOGRAM ANALYSIS OF DW-MRI
DATA DURING EARLY CHEMOTHERAPY RESPONSE
PREDICTS OUTCOME OF INOPERABLE MPM
PATIENTS
Johan Coolen1, Frederik Dekeyzer1, Philippe Nafteux1, Adriana
Dubbeldam1, Walter De Wever1, Angela Botticella2, Lieven
Depypere3 , Hans Van Veer3 , Herbert Decaluwé3 , Eric Verbeken4 ,
Stephanie Peeters2, Christophe Dooms5, Johan Vansteenkiste5,
Willy Coosemans3 , Paul De Leyn3 , Dirk Van Raemdonck3 , Kristiaan Nackaerts5, Vincent Vandecaveye1, Johny Verschakelen1
Radiology, University Hospitals of Leuven, LEUVEN, BELGIUM, 2Radiotherapy, University Hospitals of Leuven, LEUVEN,
BELGIUM, 3Thoracic Surgery, University Hospitals of Leuven,
LEUVEN, BELGIUM, 4Pathology, University Hospitals of Leuven,
LEUVEN, BELGIUM, 5Pneumology, University Hospitals of Leuven,
LEUVEN, BELGIUM
1
Objectives: Patients with unresectable malignant pleural mesothelioma (MPM) are most commonly treated with palliative
chemotherapy (PCT), while treatment efficacy is radiologically
monitored using modified RECIST criteria. However, anatomy-based assessments have limitations, one of the reasons
why determination of progression free survival (PFS) is often
difficult. Even multiparametric MR imaging (mpMRI) parameters can be insufficient for differentiating long-term and shortterm surviving patients, probably due to the large heterogeneity
of disease phenotypes, which deeply influences response to
therapy and imaging evaluation. In this study we examined the
diffusion-weighted MR imaging (DWI) values evaluating five
histogram parameters (volume[V], mean[ME], standard deviation[SD], skewness[SK], and kurtosis[KU]).
Methods: Fifteen patients with inoperable MPM were selected for systemic PCT (cisplatin-pemetrexed). MR examinations
(including DWI with 6 b-values) were performed at baseline[BA]
and after one month, just before the second chemotherapy session [DU]. ADC histograms were made for the ADCavg (calculated from all 6 b-values) and the ADClow (calculated from the
first 3 b-values ranging from 0 to 100 s/mm²), and first order
histogram statistics (V, ME, SD, SK, and KU) were checked for
differences between long-term and short-term PFS (cut-off: 170
days) and overall survival (OS, cut-off: 440 days). Mann-Whitney U tests were used to check for differences.
Results: When using baseline parameters for differentiating
between long- and short-term OS, ADClow[BA]KU and ADClow[BA]SK were significantly different (p=0.004 and 0.006) with
thresholds of 8.25 and 2.25, respectively (higher parameter
values indicated shorter OS). Also, higher baseline volumes
(V) were indicative of shorter OS (p=0.009, threshold 772 ml).
Similar findings were seen at the follow-up time point, where
ADClow[DU]ME, ADClow[DU]KU, and ADClow[DU]SK where
significantly different between long-and short-term OS patients, with p-values of 0.004, 0.02, and 0.014, respectively.
Lower ADClow[DU]ME (threshold: 3.25 x10-3mm²/s), and higher
ADClow[DU]KU (threshold: 10) and ADClow[DU]SK (threshold:
2.3) were indicative of shorter OS. Again, higher lesion volumes
(V) during follow-up were indicative of shorter OS (p=0.009,
threshold 386 ml). As expected, the results for differentiating
between long- and short-term PFS were less encouraging,
with only ADCavg[BA]KU and ADClow[DU]ME nearing, but not
reaching, significant values (p=0.054 and 0.07, respectively).
Optimal thresholds of both parameters were 4.25 and 3.25
x10-3mm²/s, with lower ADCavg[BA]KU and ADClow[DU]ME
projecting shorter PFS.
Conclusion: Histogram analysis of ADC parameters during
early PCT of inoperable MPM patients can differentiate between
patients with long-term and short-term OS, although PFS
separation is less accurate. These findings show that first order
histogram analysis of DWI data could be a useful tool for personalized care in patients with inoperable MPM. However, these
preliminary data need confirmation in larger patient groups.
Keywords: Diffusion weighted resonance imaging, biomarker,
inoperable MPM, Histogram Analysis
MS03.05: CORRELATION OF CT SCAN BASED
TUMOR VOLUME MEASUREMENT TO ACTUAL
RESECTED TUMOR WEIGHT: A NEW T-FACTOR?
Isabelle Opitz1, Martina Friess1, Thi Dan Linh Nguyen-Kim2,
Thomas Frauenfelder2, Sven Hillinger1, Burkhardt Seifert3 , Ilhan
Inci1, Walter Weder1
Division of Thoracic Surgery, University Hospital Zurich, Zurich,
SWITZERLAND, 2Institute of Diagnostic And Interventional Radiology, University Hospital Zurich, Zurich, SWITZERLAND, 3Department of Biostatistics, Epidemiology, Biostatistics And Prevention
Unit, University of Zurich, Zurich, SWITZERLAND
1
Objectives: Tumor volume has been reported several times to
be a valuable prognosticator for malignant pleural mesothelioma (MPM) survival (Pass 1998, Opitz 2015). We wanted to
assess the precision of CT scan based preoperatively measured
tumor volume when correlated to the actual resected tumor
weight during macroscopic complete resection and their impact
on overall survival.
Methods: From October 2012 until November 2015 the tumor
weight of surgery specimens was measured in 27 patients
undergoing macroscopic complete resection. 26 patients were
male (96%), 25 MPM showed epithelioid type (96%) and the
median age at surgery was 66 years (range 41-77). Twenty-two
patients underwent induction chemotherapy prior to surgery.
In all 27 patients tumor volume was measured in the CT or
PET-CT scans performed before surgery as described previously (Frauenfelder 2011). Relations between tumor weight and
volume were analyzed using Pearson correlation. Tumor volume
and tumor weight were also tested for correlation with pT stage
using Spearman ranks correlation. Post-hoc comparisons
between stages were performed using the Mann-Whitney U
test. Association of dichotomized tumor volume and weight with
overall survival (OS) was evaluated using the log rank test.
Results: The median tumor volume assessed by CT scan
was 79 ml and the median tumor weight 520 g. The analysis
revealed a correlation between tumor volume and weight
(r=0.53, p=0.005). There was also a significant correlation of
tumor volume (p=0.001) as well as tumor weight (p<0.0005)
with the pT-stage (Figure 1). No significant association of tumor
volume and weight with OS was found but 82% of the cases
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ABSTRACT BOOK
were censored.
Figure 1: Box plot showing association of pT stage with tumor
volume (A) and weight (B) using Mann-Whitney U test.
Conclusion: The preoperative assessed tumor volume in CT
scan seems to be a valuable descriptor of actual tumor volume
to be resected and might be a reliable future T-descriptor.
Keywords: CT scan, tumor volume, tumor weight, TNM staging
MS03.06: DYNAMIC CONTRAST-ENHANCED CT
FOR THE ASSESSMENT OF TUMOR RESPONSE IN
MALIGNANT PLEURAL MESOTHELIOMA: A PILOT
STUDY
Eyjolfur Gudmundsson1, Sam Armato1, Zacariah E. Labby2,
Christopher M. Straus1, Feng Li1, Hedy Kindler1
Department of Radiology, University of Chicago, Chicago, IL,
UNITED STATES OF AMERICA, 2Department of Human Oncology, University of Wisconsin, Madison, WI, UNITED STATES OF
AMERICA
1
Objectives: Few investigations have been made into the use of
imaging-derived hemodynamic parameters for the assessment
of tumor response in malignant pleural mesothelioma (MPM)
patients. The objective of this study was to evaluate the utility of
dynamic contrast-enhanced computed tomography (DCE-CT) in
the assessment of MPM tumor response.
Methods: The standard CT imaging protocol for MPM was
modified to include a DCE-CT component, during which a 55mm axial extent of thoracic anatomy demonstrating notable
tumor burden was imaged at specific time points following
the start of contrast injection. The image-acquisition protocol
included two dynamic contrast-enhanced phases, one prior
to and one following a standard CT scan of the full chest. 16
patients were evaluated: eight on treatment, eight on observation. Each patient underwent two DCE-CT scans at approximately 3-month intervals. To capture tumor burden in each
scan, modified RECIST measurements were obtained manually
by a research radiologist, and CT-based volume measurements
were obtained using a semi-automated in-house method. To
define a region of interest for the computation of hemodynamic parameters, visible tumor was manually contoured on the
images from a single time point of the dynamic contrast-enhanced phases of each scan; these contours were automatically
propagated across all time points using a deformable image
registration technique. Perfusion, peak CT value enhancement,
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21
ABSTRACT BOOK
blood volume, and time to peak enhancement were calculated
from the contrast uptake curves obtained from each pixel within
the tumor contours. Mean changes in these parameters were
calculated between the two DCE-CT scans, and these parameter changes were compared between the on-treatment and
on-observation cohorts.
Results: Although changes in hemodynamic parameters were
not significantly different between the two patient cohorts
for any of the measured parameters, patients on treatment
demonstrated a mean relative decrease in blood volume and
perfusion (-14.2% and -17.2%, respectively) compared with
a mean relative increase in these parameters (+8.8% and
+27.0%, respectively) for patients on observation. No statistically significant correlation was found between relative change
in hemodynamic parameters and changes in tumor size, either
by modified RECIST or tumor volume. MS04: CELL AND VACCINE BASED THERAPY
MONDAY, MAY 2, 2016
14:15 – 15:45
MS04.02: EXPERIMENTAL MODELS OF HUMAN
MALIGNANT MESOTHELIOMA IN NOD SCID MICE
AND NUDE RATS FOR EVALUATION OF IN VIVO
VIROTHERAPY
Joëlle Nader1, Nicolas Boisgerault1, Carole Achard1, Tiphaine
Delaunay1, Myriam Robard2, Jean-François Fonteneau1,
Frédéric Tangy3 , Marc Grégoire1, Daniel L. Pouliquen1
UMR 892 INSERM / 6299 CNRS, Nantes, FRANCE, 2Plateforme MicroPICell, SFR F. Bonamy, Nantes, FRANCE, 3Unité de
Génomique Virale et Vaccination, Institut Pasteur, Paris, FRANCE
1
Geometric mean of relative changes Δ in DCE-CT
parameters from first scan to second scan for the
two patient cohorts. p-values were calculated using
a Student’s t-test.
DCE-CT Parameter
ΔTreatment
ΔObservation
p-value
Perfusion
-17.2%
+27.0%
0.14
Peak Enhancement
-8.4%
+1.1%
0.51
Blood Volume
-14.2%
+8.8%
0.21
Time to Peak
+0.2%
-17.2%
0.80
Conclusion: Hemodynamic parameters were computed from
DCE-CT scans acquired at two time points from MPM patients.
Observed differences in hemodynamic parameter changes
between patients on treatment and patients on observation
suggest that DCE-CT could be a useful imaging modality for
the assessment of tumor response. The significance of these
trends should be investigated through future studies with larger
numbers of patients and focused therapeutic regimens.
Keywords: imaging, CT, pleural mesothelioma, perfusion
Objectives: Oncolytic viruses are now considered as a new
therapeutic strategy against several cancers, as they are capable
of both preferential toxicity against tumor cells and simultaneous
activation of the host anti-tumor immunity. Previous studies have
demonstrated that attenuated strains of measles virus (MV) can
infect and kill different cell lines of human malignant mesothelioma (MM) in vitro. In this study, we present a human MM cell line
that produces tumors in the peritoneal cavity of immunodeficient
mice and rats, which represent two interesting models for the
evaluation of anti-tumor virotherapy in vivo.
Methods: The Meso 34 cell line belongs to a human biocollection of MM cell lines established from the pleural effusions of
patients and validated by the French MESR (n° DC-2011-1399,
CNIL n° 1657097). About 5x106 cells were injected intraperitoneally to NOD SCID mice and Nude rats. The MV Schwarz
strain and a modified strain with enhanced pro-apoptotic properties (MV-deltaC) were provided by Dr Frédéric Tangy (Institut
Pasteur). The viruses were injected i.p. with a single dose (2.105
TCID50) 8 weeks after tumor challenge. Animals were sacrificed
15 days after virus injection and tumor mass was evaluated.
Results: Eight weeks after tumor challenge, macroscopic tumors were present in both species as an omental cake together
with metastatic nodules attached to the diaphragm, liver and
spleen. A clear decrease of the tumor mass was observed in
MV and MV-deltaC groups. Tumor regressions were even more
important in the latter compared to the controls. Preliminary
histological analysis revealed a modification of tumor cell morphology and tumor density after virus infection, with numerous
zones of necrosis especially in the MV-deltaC group.
Conclusion: Our preliminary results demonstrate for the first
time that after one single intracavitary administration, both MV
and its genetically-modified variant infect and kill human MM
cells in vivo. Additionally, MV-deltaC, which shows enhanced
pro-apoptotic properties compared with MV, induced a more
significant decrease of the total tumor mass compared with
untreated mice. Meso 34 tumors transplanted in the nude rat
could also represent an interesting model to investigate the
effects of adoptive immunotherapy with human immune cells.
Keywords: Virotherapy, Animal model, Human cell line, Oncolytic measles virus
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ABSTRACT BOOK
MS04.03: TUMOUR SUPPRESSOR MICRORNAS
REGULATE PD-L1 EXPRESSION IN MALIGNANT
Marissa Williams1, Steven Kao1, Wendy A. Cooper2, Yuen Yee
Cheng1, Michaela B. Kirschner3 , Jason Madore2, Trina Lum4 ,
Anthony Linton1, Brian C. Mccaughan5, Sonja Klebe6 , Nico Van
Zandwijk1, Richard A. Scolyer7, Michael Boyer8 , Glen Reid1
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA, 2Sydney Medical School, The University of Sydney, Sydney,
NSW, AUSTRALIA, 3Division of Thoracic Surgery, University
Hospital Zurich, Zurich, SWITZERLAND, 4Department of Tissue
Pathology And Diagnostic Oncology, Royal Prince Alfred Hospital,
Sydney, NSW, AUSTRALIA, 5Sydney Cardiothoracic Surgeons,
RPA Medical Centre, Sydney, NSW, AUSTRALIA, 6Department of
Anatomical Pathology, Flinders Medical Centre, Adelaide, SA,
AUSTRALIA, 7Department of Medical Oncology, Concord Cancer
Centre, Sydney, NSW, AUSTRALIA, 8Chris O’brien Lifehouse,
University of Sydney, Camperdown, NSW, AUSTRALIA
1
Objectives: Programmed death 1 (PD-1) and its ligand PD-L1
have significant roles in suppressing host immune response in
many cancer types. As in other cancers,PD-L1 expression is
upregulated and associated with poor prognosis in malignant
pleural mesothelioma (MPM) but the mechanisms causing its
dysregulation are poorly understood. Characterization of the
mechanisms leading to PD-L1 upregulation could improve the
understanding of its dysregulation in MPM and give depth to its
prognostic significance.
Methods: Tissue Microarrays were constructed from formalin-fixed paraffin-embedded (FFPE) tissue blocks from patients
that underwent pleurectomy ± decortication (P/D). PD-L1 protein expression in patient and cell line samples was analysed
using a commercially available PD-L1 immunohistochemistry
assay. RT-qPCR was used to assess microRNA expression in tumour samples with specific Taqman probe-based assays. MPM
cell lines were reverse-transfected with synthetic microRNA
mimics and siRNAs using Lipofectamine RNAiMAX. RNA was
extracted using TRIzol and mRNA levels of PD-L1 and IRF-1
were determined using RT-qPCR and designed primers.
Results: We first established the prevalence of PD-L1 expression in a series of 72 MPM patients and found 18 (25%) had
positive PD-L1 staining that was more common in the non-epithelioid subtype (p=0.01), also, PD-L1 expression was associated with poor survival (median OS: 4 vs. 9.2 months, positive
and negative PD-L1 respectively; p<0.001) and its prognostic
significance was confirmed using the Cox Regression model after adjustment for gender, age and histological subtype (HR 2.2,
95% CI: 1.2-4.1; p<0.01). Reduced microRNA expression was
related to elevated PD-L1 levels in the MPM patient panel, with
previously identified tumour suppressor microRNAs in MPM
showing downregulation in PD-L1 positive tumours. The median
microRNA levels of miR-15b, miR-16, miR-193-3p, miR-195 and
miR-200c were shown to be significantly lower in PD-L1 positive
(N=13) than in the PD-L1 negative samples (N=50). miR-15a
and miR-16 are both predicted to target the 3’UTR region of
PD-L1, to characterize their regulatory affect, we restored their
expression in MPM cell lines which led to downregulation of
PD-L1 mRNA and protein expression. Transfection with miR193a-3p indirectly downregulated PD-L1 mRNA expression
via its regulation of IRF-1, a known transcriptional inducer of
PD-L1. This transcriptional regulation was further supported
by the reduction of PDL-1 mRNA upon transfection with IRF-1
specific siRNAs. Furthermore, transfection with miR-15a and
miR-16 were also shown to reduce IRF-1 expression in MPM cell
lines, suggesting both direct and indirect regulation of PD-L1
expression by tumour suppressor microRNAs in MPM.
Conclusion: This study has confirmed PD-L1 to be an adverse
prognostic indicator in MPM. Elevated PD-L1 expression in
MPM patient samples was correlated to downregulation of
tumour suppressor microRNAs that were shown to directly and
indirectly regulate PD-L1 expression in vitro.
Keywords: tumour suppressor, microRNA, mesothelioma,
PD-L1
MS04.04: PD-1+ T CELLS IN MESOTHELIOMA
EFFUSIONS INDUCE TUMOR PD-L1 EXPRESSION
MAKING THEM SUSCEPTIBLE TO AVELUMAB
MEDIATED ADCC
Swati Khanna1, Anish Thomas1, Daniel Abate-Daga2, Jingli
Zhang1, Betsy Morrow1, Seth Steinberg3 , Augusto Orlandi4 ,
Patrizia Ferroni5, Jeffrey Schlom6 , Fiorella Guadagni5, Raffit
Hassan1
Thoracic And Gastrointestinal Oncology Branch, National Cancer
Institute, Bethesda, MD, UNITED STATES OF AMERICA, 2Moffitt
Cancer Center, Tampa, UNITED STATES OF AMERICA, 3Biostatistics And Data Management Section, National Cancer Institute, Bethesda, MD, UNITED STATES OF AMERICA, 4Anatomic
Pathology, Dept Of Biomedicine And Prevention, University of
Rome, Rome, ITALY, 5Rome And Biodat (biomarker Discovery
And Advanced Technologies), Sr Research Center, University San
Raffaele, Rome, ITALY, 6Laboratory of Tumor Immunology And Biology, National Cancer Institute, Bethesda, MD, UNITED STATES
OF AMERICA
1
Objectives: To evaluate expression of programmed cell death 1
(PD-1) and PD ligand 1 (PD-L1) in patients with pleural and peritoneal mesothelioma, as well as their interactions in malignant
effusions and peripheral blood. In addition, we wanted to investigate the role of avelumab, a fully humanized IgG1 anti-PD-L1
antibody, in mediating antibody dependent cellular cytotoxicity
(ADCC) of PD-L1 expressing tumor cells by NK cells.
Methods: Formalin-fixed, paraffin-embedded (FFPE) tumor tissues from 44 peritoneal and 21 pleural mesothelioma patients
were tested for PD-L1 expression using anti-PD-L1 rabbit monoclonal antibody (MKP-1B-196-10-Merck-Serono). Staining was
recorded as positive if ≥5% of the tumor cells had membrane
PD-L1 expression. All autologous tumor cells and immune cells
(T, Monocytes and NK) were derived from either ascites or pleural fluid of mesothelioma patients. The PD-1 and PD-L1 positivity of T cells in paired blood and malignant effusion samples
(n=3) were evaluated by flow cytometry. For co-culture studies,
ascites from a mesothelioma patient (NCI-Meso29) was used
as a source of lymphocytes and tumor cells. Lymphoid cells
growing in suspension were tested for recognition of autologous tumor cells by IFN-γ release upon overnight co-culture.
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ABSTRACT BOOK
For ADCC experiments Carboxyfluorescein succinimidyl ester
stained NK cells were cultured with autologous or allogeneic
primary mesothelioma tumor cell lines (n=3; NCI-Meso21,
NCI-Meso29, NCI-Meso49) at different effector target ratio with
20 ng/mL of avelumab for 4 h. The cultured cells were stained
with 7-AAD and analyzed on a flow cytometer for detection of %
specific lysis of tumor cells.
Results: Out of 65 pleural and peritoneal mesothelioma
tumors examined, 41 (63%) were PD-L1 positive including 31
of 41 (76%) patients with peritoneal mesothelioma. A trend for
inferior overall survival in patients with PD-L1 positive tumors
was found, although the difference did not reach statistical
significance (median 23.0 vs. 33.3 months; p=0.35). The
fraction of cells expressing PD-L1 in malignant mesothelioma
effusions ranged from ~12 to 43%. Two of the 3 patients with
paired effusion and blood samples had high PD-1 expression
on both CD4+ and CD8+ T cells (23.8%, 42.1% vs. 13.1%,
6.43%, respectively) present in effusion compared to peripheral
blood. In addition, all 3 patients had higher PD-L1 expression
on CD3+ T cells present in malignant effusions compared with
those present in peripheral blood (7.5±2.6 % vs. 1.9±1.2%,
respectively). Autologous lymphocytes present in the malignant
effusion recognized tumor cells and induced IFN-γ-mediated
PD-L1 expression on the tumor cell surface. Out of 3 primary
mesothelioma cell lines tested, two (NCI-Meso21 and NCI-Meso29) were susceptible to ADCC by allogeneic and autologous
NK cells, respectively in presence of avelumab with specific
lysis of 12% and 23%, respectively at E:T ratio of 25:1.
Conclusion: PD-L1 is highly expressed on tumor tissue as well
as on malignant cells in ascites and pleural effusions of patients
with peritoneal and pleural mesothelioma. In addition, PD-1+ T
cells in malignant effusion induce tumor PD-L1 expression and,
in presence of the anti-PD-L1 antibody avelumab, are susceptible to NK cell mediated cytotoxicity.
Keywords: PD-1 PD-L1 checkpoint, Malignant effusions, Antibody dependent cellular cytotoxicity, mesothelioma
MS04.05: HARNESSING THE IMMUNE
SYSTEM-ADJUVANT IMMUNOTHERAPY FOR
MESOTHELIOMA
Methods: We developed a murine model of mesothelioma
debulking surgery to evaluate several clinically available immunotherapy approaches (immuno-gene therapy with Ad.mIFNa
and COX-2 inhibition with Celebrex) and to optimize dosing
and timing strategies. Outcomes were assessed for overall
survival and recurrent tumor burden. Flow Cytometry and in
vivo neutralization assays were utilized to determine immune
responses.
Results: As compared to mice receiving a control vector,
mice randomized to neoadjuvant Ad.mIFNa were associated
with a 64% reduction in tumor volume at post-operative-day
14 (420mm3 vs. 1183 mm3; p<0.01) and were associated with
an increase in median postoperative survival (34 days vs. 18
days, p =0.003). In vivo tumor neutralization assays demonstrated that the cytotoxic activity of CD8+ T-cells drove this
response. The addition of Celebrex to adjuvant Ad.mIFNa
improved potency and nearly tripled median post-operative
survival as compared to controls (48 days vs 17 days;p =0.002).
Additionally, 40% of mice receiving combination immunotherapy with surgery were cured vs. none in monotherapy
groups (p =0.03). Mice receiving both Ad.mIFNa and Celebrex
had markedly increased CD8+ T-cell trafficking in recurrent
tumors.
Conclusion: This evidence supports a new approach to
mesothelioma which includes multi-modal immunotherapy
with surgical debulking. We plan to use this preclinical data to
develop a Phase I Clinical Trial incorporating immunotherapy
with surgery for mesothelioma patients.
Keywords: Immunotherapy, mesothelioma, Surgery
MS04.06: INTRAPLEURAL MODIFIED VACCINE
STRAIN MEASLES VIRUS THERAPY FOR PATIENTS
WITH MALIGNANT PLEURAL MESOTHELIOMA - A
PHASE I TRIAL
Tobias Peikert1, Sumithra Mandrekar1, Aaron S. Mansfield1,
Virginia Van Keulen1, Steven Albelda2, Ileana Aderca1, Sephanie
Carlson1, Allen Dietz1, Mike Gustafson1, Robert Kratzke3 , Val
Lowe1, Fabien Maldonado4 , Julian Molina1, Manish Patel3 , Anja
Roden1, Angelina Tan1, Evanthia Galanis1
Mayo Clinic, Rochester, UNITED STATES OF AMERICA, 2University of Pennsylvania, Philadelphia, PA, UNITED STATES OF
AMERICA, 3University of Minnesota, Minneapolis, MN, UNITED
STATES OF AMERICA, 4Vanderbilt University, Nashville, TN, UNITED STATES OF AMERICA
1
Jarrod Predina1, Jane Keating2, Sarah Nims2, Ollin Venegas2,
Daniel Sterman3 , Sunil Singhal2, Steven Albelda1
University of Pennsylvania School of Medicine, Philadelphia,
UNITED STATES OF AMERICA, 2Thoracic Surgery, University of
Pennsylvania, Philadelphia, PA, UNITED STATES OF AMERICA, 34.
pulmonary, Critical Care And Sleep Medicine, NYU Langone
Medical Center, NY, NY, UNITED STATES OF AMERICA
1
Objectives: Our group has been interested in immunotherapy
for mesothelioma for nearly three decades. In our experiences,
the most important predictor of immunotherapy response in
human trials has been minimal disease at the time of drug delivery. With this in mind, we have hypothesized that “adjuvant”
immunotherapy with debulking surgery can improve immunotherapy efficacy and long-term response rates.
Objectives: Malignant pleural mesothelioma (MM) remains an
almost universally fatal disease with limited treatment options.
Preclinical models indicate the preferential oncolytic activity
of the modified vaccine strain measles virus (MV) carrying the
gene for the human sodium-iodine symporter (NIS) – MV-NIS.
Furthermore the intraperitoneal and intravenous administration of MV-NIS was recently found to be safe and potentially
effective in patients with refractory ovarian cancer and multiple
myeloma respectively. However, whether MV-NIS is directly
oncolytic or triggers an anti-tumor immune response in patients
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ABSTRACT BOOK
is unclear.
Methods: We conducted a Phase I dose escalation study with
3+3 design. MV-NIS was administered as first or second line
therapy via a tunneled intrapleural catheter to patients with
MM. MV-NIS dose ranged from 108 TICID 50 (Level 1) to 9 x
109 TICID 50 (Level 4). In the absence of dose limiting toxicity
and disease progression MV-NIS therapy was continued for up
to six cycles. MV-NIS infection and replication were monitored
by Iodine123 SPECT/CT as well as by RT-PCR and/or plaque-assay. Anti-tumor immunity was monitored in the blood and
pleural fluid and patients were followed clinically by chest CT
using the modified RECIST criteria for MM.
Results: Twelve patients (3 patients per dose levels 1-4)
received MV-NIS therapy. There were no dose limiting adverse
events and therapy was well tolerated. The best therapeutic
response was stable disease, which was achieved by 8/12
patients (67%). Overall survival demonstrated a median
survival of 449 days (~15 months) (5/12 patients are still alive).
Progression free survival was 62 days (~2 months). (Figure 1.)
MV infection and replication were detectable by RT-PCR and
plaque assay in the pleural fluid between 24-72 hours after
treatment. I123 SPECT-CT only demonstrated marginal viral gene
expression in a single patient treated with the highest dose
level. MV-NIS therapy effectively boosted pre-existing anti-MV
neutralizing antibody responses in the plasma and pleural fluid
of most patients. We observed a transient intense inflammatory
response in the pleural space after MV-NIS administration. In
addition activation of cellular and humeral immunity (induction
or boosting of anti-tumor antibody responses) was observed.
Conclusion: The intrapleural administration of MV-NIS is
safe, resulted in stable disease for 67% of patients and may be
associated with favorable overall survival in MM. While there is
only transient infection and viral replication, we have observed
the induction of anti-tumor immune responses. The study will
continue with a maximum tolerated dose expansion cohort.
Keywords: measles virus, virotherapy, tumor vaccine, mesothelioma therapy
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ABSTRACT BOOK
MS04.07: NIVOLUMAB IN MALIGNANT PLEURAL
MESOTHELIOMA (NIVOMES): AN INTERIM
ANALYSIS
Josine Quispel-Janssen, Marion Zimmerman, Wieneke Buikhuisen, Sjaak Burgers, Giulia Zago, Paul Baas
Thoracic Oncology, Netherlands Cancer Institute, Amsterdam,
NETHERLANDS
Objectives: Malignant pleural mesothelioma (MPM) is well
known for its resistance to therapy. No studies in second line
have yet resulted in an improvement of overall survival. Immunotherapy has attracted attention since lymphocyte infiltrates
are present in many cases of mesothelioma, and PD-L1 expression has been reported in 13% of epithelioid mesothelioma
and 38-100% of the sarcomatoid type. Nivolumab is a monoclonal antibody that binds PD-1 on activated immune cells and
disrupts binding of PD-1 to its ligand PD-L1, thereby preventing
down regulation of T-cells and augmenting the host antitumor
response. Here we describe the clinical results of an interim
analysis of a translational study (NivoMes trial).
Methods: In this single center, Simon two-stage, phase II
study, patients with progression after first line chemotherapy,
are treated with nivolumab 3mg/kg i.v. every two weeks. Primary endpoint is disease control rate (DCR) at 12 weeks. Secondary endpoints include overall survival and progression free
survival. Patients underwent repeat biopsies at start and after
3 courses of treatment for biomarker research. PD-L1 status is
not determined upfront. A total of 33 patients will be recruited.
An interim analysis is performed as planned.
Results: Eighteen patients were evaluated for the interim
evaluation after 12 weeks of treatment. Seven patients showed
disease control (39%): five patients had a partial response
(PR) and two stable disease. Two patients had pseudoprogression prior to a partial response. Nine patients had progressive
disease. Two patients showed a mixed response but continued
treatment because of clinical benefit. Currently, the follow
up is too short to determine whether the sites of growth are
due to real progression or pseudoprogression. Toxicity so far
was mostly mild and manageable. One patient presented with
progression after one course and discontinued treatment due
to toxicity (grade 3 nausea and grade 3 pneumonitis). He was
treated successfully with oral steroids and subsequently a PR
was observed. A second patient with initial progression was
hospitalized due to pneumonitis and pericardial tamponade. He
completely recovered after oral steroids and pericardial drainage. Upon retreatment, a partial response was noted which is
currently ongoing.
Conclusion: Preliminary results show promising activity of
nivolumab in malignant pleural mesothelioma and a manageable toxicity profile.
Keywords: Immune checkpoint inhibitor, nivolumab, anti-PD1
MS04.08: IMMUNE RESPONSES FOLLOWING
INTRAPLEURAL ADMINISTRATION OF ONCOLYTIC
HSV1716 IN PATIENTS WITH MALIGNANT
Joe Conner1, Kirsty Learmonth1, Lynne Braidwood1, Penella
Woll2, Chelsea Bolyard3 , Balveen Kaur3 , Kevin Blyth4
Virttu Biologics Ltd, Glasgow, UNITED KINGDOM, 2University of
Sheffield, Sheffield Teaching Hospitals, Sheffield, UNITED KINGDOM, 3Ohio State University, Columbus, OH, UNITED STATES
OF AMERICA, 4Respiratory Medicine, Queen Elizabeth University
Hospital, Glasgow, UNITED KINGDOM
1
Objectives: HSV1716 (Seprehvir) is an oncolytic immunotherapeutic herpes simplex virus type 1 mutant deleted in the gene
encoding the neurovirulence factor ICP34.5. Mutants lacking
ICP34.5 are selectively replication competent in cancer cells
and induce anti-tumour immune responses. Data supporting immune-mediated efficacy stimulated by treatment with
HSV1716 includes pre-clinical evidence of Th1 cytokine/chemokine responses that facilitate systemic anti-tumour immune responses via cytotoxic T cells that also reduce the establishment
of metastases and protect from re-challenge. Recent evidence
from mesothelioma patients post HSV1716 treatment supports
this immunotherapy activity with robust Th1 cytokine responses
detected in pleural fluids, evidence of immune cell infiltration
and activity and the development of a novel anti-tumour IgG
immune response.
Methods: A phase I/IIa trial to determine the safety and potential for efficacy of HSV1716 given intrapleurally to patients with
MPM is currently ongoing. Patients receive 1x107iu HSV1716
through their pleural catheter on one, two or four occasions a
week apart, in three separate patient cohorts. To date 10 patients have been treated, 3 in the one and two dose and 4 in the
four dose cohorts and HSV1716 has been well-tolerated with
few adverse events in any patients. Pleural fluid and plasma
samples have been collected pre- and post treatment and analysed to assess patient responses to HSV1716 administration.
Results: HSV1716 replicated/persisted in most patients with
HSV DNA detected in the pleural fluids for, in some cases, up
to 28 days post-administration. Increased levels of HMGB1 and
HSP70 were detected in pleural fluids during this time indicating the potential for immunological cell death associated with
HSV1716 oncolysis. Robust Th1 responses with increased IFNγ,
IP-10, MIG, I-TAC and TNFα were observed in most patients
after HSV1716 administration with additional IL-2, IL-10 and
IL-12 responses most prominent in patients receiving 4 doses.
There was evidence of immune cell infiltration into pleural fluids
after HSV1716 treatment and increased levels of Granzyme
B in pleural fluids indicate immune cell-mediated cytotoxicty.
Analysis of plasma samples indicated anti-HSV IgG responses
post-HSV1716 administration, particularly after 2 and 4 doses.
Analysis of pleural fluid samples also indicated anti-HSV IgG
responses post-HSV1716 administration. Crucially, in most
patients, there was a novel anti-tumour IgG response as
detected by immunoblotting against extracts from MPM cell
lines indicating additional, tumour-directed immune responses.
Further studies on the identities of the infiltrating immune cells
and their targets are ongoing.
Conclusion: Our trial demonstrates that oncolytic HSV1716
iMig2016.ORG
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ABSTRACT BOOK
has immunotherapeutic potential capable of inducing novel
anti-tumour immune responses in mesothelioma patients and
further studies are ongoing. This also confirms pre-clinical studies that clearly demonstrated HSV1716’s immunotherapeutic
mode of action.
Keywords: pleural fluids, Immunotherapy, Th1 responses,
anti-tumour immune responses
MS05:OPTIMUM DIAGNOSTIC PATHWAY FOR SUSPECTED MESOTHELIOMA
MONDAY, MAY 2, 2016
16:30 – 18:00
MS05.01: MESOBANK - TODAY’S BIOSPECIMENS
FOR TOMORROW’S MEDICINE
sample undergoes centralised frozen section/H&E to QC for
percentage tumour nuclei and necrosis. To date, 461 samples
(355 epithelioid, 68 biphasic; 38 sarcomatoid) from 83 patients
have undergone QC. In keeping with the heterogeneity of mesothelioma tumour/stroma, the percentage of tumour present in
each sample ranges from 6% to >75%. TMA construction (4-6
cores per case) has begun at Cancer Research UK Cambridge
Centre using surgical resection tissue blocks of 800 cases from
10 centres. 26 novel mesothelioma cell lines are available. The
original tumour cell type and patient gender/age are available
alongside cell immunoreactivity data, HLA typing and SEM for
most lines. All samples are barcoded at source for tracking and
storage. A secure web-based multi-user database has been
developed for sample and data collection.
Conclusion: High quality QC mesothelioma tissue, DNA, blood
derivatives, pleural fluid, TMA sections and cell lines, all of
which have linked-anonymised clinical data, are now being supplied to research groups on a cost-recovery basis. For enquiries
about tissue and data availability please contact www.mesobank.com. 1) Rintoul RC et al., Thorax 2015; Oct14
Keywords: Tissue banking, mesothelioma, Tissue microarray,
Cell lines
Robert C. Rintoul1, Doris M. Rassl2, Jacki Gittins1, Nick Maskell3 , John Edwards4 , Dean Fennell5, Peter Szlosarek6 , Richard
Booton7, Anoop Chauhan8 , Stefan J. Marciniak9
Thoracic Oncology, Papworth Hospital NHS Foundation Trust,
Cambridge, UNITED KINGDOM, 2Department of Pathology,
Papworth Hospital NHS Foundation Trust, Cambridge, UNITED
KINGDOM, 3Academic Respiratory Unit, University of Bristol,
Bristol, UNITED KINGDOM, 4Northern General Hospital, Sheffield, UNITED KINGDOM, 5Cancer Studies, University of Leicester,
Leicester, UNITED KINGDOM, 6Centre For Molecular Oncology,
Barts Cancer Institute & St. Bartholomew’s Hospital, London,
UNITED KINGDOM, 7University Hospitals South Manchester,
Manchester, UNITED KINGDOM, 8Portsmouth Hospital NHS
Trust, Portsmouth, UNITED KINGDOM, 9Cambridge Institute For
Medical Research (cimr), University of Cambridge, Cambridge,
UNITED KINGDOM
1
Objectives: MesobanK UK, funded by the British Lung Foundation and Mick Knighton Mesothelioma Research Fund was
set up in 2012 to provide quality assured mesothelioma tissue
to facilitate basic and translational research in mesothelioma.
Such work may lead to the development of personalised treatments for mesothelioma in the future1.
Methods: 1 Using standardized collection protocols and kits
we aim to collect 300 cases of fresh mesothelioma tissue,
whole blood, serum, plasma and pleural fluid linked to a clinical
data set with follow-up data from the UK National Cancer Registration Service. Germline DNA from whole blood is extracted by
the East Anglian Regional Genetics Laboratory. 2 To construct a
tissue microarray (TMA) using formalin fixed paraffin embedded
tissue with linked clinical data. 3 To develop at least 20 new
fully annotated mesothelioma cell lines MesobanK abides by all
relevant UK and EU legislation regarding the collection of tissue
and data. MesobanK is overseen by a Steering Committee and
a Research Advisory Board advises on applications for samples.
Results: 103 patients with confirmed mesothelioma (all
subtypes) have donated up to 5 tissue samples each (in RNA
Later™) with matched blood from 13 centres. Each tumour
MS05.02: HAPTOGLOBIN PHENOTYPE IS A
RISK FACTOR FOR MALIGNANT PLEURAL
MESOTHELIOMA
Kevin Lamote1, Sigurd Delanghe2, Ruben De Smet3 , Joris R.
Delanghe3 , Jan Van Meerbeeck4
Respiratory Medicine, Ghent University Hospital, Ghent, BELGIUM, 2Internal Medicine, Ghent University Hopital, Ghent, BELGIUM, 3Clinical Biology, Micriobiology And Immunology, Ghent
University Hospital, Ghent, BELGIUM, 4Department of Thoracic
Oncology, University Hospital Antwerp, Edegem, BELGIUM
1
asbestos-related rare tumor of the serous membranes of the
lungs. The pathogenesis of MPM is linked to asbestos-induced
chronic inflammation, oxidant formation, haemolysis and
subsequent haemoglobin release, potentiating oxidative injury.
Haptoglobin (Hp) serves as a major antioxidant by binding free
haemoglobin in order to prevent oxidative damage and lipid
peroxidation. The Hp-Hb-complex binds the CD163 receptor
on accumulated macrophages which induces the uptake of the
Hp-Hb-complex and the transfer of iron into the macrophages.
Dependent on the Hp-phenotype (Hp 1-1, Hp 2-1 and Hp 2-2),
this complexing and coupling to the CD163 receptor can be
divergent, leading to the additional formation of reactive oxygen
species (ROS) next to ROS directly induced by asbestos fibers
or released by inflammatory cells. We hypothesize that, dependent on the Hp phenotype, asbestos-exposed individuals can
have additional radical formation next to asbestos-induced radicals and could relate to the risk of MPM. In order to determine
the Hp-phenotype as a risk factor in MPM, this cross-sectional,
retrospective study compares the Hp-phenotype distribution in
MPM patients with controls from a European population.
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ABSTRACT BOOK
Methods: Hp-phenotyping was done on 118 archived serum
samples of MPM patients by starch gel electrophoresis. In
short, starch gel is prepared using 11.5% hydrolyzed starch
in a 0.1 M Tris citrate buffer. Electrophoresis is performed in
a 0.3 M borate buffer for 1 hour at 200V. Visualization of the
Hp-Hb-complexes is done by peroxidase staining. The frequencies of Hp types (Hp 1-1, Hp 2-1 and Hp 2-2) were compared
with those from 918 healthy control subjects by a Chi²-test
(Table I).
Results: Table I: Study results. Conclusion: Our results indicate an important role of the
Hp-phenotype in MPM pathogenesis suggesting that Hp 1-1
phenotypic persons are more prone for MPM development and
have a 74% increased risk of developing MPM. Apart from the
asbestos-induced radical formation, this finding underlines
the importance of oxidative stress in cancer development
and opens new perspectives for screening. Asbestos-exposed
individuals with the Hp 1-1 phenotype could then be subjected
to a more intensive monitoring and screening for early detection
and hence, lead to a better prognosis and management of the
disease.
Keywords: reactive oxygen species, oxidative stress, mesothelioma, haptoglobin
MS05.03: HMGB1 AND ITS ISOFORM ARE
SENSITIVE AND SPECIFIC BIOMARKERS TO
DETECT ASBESTOS EXPOSURE AND TO IDENTIFY
MESOTHELIOMA PATIENTS
Andrea Napolitano1, Daniel J. Antoine2, Laura Pellegrini1,
Francine Baumann1, Ian Pagano1, Sandra Pastorino1, Chandra
M. Goparaju3 , Kirill Prokrym3 , Claudia Canino3 , Harvey I. Pass3 ,
Michele Carbone1, Haining Yang1
University of Hawaii Cancer Center, Honolulu, HI, UNITED
STATES OF AMERICA, 2University of Liverpool, Liverpool, UNITED
KINGDOM, 3New York University, New York, NY, UNITED STATES
OF AMERICA
1
Objectives: To determine whether serum levels of High Mobility Group Box Protein-1 (HMGB1) could differentiate malignant
mesothelioma (MM) patients, asbestos-exposed individuals,
and unexposed controls.
Methods: Hyper-acetylated and non-acetylated HMGB1
(together referred to as total HMGB1) were blindly measured
in blood collected from MM patients (n=22), individuals with
verified chronic asbestos exposure (n=20), patients with benign
pleural effusions (n=13) or malignant pleural effusions not due
to MM (n=25), and healthy controls (n=20). Blood levels of
previously proposed MM biomarkers fibulin-3, mesothelin, and
osteopontin were also measured in non-healthy individuals.
Results: HMGB1 serum levels reliably distinguished MM
patients, asbestos-exposed individuals, and unexposed
controls. Total HMGB1 was significantly higher in MM patients and asbestos-exposed individuals compared to healthy
controls. Hyper-acetylated HMGB1 was significantly higher in
MM patients compared to asbestos-exposed individuals and
healthy controls, and did not vary with tumor stage. At the
cut-off value of 2.00 ng/ml, the sensitivity and specificity of
serum hyper-acetylated HMGB1 in differentiating MM patients
from asbestos-exposed individuals and healthy controls was
100%, outperforming other previously proposed biomarkers.
Combining HMGB1 and fibulin-3 provided increased sensitivity
and specificity in differentiating MM patients from patients with
cytologically benign or malignant non-MM pleural effusion.
Conclusion: Our results are significant and clinically relevant
as they provide the first biomarker of asbestos exposure and
indicate that hyper-acetylated HMGB1 is an accurate biomarker
to differentiate MM patients from individuals occupationally
exposed to asbestos and unexposed controls. A trial to independently validate these findings will start soon.
Keywords: HMGB1, mesothelioma, asbestos, biomarker
MS05.04: EXPRESSION OF MICRORNAS IN MPM
AS TOOL TO IDENTIFY NOVEL THERAPEUTIC
TARGETS AND DIAGNOSTIC/PROGNOSTIC
BIOMARKERS
Chiara De Santi1, Ombretta Melaiu2, Alessandra Bonotti3 , Luciano Cascione4 , Rudy Foddis5, Alfonso Cristaudo5, Marco Lucchi6 , Marco Mora7, Anna Truini7, Andrea Tironi8 , Bruno Murer9,
Renzo Boldorini10, Federica Gemignani1, Pierluigi Gasparini11,
Luciano Mutti12, Stefano Landi1
Department of Biology, University of Pisa, Pisa, ITALY, 2Immuno-Oncology Laboratory, Department of Paediatric Haematology/Oncology, Ospedale Pediatrico Bambino Gesù, Rome,
ITALY, 3cPreventive and Occupational Medicine, University
Hospital of Pisa, Pisa, ITALY, 4dLymphoma and Genomics
Research Program, Institute of Oncology Research, Bellinzona,
SWITZERLAND, 5Department of Translational Research and of
new Technologies in Medicine and Surgery, University of Pisa,
Pisa, ITALY, 6Division of Thoracic Surgery, Cardiac and Thoracic
Department, University of Pisa, Pisa, ITALY, 7IRCCS H. San Martino-IST Genova, Genova, ITALY, 8hSection of Anatomic Pathology,
Oncology and Experimental Immunology, Department of Molecular and Translational Medicine, University of Brescia, Brescia,
ITALY, 9Azienda ULSS 12 Veneziana, Venezia, ITALY,10Department
1
iMig2016.ORG
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ABSTRACT BOOK
of Health Sciences, School of Medicine, University Hospital
Maggiore della Carità, Novara, ITALY, 11Department of Molecular Virology, Immunology and Medical Genetics, Ohio State
University Wexner Medical Center and Comprehensive Cancer
Center, Columbus, OH, UNITED STATES OF AMERICA, 12School of
Environment and Life Sciences, University of Salford, Manchester,
UNITED KINGDOM
Objectives: Malignant pleural mesothelioma (MPM) is one
of the most aggressive human cancers and miRNAs can play
a key-role for this disease. In order to broaden the knowledge
in this field, the miRNA expression was investigated in a large
series of MPM to discover new pathways helpful in diagnosis,
prognosis and therapy.
Methods: We employed nanoString nCounter system for miRNA profiling on 105 MPM samples and 10 healthy pleura. The
analysis was followed by the validation of the most significantly
deregulated miRNAs by RT-qPCR in an independent sample
set. Kaplan-Meier curves were used to explore the association between miRNA expression and overall survival (OS). In
silico analyses were also performed to understand the pathways
involving the evaluated miRNAs.
Results: We identified 63 miRNAs deregulated in a statistically
significant way. The top five significant were: miR-185-5p, miR197-3p, miR-299-5p, miR-337-3p, and miR-485-3p. MiR-185,
miR-197, and miR-299 were confirmed differentially expressed,
after validation study. In addition, the results of the microarray
analysis corroborated previous findings concerning miR-15b-5p,
miR-126-3p, and miR-145-5p. DIANA-microT-CDS highlighted
5 putative targets in common between two miRNAs. We also
identified a 2-miRNA signature (Let-7c-5p and miR-151a-5p)
related to overall survival.
Conclusion: With the present work we showed that the pattern
of miRNAs expression is highly deregulated in MPM. We also
suggested that alterations in miRNA expression could modify
cell pathways regulation, allowing the discovery of new druggable targets. Moreover, a two-miRNA signature can be a new
useful tool for prognosis in MPM.
Keywords: mirnas, Survival, microarray, target prediction
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ABSTRACT BOOK
MS05.05: THE ROLE OF MULTIDISCIPLINARY
DIAGNOSTIC/THERAPEUTIC PATHWAY IN THE
CASE-MANAGEMENT OF SUSPECTED PLEURAL
MESOTHELIOMA
Filippo Lococo1, Cristian Rapicetta1, Roberto Piro2, Carla Galeone1, Patrizia Ciammella3 , Sofia Taddei2, Debora Formisano4 ,
Lorenzo Agostini2, Francesca Zanelli5, Francesco Falco2, Sara
Tenconi6 , Angelina Filice7, Giorgio Sgarbi1, Massimiliano Paci1,
Luigi Zucchi2
Chirurgia Toracica, Arcispedale Santa Maria Nuova-IRCCS, Reggio Emilia, ITALY, 2Pneumologia, Arcispedale Santa Maria Nuova-IRCCS, Reggio Emilia, ITALY, 3Radiation Oncology, Arcispedale
Santa Maria Nuova-IRCCS, Reggio Emilia, ITALY, 4Scientific
Directorate, Arcispedale Santa Maria Nuova-IRCCS, Reggio
Emilia, ITALY, 5Oncology, Arcispedale Santa Maria Nuova-IRCCS,
Reggio Emilia, ITALY, 6University Hospitals of Leicester, Leicester,
UNITED KINGDOM, 7Nuclear Medicine, Arcispedale Santa Maria
Nuova-IRCCS, Reggio Emilia, ITALY
1
Objectives: There is significant variation between physicians in
care patterns of patients affected by malignant pleural mesothelioma (MPM). In the emerging era of “systems/network
medicine”, the degree of complexity of such disease requires
an integrative and multidimensional approach. The aim of this
study is to compare the management of MPM-patients, before
and after the introduction of a structured multidisciplinary
diagnostic/therapeutic pathway (MDTP).
Methods: We introduced MDTP in May 2012.; a dedicate “case
manager” (usually pulmonologist) accounted for the entire
pathway of the patient with suspected MPM (Figure-1). If diagnosis of MPM was confirmed, a specific multidisciplinary team
of experts discussed on clinical case, assessed the staging,
established and directly planned the best therapeutic proposal
according with the current ESMO-Clinical-Practice-Guidelines.
All clinical, surgical, pathological and follow-up data were
recorded into a dedicated prospective database (from 01/2010
to 06/2014), enrolling 75 consecutive MPM-patients: 48 before
the introduction of MDTP (Group pre-MDTP) and 27 thereafter
(Group post-MDTP). A comparative analysis was performed
evaluating two main indicators : 1) “Diagnosis Time” (D-Time):
the interval between the “first access” of the patient and
diagnosis; 2) “Treatment Time” (T-Time): the interval between
diagnosis and treatment.
Results: The main characteristics of the patients did not differ
between the two groups (see Table 1). In pre-MDTP, the median
“D-Time” and “T-Time” were 22 and 26.5 days, respectively.
After the introduction of the MDTP we observed a reduction of
both indicators (median “D-Time” and “T-Time” of 17 and 21
days, respectively), despite such differences did not reach the
statistical threshold.
Conclusion: Despite only preliminary, our data suggest that
a structured multidisciplinary diagnostic/therapeutic pathway
may help the management and clinical assistance of MPM-patients, shortening the diagnostic and therapeutic delay. Figure 1 Flowchart of the MDTP in suspected MPM-patients iMig2016.ORG
30
ABSTRACT BOOK
Table 1: The main characteristics of the population and compar-
ative analysis between pre-MDTP and post-MDTP mesothelioma nursing support and access to treatment and
clinical trials.
Methods: The regional thoracic oncology centre in Manchester, The University Hospital of South Manchester has launched
a Regional Mesothelioma MDT to serve the needs of the North
West Populatuion. The core MDT members are representatives
from Respiratory Medicine, Thoracic Surgery, Medical Oncology, Thoracic Radiology, Thoracic Histopathology, Cancer Nurse
Specilaists, Research Support and Administration; all with a
specialist interest in mesothelioma.
Results: The patinet pathway for our Regional Mesothelioma
MDT is outlined below. Keywords: Malignant pleural mesothelioma, structured multidisciplinary diagnostic/therapeutic pathway, multidisciplinary
team, case manager
MS05.06: LAUNCHING THE NORTH WEST
REGIONAL MESOTHELIOMA MDT; DEFINING THE
PATIENT PATHWAY
Conclusion: The North West Mesothelioma MDT has been
launched in Manchester following the pathway above. Outcome
data audited against the standards set out in the National Mesothelioma Audit will be presented in poster form at IMIG 2016.
Keywords: mesothelioma, MDT
Lorraine Creech, Anshuman Chaturvedi, Marie Kirwan, Sue
Jackson, Rajesh Shah, Paul Taylor, Matthew Evison, Jayne
Holme
Manchester Thoracic Oncology Centre, University Hospital of
South Manchester, LT, UNITED KINGDOM
Objectives: The North West of the United Kingdom has a high
prevalence of mesothelioma. There is a clear need to provide
multi-discplinary expertise for the region to facilitate accurate
staging, histological confirmation, tumour sub-typing, specialist
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ABSTRACT BOOK
MS06:ASBESTOS CONTROL
MONDAY, MAY 2, 2016
16:30 – 18:00
MS06.01: TURKEY NATIONAL MESOTHELIOMA
SURVEILLANCE PROGRAM AND ASBESTOS
CONTROL
Muzaffer Metintas
Department of Chest Diseases, ESKISEHIR OSMANGAZI UNIVERSITY MEDICAL FACULTY, ESKISEHIR, TURKEY
Objectives: Malignant mesothelioma is an important health
problem because of environmental and occupational asbestos
and erionite exposure in Turkey. However, there is no sufficient
data about surveillance of mesothelioma in Turkey. Between
2012 and 2015, Turkey National Mesothelioma Surveillance
Program and Turkey Asbestos Control Strategic Plan (TAKSAP)
were prepared with participation of hospitals from 30 provinces, where mesothelioma is endemic, and support of Turkey
Public Health Agency. The aim of the program was to determine
incidence of mesothelioma for Turkey, determine risk of cities
regarding environmental asbestos exposure and develop strategies to solution the problem.
Methods: In hospitals from 30 provinces, the patients diagnosed with “mesothelioma” under the code of C45.0-C45.9
between 2008 and 2012 were identified with approbation. The
cases were checked in the Central Register System too. In this
study, “from case to the field method” has been used. After
obtaining the final records of the cases with mesothelioma, the
cases born in villages/rural areas were determined; the villages
where these cases were born were identified as “villages required to be examined in terms of asbestos exposure risk”. The
soil samples from these villages were examined in the TUBITAK
Marmara Research Centre Material Institute for mineral analysis with x-rD (x-ray diffactometer). Direct Standardized Average
Annual Mesothelioma Incidence Rates (AMIRs) were calculated
from WHO standard population, 2002. Estimates the population exposed to standardized incidence ratios were calculated
by multiplying the standard population rates with the ratio of
expected values to the observed values, and standard error was
calculated as the inverse fraction of the square root of observed
value (1 / √ observed value).
Results: The numbers of mesothelioma cases confirmed were
5,617 according to TAKSAP within 5 years. The crude incidence rates of mesothelioma were 1.53/100,000 for all cases,
1.75/100,000 for men and 1.30/100,000 for women. AMIRs
were 3.79/100,000 for all cases, 2.28/100,000 for men, and
2.94/100,000 for women. The crude incidence rates of mesothelioma were 73.42/100,000 for all cases, 79.94/100,000 for
men and 66.92/100,000 for women in villages where asbestos
exposure was continuing. The crude incidence rates of mesothelioma were 26.94/100,000 for all cases, 32.98/100,000 for
men and, 20.87/100,000 for women in villages where asbestos
exposure was stopped. The 98.3% of mesothelioma cases were
from 30 provinces where TAPSAP was organized. The first four
provinces were Elazığ (RR 34.74), Eskişehir (25.58), Diyarbakır
(14.08), Tokat (8.01), when provinces were sorted in terms of
mesothelioma risk. The first four provinces were Elazığ (RR
18.21), Eskişehir (9.07), Diyarbakır (8.37) and, Tunceli (7.60),
when RR were sorted according to birth villages of cases.
158,068 people live in the 397 villages where asbestos exposure is continuing. 286,510 people live in the 174 villages where
there was asbestos exposure in the past. It is expected that
17,961 new cases of mesothelioma will emerge among exposed
population between 2013 and 2033.
Conclusion: We determined that mesothelioma due to environmental and occupational asbestos exposure in Turkey is a more
serious problem than previously anticipated. Turkish Mesothelioma Working Group Coordinators: Muzaffer Metintas1 and
Hasan F Batırel2. 1Eskisehir Osmangazi University Lung and
Pleural Cancers Application and Research Center, Eskisehir-Turkey, [email protected]. 2Marmara University Medical Faculty Department of Thoracic Surgery, İstanbul-Turkey,
[email protected]) Researchers: Drs. Abdurrahman Abakay,
Sedat Altın, Güntülü Ak, Şule Akçay, Hasan Bayram, Mehmet
Bayram, Serdar Berk, Mehmet Bilgin, Nilgün Yılmaz Demirci,
Figen Deveci, İsa Döngel, Ahmet Erbaycu, Dilek Ernam, Sebahat Genç, Murat Gültekin, Ezgi Hacikamiloğlu, Hüseyin İlter,
Selahattin Kadir, Hasan Kahraman, Mehmet Karadağ, Özkan
Kaan Karadağ, Talat Kılıç, Gamze Kirkıl, Berna Kömürcüoğlu,
Selma Metintaş, Arzu Mirici, Ömer Özbudak, Sibel Özkurt,Önder
Öztürk, Dursun Tatar, Engin Tutkun, Umran Toru, Toros Selçuk,
Zehra Seyfikli, Nazan Şen, Abdurrahman Şenyiğit, Gaye Ulubay,
Huseyin Yalcin, Ülkü Yılmaz, Adil Zamani
Keywords: incidence, mesothelioma, asbestos, environmental
exposure
MS06.02: MESOTHELIOMA MORTALITY AT 10YEAR FOLLOW-UP IN THE ATOM 002 SCREENING
STUDY
Ornella Belvedere
Dept. Of Oncology, York Hospital, York, UNITED KINGDOM
Objectives: We previously reported the results of a prospective, non-randomized trial of low-dose computed tomography
(LDCT) screening in 1,045 asbestos exposed subjects already
under surveillance at the local Occupational Health Unit with
annual clinic review and chest X-ray (ATOM002 study); overall,
nine early stage lung cancers but no malignant mesotheliomas
were detected in the prevalence screening performed between
February 2002 and October 2003 (Fasola et al., The Oncologist 2007; 12:1215). Here, in the context of the ATOM002 study,
we explore the impact of the participation in a screening study
on overall, all cancer, lung cancer and mesothelioma specific
mortality.
Methods: After an incidence screening LDCT at one year, the
ATOM002 subjects were returned to the Occupational Health
surveillance program. At 10-year follow up, cause of death data
were obtained from the regional population registry, classified
according to the ninth revision of the International Classification
of Disease (ICD-9). Standardized mortality ratios (SMR) were
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ABSTRACT BOOK
calculated for the ATOM002 participants and nonparticipants
using the regional and Italian mortality rates. Univariate and
multivariate analyses were performed to assess the relationship
between mortality and smoking status, age at start of follow-up,
level of exposure to asbestos, Charlson-Quan comorbidity index
and participation in the ATOM 002 study; all these variables
were included in the Cox model.
Results: Within the cohort of 2,433 asbestos-exposed subjects
enrolled in the public health surveillance program at the Occupational Health Unit in Monfalcone, Italy in 2002, we compared
the 10 year overall, cancer-related, lung cancer and pleural mesothelioma specific mortality of the participants and nonparticipants in the ATOM002 study (n = 926 and 1,507, respectively).
There was no difference in overall cancer mortality (HR=0.97,
95% CI 0.62-1.50) or pleural mesothelioma mortality (HR=0.86,
95% CI 0.31-2.41) between the ATOM002 participants and
nonparticipants. However, mortality from all causes (HR=0.61,
95% CI 0.44-0.84) and lung cancer mortality (HR=0.41, 95%
CI 0.17-0.96) were significantly reduced among subjects who
participated in the ATOM002 study.
Conclusion: The role of mesothelioma screening in asbestos
exposed subjects remains unclear, with most authorities currently advising against screening, mainly based on the lack of
effective intervention even for early disease. Our findings do not
support LDCT based screening for mesothelioma in asbestos
exposed subjects but warrant further investigation of LDCT
screening for lung cancer in this high-risk population.
MS06.03: FACTORS INFLUENCING THE ASBESTOS
BODIES AMONG PLEURAL MESOTHELIOMAS
AND LUNG CANCERS EXAMINED FOR RETAINED
ASBESTOS FIBRES
Paolo Girardi1, Anna B. Somigliana2, Pietro G. Barbieri3 , Enzo
Merler1
analysed for AF by means of a Scanning Electron Microscopy
equipped with X-rays fluorescence microanalyzer at 12,000
magnification for fibres longer than 1 μm, after ashing at low
temperature, and for AB by means of an Optical Microscope at
500 magnification, after chemical digestion with hypoclorite.
Whereas the MPM samples were derived from pleuro-pneumonectomies or autopsies, LC samples were because of autopsies
ordered by a coroner in subjects known to have been shipyard
workers. All subjects have been investigated and classified for
probability and periods of exposure to asbestos. For 121 subjects each fibre counted at SEM was defined for its length and
diameter and a fibre volume (mass) was calculated. Uni- and
multi-variate linear regression analyses were used to evaluate
the relationship between AB and AF and influencing factors.
Results: Out of 154 subjects (145 males) under study, 144 have
been classified with a certain occupational asbestos exposure
(97/104 for MPM; 47/50 for LC) with a long time since ceased
exposure (mean 28.3 years). Overall, the Geometric Mean (GM)
was 2,7 (95% CI 2.0-3.6) million AF/g dry tissue (dt) and 29,500
AB/g dt (95% CI 20,900-41,800). A strong association was
found between the log10 AB and log10 AF among occupational
exposed (Pearson Correlation Index, PCI=0.78), less among
non occupational (PCI=0.53). The multivariate regression
analysis showed that dimension and fibre type influenced the
AB lung burden: a clear positive association was observed with
AB count and both fibre mass and prevalent presence (>80%)
of amphiboles among AF. Time since last exposure positively
influenced the AB count: every 1-year increment since the end
of the exposure implies an increase of 2.4% (95% CI 0.8-4.0) of
the AB count.
Conclusion: The AB burden increases with the amount of
retained AF composed, among the subjects under study, of high
percentages of amphiboles. The AF mass correlates with AB
count, suggesting a role of the fibre dimension in the endless
sequences of the inflammatory process induced by fibres. Not
only amphiboles but also chrysotile fibres, even if with definitively a minor weight, are involved in the development of AB.
The time factors are crucial for the formation of AB.
Keywords: asbestos fibres, Lung cancer, mesothelioma,
asbestos bodies
Local Health Authority Of Padua, Venetian Mesothelioma Registry, Occupational Health Unit, Padova, ITALY, 2Centre of Electronic
Microscopy, Lombardy Environmental Protection Agency, Milan,
ITALY, 3Retired; formerly, Mesothelioma Registry, Occupational
Health Unit, Local Health Authority of Brescia, Brescia, ITALY
1
Objectives: A marker of exposure to asbestos is the detection
of asbestos fibres (AF) and asbestos bodies (AB) in the lungs
of subjects. Factors such as the amount, type, dimension of
the AF reaching the distant part of the lungs contribute to
the development of AB. AB develop, after undefined lengths
of time, in the framework of an inflammatory process as the
result of a biotransformation of longer asbestos fibres, usually
amphiboles. Because AB can be observed at the resolution of
an Optical Microscope, they have been more often investigated,
usually after a chemical digestion of a tissue sample, but with
a poor standardization of methods. We evaluate the correlation
between retained AF and amount of AB, by asbestos exposure
and time-related variables.
Methods: Freeze dried lung samples from 104 Malignant Pleural Mesothelioma (MPM) and 50 Lung Cancer (LG) have been
MS06.04: AN ECOLOGICAL ANALYSIS OF
COHORTS WITH ENVIRONMENTAL AND
OCCUPATIONAL MINERAL FIBER EXPOSURE
Selma Metintas1, Guntulu Ak2, Muzaffer Metintas2
Lung And Pleural Cancers Research And Clinical Center And
Medical Faculty Department of Public Health, Eskisehir Osmangazi University, Eskisehir, TURKEY, 2Lung And Pleural Cancers
Research And Clinical Center And Medical Faculty Department
of Chest Diseases, Eskisehir Osmangazi University, Eskisehir,
TURKEY
1
Objectives: Exposure to asbestos and erionite are well established etiological factors for the development of malignant
mesothelioma (MM). Exposure to asbestos is classified in three
iMig2016.ORG
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ABSTRACT BOOK
groups, i.e. occupational, environmental, and para-occupational, with differential etiologic roles. The aim of the present study
was to assess the factors associated with MM incidence using
a logistic regression model based on the reported incidence of
MM in these exposure groups
Methods: A total of 21 cohort datasets were retrieved from a
total of 39 publications regarding the MM incidence with asbestos or erionite exposure. Of these cohorts 10, 17, and 3 represented environmental, occupational, and para-occupational
populations, respectively. Data were analyzed using a multiple
linear regression analysis model with SPSS 15.0 statistical
software pack. The natural logarithm of MM incidence as the
dependent variable for MM incidence was determined. Independent variables included the gender, median exposure dose (f/
mL) for the fiber in question, cumulative exposure dose (f/mLyears), latent period, median age, age at the time of initial exposure, the type of the fiber, type of exposure, or the nature of the
exposure (continuous vs. interrupted). Dummy variables were
defined for the fiber type and exposure type. Regression model
was extended using the subject-year data for the cohorts.
Results: In studies examined, the reported incidence for MM
ranged between 7,27 and 1267,00 per 100,000 population, with
a median incidence of 82.7. In the environmental cohorts, the
corresponding figures for males and females were 298,10 (minmax: 11,20 – 639,00) and 400,90 (min-max: 9.40-1267,00), in
the occupational cohorts, the corresponding figures for males
and females were 43,69 (7,80-206,25) and 82,70 (15,40245,89). There were two para-occupational cohorts, for these
cohorts the corresponding figures were males and females
7,27 to 91,00 and 46,26 to 57,0, respectively. The median
exposure doses to asbestos in the environmental and occupational groups were 47,0 f/mL (20,8-60,30) and 19,0 (0,65-36,0),
respectively. Modelling could not be performed due to small
sample size in the para-occupational groups. The median
cumulative exposure doses in the environmental and occupational exposure cohorts were 5.40 f/mL-years (2,60-8,70)
and 20,78 f/mL-years (3,60-36,25), respectively. The median
duration of the latent period in environmental and occupational
asbestos exposure groups were 55,30 (48,0-60,70) and 36,10
(24,60-47,60) years, while the median of the median ages were
53,0 years (48,0 – 60,70) and 62,90 years (50,20 – 69,80),
respectively. The median age at the time of first exposure was
0 in the environmental cohorts and 25,60 years (18,0 – 30,60)
in the occupational cohorts. In the multiple regression analysis, the following were associated with MM incidence in the
overall cohort: the median duration of exposure β (95%CI)
-0.80 (-0.105; -0.055) (p<0.001), cumulative exposure dose
0.028 (0.013-0.043) (p<0.001), exposure to erionite type fiber
1.916 (0.040-3.791) (p=0.045), continuous exposure 2.93
(0.287-5.58) (p=0.030). In the model R2 was 0.771 and F was
29,371 (p<0.001). Factors associated with MM incidence in the
environmental exposure group included the following: gender β
(95%CI) -2.082 (-2.29 to -1.87) (p<0.001), median duration of
exposure -0.30 (-0.31 to -0.28 ) (p<0.001), cumulative exposure
dose 0.36 (0.29-0.43) (p<0.001), exposure to erionite type
fiber 3.05 (3.41-2.68) (p<0.001), continuous exposure 8.88
(8.56-9.20) (p<0.001), and the median age of the patient -0.38
(-0.430 to -0.33 ) (p<0.001). R2 was 0.998 and F was 7668,178
for the model (p<0.001). The following emerged as the factors
associated with MM incidence in occupational exposure groups:
median duration of exposure β (95%CI) -0.07 (-0.095 to -0.036)
(p<0.001), cumulative exposure dose 0.031 (0.012-0.049)
(p=0.001), and single exposure fiber type of asbestos -1.12
(-1.69 to -0.533) (p<0.001). R2 was 0.686 and F was 20,545 for
the model (p<0.001).
Conclusion: Despite well-established etiological factors for
MM, the need remains for better defining the epidemiological
associations to the characteristics of asbestos exposure, since
a number of factors such as the type and type of asbestos exposure, age at the onset of exposure, and duration of exposure
appear to be related with the incidence.
Keywords: incidence, epidemiology, Erionite, asbestos
MS06.05: THE IMPACT OF GEOGRAPHIC AND
SOCIOECONOMIC FACTORS ON PROGNOSIS AND
TREATMENT PROVISION IN MALIGNANT PLEURAL
MESOTHELIOMA
Anthony Linton1, Matthew J. Soeberg1, Richard Broome2, Nico
Van Zandwijk1
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA, 2Public Health Observatory, Sydney, NSW, AUSTRALIA
1
Objectives: The impact of clinico-pathological factors including age, gender and histological subtype on the prognosis on
malignant pleural mesothelioma (MPM) are well understood.
However the effect of socio-economic and geographic factors
are less certain. Whilst the majority of Australian’s reside in
major cities, a significant proportion of patients are located in
regional and remote areas, where access to clinical services
may be limited. We analysed the relation between geographic
and socio-economic factors upon survival and treatment provision in a large series of patients from New South Wales.
Methods: All patients registered with the NSW Dust Diseases Board (2002-2009) diagnosed with MPM were assessed.
Geographic remoteness, distance from oncological multidisciplinary teams (MDT) and index of relative socio-economic
advantage and disadvantage(IRSAD) , were assessed with
known prognostic factors using Kaplan Meir and Cox-regression
analysis. Chi-square testing compared categorical variables
to analyse impact of these factors upon clinical features and
treatment received.
Results: We identified 910 patients: 90% male, histology (epithelioid 60%;non-epithelioid 30%),geographic remoteness (major city 67%; regional or remote 33%), distance to MDT (<10km
65%, <50km 92%), IRSAD (above average 50%, below average
50%). Median overall survival was 10.0 months. On multivariate
analysis age >70 (HR=1.39), male gender (HR=1.37), non-epithelioid histological subtype (HR.2.19) and IRSAD status by
decreasing quintile (HR=1.07) were independent prognostic
factors. Trend to improved survival when residing in major cities
(10.6 vs 8.8 months;p=0.162) and within 50km of MDT (10.3 vs
7.8 months;p=0.539). Patients geographic location and distance
to MDT did not impact chemotherapy, adjuvant radiotherapy
or extrapleural pneumonectomy provision. Socioeconomically
disadvantaged patients were significantly less likely to receive
chemotherapy (40.3% vs 47.7%; p=0.032)
iMig2016.ORG
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ABSTRACT BOOK
Conclusion: Socioeconomic disadvantage was an independent
and significant prognostic factor for MPM in NSW Australia
despite access to ‘universal’ health care. A significant reduction
in chemotherapy utilisation was also noted. A trend to improved
survival was noted in patients residing in major cities within
closer proximity to oncology units however this was not statistically significant. Reassuringly, treatment provision did not differ
in patients regardless of geographic location. Socioeconomic
factors appear to be a greater cause of disparity in mesothelioma outcomes in comparison to geographic factors. Further
prospective research analyzing specific factors including
comorbidity, income, and individual preference will be required
to better understand these findings.
Keywords: prognosis
tional association of pleural MM with unknown primary cancer
restricted to follow-up ≥1 year, suggesting shared genetic or
non-genetic risk factors.
Keywords: pleural mesothelioma, Peritoneal mesothelioma,
second primary cancers, Malignant mesothelioma
MS06.07: MESOTHELIOMA INCIDENCE IN
SWEDEN - WHY DOES IT NOT GO DOWN?
Gunnar Hillerdal
Pulmonary, Gavle Hospital, GAVLE, SWEDEN
MS06.06: RISK OF SECOND PRIMARY CANCERS
AFTER MALIGNANT MESOTHELIOMA AND VICE
VERSA
Tianhui Chen1, Elham Kharazmi2, Jianlin Lou1, Xing Zhang1,
Kristina Sundquist3 , Kari Hemminki2
Institute of Occupational Diseases Prevention, Zhejiang Academy
of Medical Sciences, Hangzhou, CHINA, 2Division of Molecular
Genetic Epidemiology, German Cancer Research Center (DKFZ),
Heidelberg, GERMANY, 3Lund University, Malmö, SWEDEN
1
Objectives: Investigations on risk of second primary cancers
(SPCs) after malignant mesothelioma (MM) and vice versa have
not been reported. We aimed at investigating risk of specific
SPCs after MM and vice versa.
Methods: Patients diagnosed with pleural MM and peritoneal
MM in Sweden during 1997-2012 were selected, respectively.
Standardized incidence ratios (SIRs) were used to assess risk of
specific SPC after MM, compared to risk of the same first cancer in the Swedish general population. Calculations were also
performed in a reverse order by assessing risk of second MM
after any first cancers and same methodology was adopted.
Results: Among survivors of 3,672 pleural MM and 895 peritoneal MM, overall 113 and 28 SPCs were recorded, respectively,
while reverse analyses included overall 431 second pleural and
88 peritoneal MMs after any first cancers. Elevated SIR after
pleural MM was observed for total combined SPCs (SIR=1.4;
95%CI:1.1-1.6), ovarian [4.7 (1.3-12)] and kidney cancers [4.4
(2.0-8.3)], while reverse analyses found elevated SIR for second
pleural MM after total combined [1.2 (1.1-1.3)], connective
tissue [4.5 (1.7-9.9)], kidney [2.3 (1.3-3.9)] and lung cancers [1.9
(1.2-2.9)]. The bidirectional association of pleural MM with kidney cancer was restricted to follow-up <1 year [5.4 (2.0-12) and
4.9 (2.0-10), respectively] and with total combined cancers was
restricted to follow-up ≥1 year [1.4 (1.1-1.8) and 1.2 (1.1-1.3),
respectively; considering unknown primary cancer: 3.9 (1.1-10)
and 2.8 (1.3-5.1), respectively].
Objectives: The use of asbestos in Sweden was mainly in the
1960ies and import and use of the mineral was banned in the
early mid-seventies, as one of the first countries in the world.
The incidence of mesothelioma in the country started to rise
from very low levels around 1975 and reached around 100 cases a year in 1985, as expected with a latency of 30 years. The
number of cases ought to start sinking in the early 21st century,
especially since strict regulations of asbestos use were in use
already in the mid-1960ies.
Methods: The incidence of pleural mesothelioma for both
sexes cab be seen in the official Swedish publication “Cancer
Incidence in Sweden”, the latest available figures of which is
2012.
Results: Since 1984, a plateau has been reached, with around
100 cases of new pleural mesotheliomas occurring in Sweden
every year, sometimes a little less, more often a little more. No
tendency to declining figures can be seen so far. The median
age at diagnosis has remained the same since the 1960ies, 5470 years. Furthermore, the percentage of women is the same,
15-20 % of the total.
Conclusion: The heaviest exposure took place in Sweden
around 1965; after this the exposure was diminished, but continued until early 1970ies. It has been postulated that 40 years
after exposure, the risk of mesothelioma should decline, but as
stated there are no signs of this in the statistics. The relative
risk for men and women remains about the same, thus it is unlikely that general environmental exposure to asbestos can explain it. Furthermore, the median age at diagnosis has remained
the same. Most of those exposed in the 1960ies should by now
have a fairly advanced age, and thus new age groups must
have had some kind of exposure after 1970. These findings are
disturbing giving the present exposure in many countries.
Keywords: incidence, Sweden, mesothelioma
Conclusion: We found a bidirectional association of pleural
MM with kidney cancer restricted to follow-up within first year,
suggesting increased medical surveillance, while a bidireciMig2016.ORG
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ABSTRACT BOOK
MS07:BAP1 AND GENETICS
MONDAY, MAY 2, 2016
16:30 – 18:00
MS07.01: ASSOCIATION OF BAP1 GENE
EXPRESSION WITH PROTEIN LOCALIZATION AND
SURVIVAL IN EPITHELIOID MPM
Assunta De Rienzo1, Lucian R. Chirieac2, Beow Y. Yeap3 , William
G. Richards1, Raphael Bueno1
Surgery, Brigham and Women’s Hospital and Harvard Medical
School, Boston, MA, UNITED STATES OF AMERICA, 2Pathology,
Brigham and Women’s Hospital and Harvard Medical School,
Boston, MA, UNITED STATES OF AMERICA, 3Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA,
1
Objectives: We have recently demonstrated that diverse
molecular mechanisms may be involved in the tumorigenesis of
malignant pleural mesothelioma (MPM) (Cancer Research, in
press). Unexpectedly, we found that BAP1 gene expression was
associated with significant reduction in survival and was not
correlated with the mutational status of the gene as determined
by sequencing. These observations would seem inconsistent
with its presumed tumor suppressor role. A recent study
documents that frequency of BAP1 mutation is underestimated
by Sanger sequencing which fails to detect large chromosomal
deletions commonly inferred from absence of nuclear BAP1
protein localization by immunohistochemistry (IHC). Here, we
explore BAP1 protein localization by IHC in relation to gene
expression, sex and overall survival (OS).
Methods: One hundred and twenty-eight samples were
selected from the International Mesothelioma Program Tumor
Bank to explore the molecular basis for differential prognosis observed between the two sexes. To minimize competing prognostic influences, the sample set consisted of only
epithelioid MPM samples obtained from equal numbers of male
and female patients who underwent extrapleural pneumonectomy, matched by nodal status and age. Microarray analysis
was performed using 0.25 mg of total RNA and the Ambion
WT Expression Kit. The cRNA was hybridized to Affymetrix®
Human Gene 1.1 ST Array, labeled with GeneChip WT Terminal
Labeling Kit, and then scanned with a GeneAtlas™ Workstation
as recommended by the manufacturer. Quantile normalization
was performed using Bioconductor, and differentially expressed
probes were identified using the LIMMA package. IHC was
performed using BAP1 antibody (Santa Cruz, sc-28383) on a
tissue microarray containing 103 of the 128 samples included
in the microarray analysis. Overall survival was estimated by
the Kaplan-Meier method, with group differences assessed by
the log-rank test. The normalized expression level of BAP1 was
initially grouped into quartiles for exploratory survival analysis.
Patient subgroups with comparable survival were combined for
further analysis in order to detect a meaningful difference using
proportional hazards regression to estimate the hazard ratio
(HR), adjusting for the effect of sex.
Results: After adjusting for the independent effect of sex on
OS (female > male), the highest 3 quartiles of BAP1 expression
were associated with twice the risk of death [HR=2.31; p<0.001]
compared to the lowest quartile, indicating an adverse effect of
elevated BAP1 expression on survival. The survival difference
was similar across sex subgroups (male: HR =2.20; p=0.005;
and female: HR=2.38; p=0.004). BAP1 IHC revealed that only 1
of 27 (4%) cases in the lowest quartile of BAP1 expression exhibited nuclear protein staining, with 21 cases (78%) exhibiting
cytoplasmic staining, and 5 (19%) no staining. By contrast, 17
of 26 (65%) cases in the highest quartile of BAP1 expression
exhibited nuclear staining, 8 (31%) cytoplasmic and 1 (4%) no
staining (p<0.001).
Conclusion: The results suggest that mutations of BAP1,
as indicated by absence of nuclear protein localization, are
associated with lower BAP1 gene expression levels and longer
survival in epithelioid MPM. Further analyses are in progress to
elucidate the clinical correlates of BAP1 status in MPM.
Keywords: BAP1, epithelioid MPM, immunohistochemistry,
suvival
MS07.02: SOMATIC BAP1 AND NF2 MUTATIONS
IN PLEURAL MALIGNANT MESOTHELIOMA
AND THEIR CORRELATION WITH CLINICAL
PHENOTYPES
Spyridon Gennatas1, Shir Kiong Lu1, Hima Anbunathan1, Sanjay
Popat2, Mary E. O’Brien2, Eric Lim3 , Angeles Montero4 , Taqdir
Benepal5, Brendan Tinwell6 , Andrew Nicholson4 , Mark Lathrop7,
Miriam Moffatt1, William O. Cookson1, Anne M. Bowcock1
National Heart And Lung Institute, Imperial College London,
London, UNITED KINGDOM, 2Royal Marsden Hospital NHS
Foundation Trust, London and Surrey, UNITED KINGDOM, 3Royal
Brompton Hospital, London, UNITED KINGDOM, 4Deparatment
Of Histopathology, Royal Brompton and Harefield Hospitals, London, UNITED KINGDOM, 5Department of Oncology, St George’s
Hospital, London, UNITED KINGDOM, 6Department of Histopathology, St George’s Hospital, London, UNITED KINGDOM, 7McGill University and Genome Quebec’s Innovation Centre, Quebec,
AB, CANADA
1
Objectives: We set out to further explore the molecular
alterations underlying pleural MM with the expectation that this
would permit stratification of patients, provide correlations with
clinical information and reveal potential driver mutations that
could provide insights into the development of novel therapeutic
targets.
Methods: We performed whole exome sequencing (WES) on
50 fresh frozen MM tumours (36 epithelioid, 12 biphasic and
2 sarcomatoid). DNA from matched blood was available for 21
of the cases and was also sequenced (Discovery cohort). The
remaining 29 cases were unmatched (Validation Cohort). The
variants were identified with GATK tools and annotated with ANNOVAR. Germline variants, single nucleotide variants (SNV) and
indels with a quality score of less than 50 or those present in
either dbSNP138, 1000 Genomes Project or NHLBI GO Exome
Sequencing Project were filtered out. Potential driver mutations
iMig2016.ORG
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ABSTRACT BOOK
were then identified conditional upon meeting at least one of
the following criteria: 1. SNVs predicted to alter protein function
by Polyphen-2, SIFT and Mutation Taster, 2. Recurrent gene
mutations, 3. Detection of mutation hotspots, 4. Detection of
genes known to be implicated in the tumourogenesis of other
solid tumours. Mutations identified were validated with Sanger
sequencing.
Results: BAP1 and NF2 have been previously described as
genetic drivers of MM. In the discovery cohort we determined
that 19% of tumours (4 of 21) harboured deleterious somatic
mutations in BAP1 and 14% (3 of 21) harboured deleterious somatic mutations in NF2. In the validation set the frequency was
28% (8 of 29) and 14% (4 of 29) respectively. The higher frequency of BAP1 mutations in the validation set is potentially due
to BAP1 mutations of germline origin. We did not see a difference in presence or absence of BAP1 or NF2 mutations and age
at diagnosis of MM. Two tumours had both BAP1 and NF2 mutations and both of these were biphasic. Additionally, we identified a BAP1 mutation in the splice acceptor site of exon 3 in an
MM patient with early onset breast cancer, suggesting that a
germline alteration had contributed to both cancers. A patient
with a deleterious BAP1 mutation (Q684X) from the validation
set had no reported asbestos exposure and had a brother with
lung cancer, a brother with mesothelioma and a brother with
oesophageal cancer. This could expand the spectrum of tissues
implicated in the familial cancer syndrome due toBAP1 mutations. A number of loss-of-function mutations in other important
genes were identified in tumours without BAP1 or NF2 mutations that are being validated and investigated in additional
MMs.
Conclusion: BAP1 and NF2 are the most commonly mutated
genes in MM, but the contribution of germline versus somatic
alterations varies between the two genetic drivers, and a large
percentage of additional drivers remain to be discovered.
Keywords: mesothelioma, BAP1, whole exome sequencing,
genetic drivers
MS07.03: FAMILIES WITH MULTIPLE CASES OF
PLEURAL MALIGNANT MESOTHELIOMA WITHOUT
INHERITANCE OF A BAP1 PREDISPOSING
MUTATION
Valeria Ascoli1, Simona Vatrano2, Luisella Righi2, Ilaria Cozzi1,
Paolo Visca3 , Francesco Facciolo4 , Lucia Rosalba Grillo5, Mauro
Papotti2
genomic status as predisposing risk factor for MM. We report
update data on BAP1 analysis in four families with multiple
cases of MM and asbestos exposure. Preliminary results were
presented at the IMIG Conference 2014, Cape Town.
Methods: Genomic DNA from formalin-fixed paraffin-embedded (FFPE) tissues of one selected case for each family was
obtained (3 epithelioid and 1 biphasic MM), since germline DNA
was not available from peripheral blood. Manual microdissection was performed to separate in all cases the correspondent
normal/neoplastic counterparts, which were analyzed separately. Using Sanger sequencing, the entire sequence of the BAP1
gene was analyzed to screen genetic alterations in coding and
in exonic/intronic junctions. Putative pathogenic variants were
validated via bidirectional re-sequencing of an independent PCR
amplification. Variants were annotated according to the longest
isoform RefSeqs from the Genome Reference Consortium Human Build 37.3 (NM_004656.3) and described according to the
Human Genome Variation Society guidelines. The biphasic MM
was further microdissected in the two neoplastic populations:
epithelioid and sarcomatoid cells. BAP1 immunohistochemistry
(IHC) was performed on FFPE tissues (clone-C4, Santa-Cruz
Biotechnology, CA/USA).
Results: Sequencing analysis of BAP1 gene of tumor samples
of three epithelioid index cases showed no mutations neither in
heterozygosis nor in homozygosis. In the biphasic index case a
non-frameshift InDel was detected in heterozygosis in exon 5
(c.329_335delinsTC) in the epithelioid cells. The same genetic alteration was identified in sarcomatoid cells with a lower
allelic frequency than that seen in epithelioid cells. This BAP1
alteration that led to a shorter BAP1 protein (p.Pro110_Ser111del) has never been reported. Normal tissue from the same
case showed no abnormalities in the BAP1 gene. Therefore, the
identified alteration was proved as a somatic event. For the other three index cases, IHC showed loss of nuclear BAP1 staining,
suggesting other somatic inactivating events, as large deletions
or epigenetic silencing, which could not be identified by Sanger
sequencing (Nasu et al. J Thorac Oncol 2015; Emi et al. J Hum
Genet 2015).
Conclusion: In these four families, there is no evidence of an
inherited mutation of BAP1 by Sanger sequencing. The wild-type
status was proven in tumour tissue (three cases/families) and
in normal tissue (one case/family). Our families with multiple
cases of MM and without inheritance of a predisposing BAP1
mutation are similar to those reported by other investigators
(Popova et al. Am J Hum Genet 2013; Betti et al. Gene Chromosomes Cancer 2014; Sneddon et al. Gene 2015; Cheung et al.
Cancer Genet 2015), suggesting that other genetic or epigenetic
factors may play a role in the high incidence of MM in these
families.
Keywords: BAP1, familial mesothelioma
Radiological, Oncological And Anatomo-pathological Sciences,
Sapienza University, Rome, ITALY, 2Department of Oncology, University of Turin, Turin, ITALY, 3Department of Pathology, Regina
Elena Cancer Institute, Rome, ITALY, 4Department of Oncologic
Thoracic Surgery, Regina Elena Cancer Institute, Rome, ITALY, 5Department of Pathology, San Camillo Hospital, Rome, ITALY
1
Objectives: BAP1 gene germline alterations have been reported in some families with multiple cases of malignant mesothelioma (MM) and very rarely in sporadic MM. Familial clusters
among blood-relatives are ideal candidate to explore BAP1
iMig2016.ORG
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ABSTRACT BOOK
MS07.04: BAP1 AND MESOTHELIOMA
Michele Carbone1, Erin Flores1, Mitsuru Emi1, Todd Johnson2,
Tatsuhiko Tsunoda2, Dusty Behner1, Harriet Hoffman3 , Mary
Hesdorffer4 , Masaki Nasu1, Andrea Napolitano1, Amy Powers1,
Michael Minaai1, Francine Baumann1, Peter Bryant-Greenwood1, Olivia Lauk5, Michaela Kirschner5, Walter Weder5,
Isabelle Opitz5, Harvey I. Pass6 , Giovanni Gaudino1, Sandra
Pastorino1, Haining Yang1
STATES OF AMERICA, 2RIKEN Center for Integrative Medical
Science, Kanagawa, JAPAN, 3Genealogy from the Hart, Honolulu, AL, UNITED STATES OF AMERICA, 4Mesothelioma Applied
Research Foundation, Alexandra, AL, UNITED STATES OF
AMERICA, 5Klinik fur Thoraxchirurgie Universitatsspital, Zurich,
SWITZERLAND, 6NYU Langone Medical Center, New York, AL,
1
Objectives: Germline BAP1 mutations cause a cancer syndrome characterized by high incidence of mesothelioma (MM),
uveal melanoma and other cancers, and by very high penetrance, as all individuals carrying BAP1 mutations developed at
least one, and usually several, malignancies throughout their
lives. Through screening MM patients with histories of multiple
cancers, we studied the prognosis of MM in carriers of germline BAP1 mutations compared to sporadic mesotheliomas, we
investigated the pattern of trasnmission fo these mutations (i.e.,
de novo versus transmission accross multiple generations) and
studied mechanisms of BAP1 carcinogenesis.
Methods: We used a combination of epiedmiological, genealogical studies together with molecular genetics and pathological analyses.
Results: We found that MM in families carrying germline
BAP1 mutations have better prognosis compared to sporadic
mesotheliomas, and we identified four families that shared an
identical BAP1 mutation: they lived across the US and did not
appear to be related. By combining family histories, molecular
genetics, and genealogical approaches, we uncovered a BAP1
cancer syndrome kindred of ~80,000 descendants with a core
of 100 individuals, whose members descend from a couple
born in Germany in the early 1700s who immigrated to North
America. Their descendants spread throughout the country with
mutation carriers affected by multiple malignancies.
Conclusion: Our data show that, once a proband is identified,
extended analyses of these kindreds, using genomic and genealogical studies to identify the most recent common ancestor,
allow investigators to uncover additional branches of the family
that may carry BAP1 mutations. Using this knowledge, we have
identified new branches of this family carrying BAP1 mutations.
We have also implemented early-detection strategies that
help identify cancers at early-stage, when they can be cured
(melanomas) or are more susceptible to therapy (MM and other
malignancies).
MS07.05: BAP-1 CANCER SYNDROME
ASSOCIATED MALIGNANCIES WERE NOT
DETECTED AMONG 558 DANISH PATIENTS WITH
MALIGNANT MESOTHELIOMA
Vasiliki Panou1, Mogens Vyberg2, Christos Meristoudis3 , Oluf D.
Røe4
Department of Respiratory Diseases & Clinical Cancer Research
& Faculty Of Medicine, Aalborg University Hospital & Aalborg
University, Aalborg, DENMARK, 2Institute of Pathology & Clinical
Institute, Aalborg University Hospital & Aalborg University,
Aalborg, DENMARK, 3Institute of Pathology, Aalborg University
Hospital, Aalborg, DENMARK, 4Department of Clinical Medicine,
Aalborg University Hospital, Aalborg, DENMARK
1
Objectives: Recently germline BAP1 mutations were found to
predispose to a cancer syndrome, including malignant mesothelioma (MM), uveal malignant melanoma (UVM), renal cell
carcinoma (RCC) and benign atypical melanocytic tumours
(atypical Spitz tumors [AST]). In the literature there are described families of American and European origin, including a
Danish family, that are reported to be carriers of BAP1 germline
mutations and present with high incidence of MM, UVM, RCC
and AST. Importantly, previous research indicates that MM
patients with BAP1 mutations tend to develop UVM and AST
some years before the MM diagnosis. Hence, it is suggested
that patients with UVM and AST should be tested for BAP1
mutations and if confirmed closely monitored as to early MM
diagnosis. Our aim was to examine whether there was excess of
BAP-1 related tumors in a Danish MM cohort.
Methods: A retrospective analysis of pathology reports of 558
patients diagnosed with MM in pleura, peritoneum, pericardium
and tunica vaginalis in the period 1972-2014 in the Region of
North Jutland, Denmark was performed. Epithelioid, sarcomatoid and biphasic MM subtypes were included in the study population. We investigated whether these patients were previously
or subsequently diagnosed with UVM, RCC or AST.
Results: None of the 558 MM patients had a previous diagnosis of UVM, RCC or AST.
Conclusion: The lack of UVM, RCC and AST tumours in the
large MM study population in North Jutland raises questions
as to how common germline BAP1 mutations that result in both
UVM, RCC, AST and MM appear to be. The current study indicates that the occurrence of UVM, RCC and AST prior or subsequent to a diagnosis of MM is infrequent in Danish patients and
thus, AST´s value as an early MM clinical marker for the general
population is limited. However, research is needed to elucidate
the true incidence of Danish BAP-1 MM patients and their tumor
spectrum.
Keywords: Malignant mesothelioma, BAP1, atypical Spitz
nevus
Keywords: Malignant mesothelioma, BAP1, Cancer Syndrome
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ABSTRACT BOOK
MS08:PATHOLOGY
MONDAY, MAY 2, 2016
16:30 – 18:00
MS08.01: IMMUNOHISTOCHEMICAL DETECTION
OF MTAP OR BAP1 PROTEIN LOSS FOR
MESOTHELIOMA DIAGNOSIS: COMPARISON WITH
P16 FISH
Kazuki Nabeshima1, Tomoyuki Hida1, Makoto Hamasaki1, Shinji
Matsumoto1, Ayuko Sato2, Tohru Tsujimura2, Kunimitsu Kawahara3 , Ainori Iwasaki4 , Yoshinao Oda5
Pathology, Fukuoka University School of Medicine, Fukuoka, JAPAN, 2Pathology, Hyogo College of Medicine, Hyogo, JAPAN, 3Pathology, Osaka Prefectural Medical Center for Respiratory and
Allergic Disease, Habikino, JAPAN, 4Thoracic Surgery, Fukuoka
University School of Medicine, Fukuoka, JAPAN, 5Anatomic Pathology, Kyushu University, Graduate School of Medical Sciences,
Fukuoka, JAPAN
1
Objectives: Objectives. Differentiating malignant pleural mesothelioma (MPM) from reactive mesothelial hyperplasia (RMH)
is sometimes difficult. In this setting, homozygous deletion (HD)
of p16INK4A ( p16; detected using fluorescence in situ hybridization (FISH)) and loss of BAP1 protein expression (detected using
immunohistochemistry (IHC)) are reliable markers for MPM.
Not all laboratories are equipped to perform p16 FISH; IHC is
a more common and feasible technique. The combined loss of
methylthioadenosine phosphorylase (MTAP) and p16 expression
has been proposed as a surrogate marker for p16 HD in pancreatic and lung cancers. We investigated whether detection using
IHC of the loss of expression of the products of genes located in
the 9p21 region ( p14ARF, p15INK4B, p16, and MTAP) could predict
the deletion status detected by p16 FISH in MPM. We also
examined the sensitivity of a combination of IHC of these gene
products and BAP1 for the differentiation of MPM versus RMH,
compared with a combination of BAP1 IHC and p16 FISH.
Methods: Methods. IHC was used to investigate expression
levels of p14ARF, p15INK4B, p16, and MTAP for 43 epithelioid MPM
and 20 RMH. Concordance between their results, including
those for combined expressions, and the deletion status of the
9p21 locus (detected using FISH) was analyzed statistically. We
also examined sensitivities and specificities of combinations of
the IHC expression status of these gene products and BAP1 for
differentiating MPM from RMH, and compared them with those
of BAP1 IHC and p16 FISH.
Results: Results. IHC revealed that loss of expression of p14ARF,
p15INK4B, p16, MTAP and BAP1 occurred in 23.3%, 39.5%,
34.9%, 46.5% and 65.1% of MPM, respectively. p16 HD was
detected in 68.6% of MPM when FISH was used. Among the
four genes, the results of MTAP IHC had the best concordance
with the p16 FISH results (kappa coefficient = 0.65). Moreover,
the loss of p15INK4B -p16 -MTAP revealed using IHC had better
concordance. For predicting p16 HD detected using FISH, loss
of p15INK4B, p16, and MTAP (but not p14ARF) had specificity values
of 100%. The loss of MTAP-BAP1 expression (detected using
IHC) was observed in 85.7% of MPM. The loss of BAP1 using
IHC-p16 HD using FISH was revealed in 94.3% of MPM. Both
combinations had specificity values of 100%.
Conclusion: Conclusion. Combinations of MTAP-BAP1 loss
revealed using IHC can likely detect MPM with good sensitivity
that is higher than those of BAP1 IHC alone or p16 FISH alone.
These combinations could thus serve as useful ancillary IHC for
discrimination of MPM from RMH. A combination of BAP1 IHC
and p16 FISH remains the most accurate ancillary tool.
Keywords: FISH, p16, BAP1, MTAP
MS08.02: UTILITY OF BAP1
IMMUNOHISTOCHEMISTRY AND FISH IN THE
DIFFERENTIAL DIAGNOSIS OF MALIGNANT
MESOTHELIOMA
Kenzo Hiroshima1, Di Wu1, Mizue Hasegawa2, Yasuo Sekine3 ,
Daisuke Ozaki4 , Toshikazu Yusa4 , Zhi Bin Gao5, Yuji Tada6 , Hideaki Shimada7, Masatoshi Tagawa8
1
Pathology, Tokyo Women’s Medical University, Yachiyo,
JAPAN, 2Respirology, Tokyo Women’s Medical University,
Yachiyo, JAPAN, 3Thoracic Surgery, Tokyo Women’s Medical
University, Yachiyo, JAPAN, 4Chiba Rosai Hospital, Ichihara,
JAPAN, 5Yuyao People’s Hospital, Yuyao, CHINA,6Department
of Respirology, Graduate School of Medicine, Chiba University,
Chiba, JAPAN, 7Department of Surgery, School of Medicine,
Toho University, Tokyo, JAPAN, 8 Chiba Cancer Center Research
Institite, Chiba, JAPAN
Objectives: Distinction between malignant mesothelioma
and reactive mesothelial proliferation is sometimes difficult,
especially when the biopsy specimen is small. However, correct
diagnosis is crucial to patient care and compensation. The aim
of this study was to analyze immunohistochemistry (IHC) and
fluorescence in situ hybridization (FISH) of BAP1 in malignant
mesotheliomas and to determine whether they are useful for
the differential diagnosis of malignant mesothelioma.
Methods: IHC and FISH analysis of BAP1 was performed in 28
surgical biopsies from histologically confirmed malignant mesotheliomas. Sixteen were epithelioid mesotheliomas, seven were
biphasic mesotheliomas, and five were sarcomatoid mesotheliomas. We also analyzed surgical biopsies from nine cases with
fibrous pleuritis for control. For FISH analysis, at least 100 cells
were scored for each case. A cut-off value of >15% for homozygous deletion pattern was defined for homozygous deletion.
Results: BAP1 loss by IHC was observed in 56% (9/16) of
epithelioid mesotheliomas, and 57% (4/7) of biphasic mesotheliomas, and none (0/5) of sarcomatoid mesotheliomas.
Homozygous deletion of BAP1 by FISH was observed in 50%
(8/16) of epithelioid mesotheliomas, 14% (1/7) of biphasic
mesotheliomas, and none (0/5) of sarcomatoid mesotheliomas. Heterozygous deletion of BAP1 was observed in 13%
(2/16) of epithelioid mesotheliomas, 71% (5/7) of biphasic
mesotheliomas, and 60% (3/5) of sarcomatoid mesotheliomas.
Abnormality of BAP1 was detected in 81% (13/16) of epithelioid
mesothelioma by BAP1 IHC and/or BAP1 FISH. Abnormality was
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ABSTRACT BOOK
not detected in surgical biopsies from fibrous pleuritis by BAP1
IHC and BAP1 FISH.
Conclusion: Both BAP1 IHC and BAP1 FISH are useful for the
differential diagnosis of epithelioid mesothelioma, although
only BAP1 IHC is useful for the differential diagnosis of biphasic
mesothelioma. Heterozygous deletion of BAP1 is frequently
observed in non-epithelioid mesothelioma and further studies
are needed for clarification of its meaning in malignant mesothelioma.
Keywords: BAP1, mesothelioma, immunohistochemistry, FISH
Conclusion: While reviewing the cases we encountered several
diagnostic issues especially regarding patterns that share many
features (solid and deciduoid, acinar and adenomatoid etc.) as
well as subclassifying sarcomatoid variants. Another issue is the
relative small number of tumor cells in needle biopsies which
makes it harder to evalute and firmly categorize the pattern
into one of the proposed options. A meeting to furtherly define
criteria and recognize specific patterns was needed to improve
the inter-observer agreement and to minimize subjectivity.
Intra-observer agreement was moderate to substantial which
indicates that the proposed classification is quite reproducible
but additional efforts should be probably made to make it an
even more powerful tool.
Keywords: histology, pleural mesothelioma, classification
MS08.03: HISTOLOGICAL EVALUATION OF
MESOTHELIOMAS
Izidor Kern1, Luka Brcic2, Gregor Vlacic1
Pathology, University Clinic Golnik, Golnik, SLOVENIA, 2Pathology, Institute of Pathology, Medical University of Graz, Graz,
AUSTRIA
1
Objectives: Mesothelioma is the most frequent primary
neoplasm affecting the pleura and still has a very grim prognosis. The classification is based on the histomorphological
appearance and distinguishes epitheliod, sarcomatoid and
biphasic variants with the epithelioid type having a longer
overall survival compared to the other two. A new classification
has been proposed by the International mesothelioma working
group. It divides the epithelioid variant into subcategories based
on the histological pattern, namely solid, acinar, adenomatoid,
micropapillary, tubulopapillary, trabecular, small cell, clear
cell, signet ring cell, adenoid cystic, deciduoid, rhabdoid and
pleomorphic. The sarcomatoid variant was subclassified as
conventional (spindle cell), desmoplastic, lymphohistiocytoid
and with heterologous differentiation. Our goal was to assess
inter-observer and intra-observer agreement between two
senior pathologists and one pathology resident applying the
new classification.
Methods: We reviewed the HE slides of 200 consecutive mesotheliomas. There were resection specimens, needle biopsies
(blind and CT guided) and thoracoscopic biopsies from two
different institutions. Following a meeting during which a set
of mesothelioma slides displaying typical patterns for each
histological category was agreed upon and shared among the
pathologists, all the cases were re-evaluated. Fleiss’ kappa was
used to assess inter-observer agreement and Cohen’s kappa to
assess intra-observer agreement.
Results: After the first evaluation the inter-observer agreement
was fair with a kappa value of k=0,372. Following the meeting
the second evaluation round yielded a higher kappa value of
k=0,635 which represents a substantial agreement between
the pathologists. One of the senior pathologists had a substantial intra-observer agreement with a kappa value of k=0,641
while the other senior pathologist and the pathology resident
had a moderate agreement with kappa values of k=0,598 and
k=0,539 respectively.
MS08.04: A GENE PANEL TO DIFFERENTIATE
MALIGNANT PLEURAL MESOTHELIOMAS FROM
BENIGN PLEURAL LESIONS
Rossella Bruno1, Greta Alì2, Riccardo Giannini1, Marco Lucchi3 ,
Franca Melfi4 , Alfredo Mussi3 , Gabriella Fontanini5
Department of Surgical, Medical, Molecular Pathology And
Critical Area, University of pisa, Pisa, ITALY, 2Unit Of Pathological
Anatomy, Azienda Ospedaliero Universitaria, Pisa, ITALY, 3Department of Surgical, Medical, Molecular Pathology And Critical Area,
Division of Thoracic Surgery, University of pisa, Pisa, ITALY, 4Unit
Of Thoracic Surgery, Azienda Ospedaliero Universitaria, Pisa, ITALY, 5Program Of Pleuropulmonary Pathology, Azienda Ospedaliero Universitaria, Pisa, ITALY
1
aggressive and rare tumour associated with asbestos exposure.
The diagnosis of malignant pleural mesothelioma (MPM) is
based on the histological analysis of pleural lesions, however
the morphological separation of benign mesothelial hyperplasia
(MH) from MPM can be exceedingly difficult. As a matter of fact
reactive MH may be extremely florid mimicking mesothelioma.
Nowadays the only robust criterion for malignancy is the presence of mesothelial cells invasion of the chest wall soft tissue
or of the underlying lung parenchyma. Several deregulated
gene pathways have been described in MPM, we investigated
how the over and down expressed genes work together in the
differential diagnosis of MPM and MH. We aimed to assess a
genes panel to be used in the clinical practice by the nCounter
System - NanoString technologies®.
Methods: We designed a custom NanoString Codeset including 113 genes with a crucial role in cancer and 6 reference
genes. The gene expression analysis by the nCounter System
was performed directly, without any amplification steps, on
RNA from 48 formalin-fixed and paraffin-embedded tissues of
epithelioid MPM (32) and MH (16) samples. Raw NanoString
counts for each gene were normalized and statistically processed using the nSolver Analysis Software and the STATISTICA
Software, respectively. To model the gene expression profiles
of MPM and MH samples, both before and after we filtered the
differentially expressed genes, a cluster analysis of expressed
genes was performed using the Euclidean distance between
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ABSTRACT BOOK
samples.
Results: A total of 43 genes resulted deregulated in MPM in
comparison with MH (Mann–Whitney U test; P<.005): 24 genes
exhibited an over expression (EGFR, ITGA3, PAK4, MSLN, GLI2
and others) and 19 showed a down expression (BAP1, ITGA5,
CD44, MMP9, PDGFRB and others). The cluster analysis exposed an evident difference between MPM and MH, particularly
after we filtered the differentially expressed genes.
Conclusion: We analysed a panel of genes, some of which
known as deregulated in MPM, however none of these genes
have yet to be used as a biomarker. To evaluate how transcriptomic data could be applied in the diagnosis of MPM we used
an enzyme-free digital count of mRNA molecules to analyse
simultaneously all the selected genes and to reduce the potential errors associated with multiple qPCR assays. We identified a
specific panel composed of 43 genes, whose expression profile
resulted clearly distinct between MPM and MH. This genes
panel may constitute, after further validation on a larger series
of samples, a powerful tool for the separation of MPM from MH,
improving the current diagnostics methods.
Keywords: Malignant mesothelioma, mesothelial hyperplasia,
differential diagnosis, gene expression profiling
MS08.05: NECROSIS AND SOLID GROWTH
PATTERN AUGMENT NUCLEAR GRADING
IN PREDICTING SURVIVAL IN EPITHELIOID
Alexander J. Gallan1, Viju Ananthanarayanan2, Kenzo Hiroshima3 , Alberto Marchevsky4 , Stephanie Mcgregor1, Angeles Montero5, Kazuki Nabeshima6 , Wickii Vigneswaran7, Ann Walts4 ,
Thomas Krausz1, Aliya Husain1
Department of Pathology, The University of Chicago, Chicago,
IL, UNITED STATES OF AMERICA, 2Loyola University, Maywood,
IL, UNITED STATES OF AMERICA, 3Pathology, Tokyo Women’s
Medical University, Yachiyo, JAPAN, 4Department of Pathology,
Cedars-Sinai Medical Center, Los Angeles, CA, UNITED STATES
OF AMERICA, 5Deparatment Of Histopathology, Royal Brompton
and Harefield Hospitals, London, UNITED KINGDOM, 6Department of Pathology, Fukuoka University, Fukuoka, JAPAN, 7Department of Surgery, The University of Chicago, Chicago, IL, UNITED
STATES OF AMERICA
1
Objectives: A recently described nuclear grading system has
been shown to predict survival in patients with epithelioid diffuse malignant pleural mesothelioma (EMM). The current study
was undertaken to identify additional prognostic characteristics
to augment the nuclear grading system and more accurately
predict survival.
Methods: We analyzed cases of EMM from five institutions
across the United States, England, and Japan from 1998-2013.
Nuclear grade was computed combining nuclear pleomorphism
and mitoses into a grade of I-III using the published system. The
presence or absence of necrosis and patterns of growth were
also evaluated. Overall survival (OS) was used as the primary
endpoint. Data were examined using Student’s t-test.
Results: A total of 117 cases of EMM were analyzed. Of these,
53 (45%) were Grade I, 47 (40%) were Grade II, and 17 (15%)
were Grade III. The mean OS was 28 months. Our multi-institutional data confirmed that higher nuclear grades are associated
with worse OS (Grade I - 40 months, Grade II – 20 months,
Grade III – 13 months, I vs II p<0.001, II vs. III p=0.05, I vs. III
p<0.001). Importantly, Grade II tumors with associated necrosis
behave similarly to Grade III tumors (10 vs 13 months, p=0.37),
and significantly worse than Grade IIs without necrosis (10 vs.
24 months, p=0.003). Additionally, a solid growth pattern was
associated with worse OS (22 vs. 33 months, p=0.02).
Conclusion: This study confirms that nuclear grade predicts
survival in EMM, and identifies the presence of necrosis as a
predictor of especially aggressive behavior in Grade II tumors.
Therefore, we recommend that Grade II tumors with necrosis
be regarded as equivalent to Grade III tumors in behavior.
Additionally, a solid growth pattern, regardless of the nuclear
grade, is associated with a worse overall survival. In conclusion
the assessment of non-nuclear features such as necrosis and
the architectural pattern serve to augment nuclear grading in
predicting survival in patients with EMM.
Keywords: mesothelioma, Nuclear grade, Necrosis
MS08.06: PERITONEAL MESOTHELIOMA:
EVALUATION ON PATHOLOGY REPORTING
Valeria Ascoli1, Ilaria Cozzi2, Giada Minelli3 , Caterina Carnovale
Scalzo2, Emma Rullo1, Elisa Romeo2, Laura Ancona2, Francesco
Forastiere2
Radiological, Oncological And Anatomo-pathological Sciences,
Sapienza University, Rome, ITALY, 2Department of Epidemiology,
Lazio Regional Health Service, Rome, ITALY, 3Unit Of Statistics,
Italy’s Institute of Public Health, Rome, ITALY
1
Objectives: Peritoneal mesothelioma (PeM) has not been
investigated as extensively as pleural mesothelioma. Incidence
rates are low: 0.12/100,000 person-years in men and 0.08 in
women in United States (Am J Epidemiol 2004), and 0.26 in
men and 0.12 in women in Italy, the largest European asbestos
producer (Am J Industr Med 2015). The diagnostic process can
be challenging. Guidelines for the pathological diagnosis have
focused on some site-specific issues. The aim of this study was
to give a contribution to the knowledge of diagnostic practice of
PeM on the basis of pathology reporting.
Methods: The source of pathology reports were a regional
section of the Italian network of Mesothelioma Registries
(Lazio; 2001-2014; 5.5 million people, one tenth of the Italian
population), and a pathology-based archive operating before
the implementation of the registry (1990-2000). We reviewed
our database of 928 mesothelioma cases. Of these, 102 were
PeM (10.3% of all mesotheliomas). We evaluated the report
content and whether and how pathologists follow the immunohistochemistry (IHC) recommendations of the literature
(Husain et al, Arch Pathol Lab Med 2013): a panel of at least 2
mesothelioma markers and 2 carcinoma markers as standard.
All pathology information was manually entered into a specific
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ABSTRACT BOOK
dataset devised for the study. Asbestos exposure was evaluated
through clinical chart review.
Results: Reports consisted of 79 histological diagnoses, 23
cytological diagnoses and 1 autopsy. The mean age at diagnosis
was 68.5 years in women and 63.4 in men; there was a male
predominance (M:F=2.16:1). Available data on asbestos exposure revealed that all persons with ascertained exposure (22%
of 102 PeM) were males. More of 60% of women have unknown
exposure versus 38% of men. The histology ‘subtype unspecified’ was more frequent (n=47; 46%) than epithelioid (n=41;
40%), well-differentiated papillary (n=8), biphasic (n=5) and
sarcomatoid (n=1) mesothelioma. IHC results from 89 reports
(87.3%) showed a wide choice of IHC markers (up to 40 different antibodies, with a mean number for each diagnosis of 5.2).
The most used markers were calretinin (84.3%, pos=98.6%),
HBME-1 (39.3%, pos=97.1), EMA (48.3%, pos=97.7%), pan-cytokeratin (42.6%, pos=97.3%), CEA (41.5%, pos=0%), Ber-EP4
(36%, pos=18.7%) and cytokeratin 7 (37%, pos=90.9%). The
compliance to the IHC standard panel was quite low (16 cases;
18%) with no difference by gender (Pearson’s Chi-squared
P=0.930). Instead, most frequently used panels were those
consisting of at least 2 mesothelioma markers plus 1 carcinoma
marker (26 cases; 29%) and of calretinin-only plus either CEA
or BerEP4 (27 cases; 30%). 55 pathologists were involved, with
a mean number of 5 diagnoses each; 48 pathologists (87%)
reported between 1-2 diagnoses; only 2 pathologists reported
more than 10 diagnoses.
Conclusion: From our small dataset, it emerges an inhomogeneous approach to the diagnosis of PeM, much more than
that observed in pleural mesothelioma. Although the guidelines
are designed to improve the reporting practice, there is a large
scope for improvement in their application, since only a few
pathologists are following them for a standardized diagnosis.
Keywords: pathology reporting, guidelines, immunohistochemistry
MS08.07: THE CRUCIAL CLINICOPATHOLOGICAL
APPROACH IN SUPERFICIAL MESOTHELIAL
PROLIFERATIONS; MESOPATH EXPERIENCE
Francoise Galateau Salle1, Nolwenn Le Stang1, Sylvie Lantuejoul1, Daniel Pissaloux2, Experts Pathologists Mesopath1,
Anabelle Gilg Soit Ilg3 , Patrick Brochard4 , Jean Claude Pairon5,
Arnaud Scherpereel6
with early mesothelioma is crucial for the optimal management
of the patient and also to understand the multistage of carcinogenesis at the dawn of high resolution sequencing. Loss of BAP1
tumor suppressor gene by immunohistochemistry and loss of
CDKN2A ( p16 ) by FISH analysis are useful biomarkers of malignancy working on paraffin embedded tissue biopsy samples. In
this study we aimed to analyze the profiles of such cases and to
propose an algorithm for their clinical management.
Methods: We have selected 78 patients including 40 Atypical
Mesothelial Hyperplasia of undetermined malignancy (AMH), 16
reactive mesothelial hyperplasia (RMH) and 22 mesothelioma
with minimal invasion [MMI], from the MESOPATH files since
1998. Cases included clinicoradiological annotations, survival
and were certified according to the standardized procedure of
certification of the MESOPATH panel. Cases were evaluated
when available for BAP1 loss of expression and P16 by immunohistochemical analysis. FISH studies were performed on
paraffin embedded blocks, with more than 50 nuclei counted. A
cut off value of >20% for homozygous deletion was considered
positive. P16 Heterozygous deletions were excluded. Survival
was evaluated with Kaplan Meier analysis and log rank tests.
Results: We observed a younger mean age in patients with
RMH (55 yrs old range [19; 77]) compared to AMH (68 range
[36;90]) and to MMI (69 range [54;84]) p=0.007 with no significant difference for gender and context of asbestos exposure. BAP 1 loss was seen in 35/73 (37%) of the tissue biopsy
specimens available, and none observed in RMH, while 22
(63%) was observed in AMH, and 13 (59%) in MMI (p<0.001).
Homozygous Deletion of p16 was seen by FISH (22% of cases),
and none in RMH, compared to (18%) in AMH and (43%) in
MMI p<0.004). BAP1 loss and P16 deletion were not mutually
exclusive and both were observed in 6 cases of MMI and 2 cases of AMH. The sensitivity of BAP1 loss by immunohistochemistry and P16 Homozygous deletion by FISH to separate benign
versus malignant proliferation (AMH and MMI) was 8/12 (67%)
with a specificity 27/41 (66%). RMH have a median survival >
to 150 months compared to AMH 31 months and to MMI 15
months. A poorer prognosis was observed in patients with p16
homozygous deletion.
Conclusion: Our results confirmed the 100% specificity of
BAP1 loss and p16 deletion in separating benign versus malignant proliferation. Interestingly in difficult cases of atypical
mesothelial proliferation our study allow to select patients
with superficial malignant proliferation to determine a specific
clinical scheme for the management of the patients.
Keywords: AMH, p16 homoygous deletion, BAP1 loss, clinical
management
Mesopath, Cancer Center Leon Berard, Lyon Cedex ,
FRANCE, 2Biopathology, Cancer Center Leon Berard, Lyon Cedex,
FRANCE, 3Dst, National French Health Institute, Saint Maurice,
FRANCE, 4LSTE-ISPED, Bordeaux Cedex, FRANCE, 5Inserm U 955,
CH CRETEIL, Creteil, FRANCE,6Inserm U774, CHRU Calmette, Lille,
FRANCE
1
Objectives: Separating benign versus malignant proliferation
of the pleura is one of the most challenging situations facing
either the pneumologist or the pathologist. Mesothelioma is a
multistep process of carcinogenesis with a long delay of latency
after asbestos exposure (>40 years). Identification of patient
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ABSTRACT BOOK
MS08.08: MULTI-OMICS INTEGRATION FOR
MALIGNANT PLEURAL MESOTHELIOMA SUBTYPES
CHARACTERIZATION
Yuna Blum1, Annie Renier2, Nabila Elarouci1, Françoise Galateau-Sallé3 , Marie-Christine Copin4 , Paul Hofman5, Fabien
Petel1, Françoise Le Pimpec-Barthes6 , Jessica Zucman-Rossi6 ,
Aurélien De Reyniès1, Marie Claude Jaurand6 , Didier Jean6
Programme Cartes D’identité Des Tumeurs (cit), Ligue Nationale Contre Le Cancer, Paris, FRANCE, 2Inserm U.1162, INSERM
U.1162, PARIS, FRANCE, 3MESOBANK, Lyon, FRANCE, 4CHRU
Lille, Lille, FRANCE, 5CHU Nice, Nice, FRANCE, 6Inserm U.1162,
INSERM U.1162, Paris, FRANCE
1
Objectives: Identification of patient tumor homogeneous
molecular subtypes is essential to understand the underlying
oncogenic scenarii and to develop specific therapies. Malignant
Pleural Mesothelioma (MPM) molecular heterogeneity was
highlighted by previous studies, mainly based on a single omic
technology. We recently defined a robust molecular classification defining two groups (C1 and C2), related to prognosis and
differing by their mutation frequency of the BAP1 tumor suppressor gene and by their engagement in the epithelial-mesenchymal transition (EMT). In this study, we analyzed multi-omics
profiles of MPM with the aim of better characterizing molecular
homogeneous subtypes of MPM.
Methods: The molecular profiles of 50 MPM cultures and 70
frozen MPM tumor samples were obtained for four types of
omics: 1) Transcriptome (Affymetrix/HG-U133-plus-2.0 array),
2) MiRNome (miRNA sequencing), 3) Methylome (Illumina/
Meth450 array), 4) Genome (SNP/Omni-Express-v12.Hg19
array).
sification only based on culture samples. Centroid prediction
showed a high match between the 2 groups C1 and C2 obtained
from the two types of samples. Consistent with our previous
findings, the overall survival rate of patients was lower in C2
than in C1 in both types of samples. Pathway analyses again
identified EMT as deregulated between C1 and C2, and found
new pathways such as angiogenesis, VEGF and BMP signal
pathways. SNP array data showed that the chromosome region
3p21, containing the BAP1 locus, is more frequently deleted in
the C1 group in agreement with the most frequent BAP1 mutations (Fig.1A). Unsupervised clustering based on methylation
profiles classified MPM in groups similar to C1 and C2 both in
culture and tumor samples (Fig.1A). As shown in Fig.1B, significant correlation between gene expression and methylation
status was observed for several EMT biomarkers. MiRNome
data identified several miRNAs deregulated between C1 and C2
groups in the two types of samples. For example, MIR96 was
significantly downregulated in MPM of the C2 group associated
with upregulation of potential target genes, including CAV1, a
gene recently identified as a biomarker of an epithelioid MPM
subgroup of poor prognosis (Fig.1C). Conclusion: Multi-omics integration confirms that MPM
cultures are representative of the heterogeneity of primary
tumors and the occurrence of two main molecular groups in
MPM, linked to genetic and epigenetic changes. This integrative
genomic analysis highlights new biomarkers, pathways and epigenetic regulatory mechanisms specific to each MPM groups.
Keywords: Tumor molecular classification, Signal pathway,
Genomics
Results: Unsupervised consensus clustering based on gene
expression profiles defined two MPM groups both in culture and
tumor samples (Fig.1A) in agreement with our previous clas-
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ABSTRACT BOOK
MS09:SURGERY (TECHINICAL ASPECTS)
14:15 – 15:45
MS09.01: EXTENDED PLEURECTOMY DECORTICATION FOR PLEURAL MESOTHELIOMA
IN THE ELDERLY - THE NEED FOR AN INCLUSIVE
YET SELECTIVE APPROACH
Annabel J. Sharkey, Ricky Vaja, Sara Tenconi, Apostolos Nakas,
David Waller
University Hospitals of Leicester, Leicester, UNITED KINGDOM
Objectives: The median age at diagnosis of patients with
malignant pleural mesothelioma in the United Kingdom is 72
years. In order to continue performing radical surgery we have
employed extended pleurectomy decortication (EPD) in favour
of extrapleural pneumonectomy given the strict selection criteria required. Recent series have shown the feasibility of EPD in
the elderly but with continuing debate about the efficacy of this
treatment we reviewed our experience in order to identify more
detailed selection criteria.
Methods: We reviewed prospectively collected data on all
patients from 1999 to 2015 undergoing EPD with the intent
of achieving macroscopic complete resection. We compared
postoperative outcome and survival in patients 70 years and
over with those younger than 70 years. We correlated clinical
and pathological data with outcome in the two groups.
Results: Seventy nine of 282 patients (28.0%) were 70 years or
over at the time of surgery (median age 65, range 42-81 years)
There were no differences in demographic or pathological
characteristics between the two groups (male; under 70 years,
171 patients (84.2%), 70 years or over, 66 patients (83.5%),
epithelioid; under 70 years, 155 patients (76.4%), 70 years or
over 62 patients (78.5%)). A higher number of patients in the
elderly group required intensive care post-operatively (11 patients (5.4%) vs. 13 patients (16.8%) p=0.004) and developed
atrial fibrillation (29 patients (14.4%) vs. 19 patients (24.7%)
p=0.051). There were no differences in the prevalence of other
post-operative complications between the groups, or in median
length of hospital stay (under 70 years 12 days (range 0-70
days), 70 years or over; 14 days (range 2-93 days) p=0.118).
There was no intergroup difference in in-hospital (3.5 % vs. 6.5
p=0.323), or 90-day mortality (7.9 vs. 10.1% p=0.635). Elderly
patients were less likely to receive adjuvant chemotherapy than
younger patients (75 patients (45.7%) vs. 16 patients (29.6%)
p=0.040) but overall survival was similar; 10.5 months vs. 13.0
months(p=0.683). However, in those node positive patients
survival was significantly decreased in the elderly with non-epithelioid tumours, (3.8 vs. 6.6 months p=0.024) and was also
decreased in the elderly with epithelioid disease (9.6 vs. 13.5
months, p=0.485). Survival was similar in all node negative
patients. On multivariate analysis, age was not a significant
prognostic factor, although lack of adjuvant therapy (HR 2.088
95%CI 1.372-3.176 p=0.001) and pre-operative anaemia (HR
1.976 95%CI 1.294-3.017 p=0.002) remained poor prognostic
factors.
Conclusion: Whilst age in isolation should not be an exclusion
criterion for EPD for pleural mesothelioma, in the elderly a more
rigorous preoperative evaluation of nodal disease and an additional assessment of fitness for adjuvant chemotherapy or the
consideration of neoadjuvant therapy are recommended.
MS09.02: A NEW PROGNOSTIC SCORE FOR
TREATMENT ALLOCATION FOR MULTIMODALITY
THERAPY FOR MALIGNANT PLEURAL
MESOTHELIOMA - AN UPDATE
Isabelle Opitz1, Martina Friess1, Olivia Lauk1, Thomas Frauenfelder2, Thi Dan Linh Nguyen-Kim2, Ilhan Inci1, Sven Hillinger1,
Didier Schneiter1, Burkhardt Seifert3 , Rolf Stahel4 , Walter
Weder1
SWITZERLAND, 2Institute of Diagnostic And Interventional Radiology, University Hospital Zurich, Zurich, SWITZERLAND, 3Department of Biostatistics, Epidemiology, Biostatistics And Prevention
Unit, University of Zurich, Zurich, SWITZERLAND, 4Clinic For
Oncology, University Hospital Zurich, Zurich, SWITZERLAND
1
Objectives: We developed a Multimodality Prognostic Score
(MMPS) in our patient cohort receiving induction chemotherapy
followed by extrapleural pneumonectomy (EPP) or pleurectomy/
decortication (P/D) to facilitate the decision for surgery after
induction chemotherapy.
Methods: A 4 variable MMPS was developed including
pre-chemotherapy tumor volume (>500 ml), progressive
disease (PD) after induction chemotherapy (according to
modified RECIST criteria), pre-chemotherapy CRP (> 30mg/ml)
and non-epithelioid histological subtype. Overall survival (OS)
was calculated from the first cycle of induction chemotherapy
until death, and association with the score was analyzed using
Kaplan-Meier curve and log rank test.
Results: Between 1999 and 2015, 253 patients were intended to be treated with induction chemotherapy plus EPP. In 63
undergoing EPP and 20 undergoing pleurectomy/decortication
(P/D) all variables of MMPS were available. Median age at diagnosis was 61 years in the EPP group and 65 in the P/D group.
Epithelioid type was diagnosed in 81% of the EPP and 95% of
the P/D group. IMIG stage III in EPP group was 63% and 65% in
the P/D group. In the EPP cohort patients with score 0 survived
significantly longer than patients with score 3 or higher (Figure
1). The median OS for patients of the EPP cohort was 34 months
(95% CI, 18-50) for score 0, 15 months (9-21) for score 1, 12
months (8-16) for score 2 and 4 months (3-6) for score 3 and 4.
In the P/D group the maximum score reached was 2 in only one
patient. All the others had a score of 0 or 1. The median OS for
score 1 was 30 months (95%CI: 25-36) and 17 months for score
2, but 70% percent of the cases were censored. iMig2016.ORG
44
ABSTRACT BOOK
Results: Patients undergoing salvage EPD had similar survival
to those undergoing more recognized regimes of planned surgery before or after chemotherapy and had better survival than
those patients in whom chemotherapy was reserved for disease
progression after surgery. This was even though the salvage
group were more likely to have nodal metastases. Sal- Neoadvage juvant
n=29 n=18
Adjuvant
n=67
Reserved
for rep
currence
n=50
Median
survival from
diagnosis
(months)
22.6
25.5
22.2
14.6
tion chemotherapy followed by EPP with different MMPS.
Cycles of
chemotherapy
4 (212)
4.5 (2-7)
4 (1-10) 5 (1-7)
Conclusion: Our Multimodality Prognostic Score considering clinical variables already available before surgery allows
identification of mesothelioma patients who would not get any
relevant benefit from an intensified therapy. The concept is
currently under prospective evaluation
Age (years)
61
(4577)
61 (5077)
63 (4276 (52-80) 0.06
80)
Male
gender (%)
86.2
66.7
92.5
82.0
0.035
Epithelioid
cell type
(%)
72.4
66.7
86.6
80
0.272
Node
positive
(%)
74.2
47.1
67.8
46.7
0.035
0
4.5
0
II 6.9
33.3
15.2
22
III 62.1
44.4
62.1
62.0
IV 27.6
22.2
18.2
16.0
Figure 1: Median OS and 95% CI of patients undergoing induc-
Keywords: multimodality treatment, prognosis, Macroscopic
Complete Resection
MS09.04: IT’S NEVER TOO LATE TO OPERATE
- AN ANALYSIS OF SALVAGE SURGERY
FOR PROGRESSING MALIGNANT PLEURAL
MESOTHELIOMA
Annabel J. Sharkey, Rocco Bilancia, Ricky Vaja, Sara Tenconi,
Apostolos Nakas, David Waller
University Hospitals of Leicester, Leicester, UNITED KINGDOM
Objectives: The use of extended pleurectomy decortication
(EPD) as part of multimodality therapy for malignant pleural
mesothelioma remains debatable and many patients receive
chemotherapy primarily. We have been asked to consider EPD
as an afterthought following the failure of primary oncological
treatment, therefore we aimed to determine whether there is a
benefit in performing a potentially morbid operation in those
thought to have a poor prognosis.
Methods: From a prospective database we analyzed 184
patients undergoing EPD as part of multimodality therapy. The
clinicopathological data and outcome of patients undergoing
salvage surgery for disease progression after initial chemotherapy were compared with the 3 other main therapeutic strategies: neoadjuvant chemotherapy, adjuvant chemotherapy and
chemotherapy reserved for disease progression after EPD. All
patients underwent similar preoperative staging with CT but
without routine mediastinoscopy or CTPET.
IMIG stage I 3.4
0.007
0.873
0.325
Conclusion: Lung sparing radical surgery for malignant pleural
mesothelioma should not necessarily be denied to patients
who have undergone first line chemotherapy and in whom their
disease has progressed but remains resectable.
MS09.06: EVOLUTION OF SURGICAL APPROACH
IN MALIGNANT PLEURAL MESOTHELIOMA
Seiki Hasegawa1, Toru Nakamichi1, Ayumi Kuroda2, Masaki
Hashimoto1, Teruhisa Takuwa1, Seiji Matsumoto1, Yoshitomo
Okumura3 , Nobuyuki Kondo1, Fumihiro Tanaka4 , Noriaki Tsubota5, Norihiko Kamikonya6 , Tohru Tsujimura7, Takashi Nakano8
Thoracic Surgery, Hyogo College of Medicine, Nishinomiya,
JAPAN, 2Department of Thoracic Surgery, Hyogo College of Medicine, Nishinomiya, JAPAN, 3Thoracic Surgery, Itami City Hospital,
1
iMig2016.ORG
45
ABSTRACT BOOK
Itami, JAPAN, 4Surgery Ii, University of Occupational and Environmental Health, Kitakyusyu, JAPAN,5Thoracic Oncology, Hyogo
College of Medicine, Nishinomiya, JAPAN, 6Radiation Oncology,
Hyogo College of Medicine, Nishinomiya, JAPAN, 7Department
of Pathology, Hyogo College of Medicine, Nishinomiya, Hyogo,
JAPAN, 8Respiratory Medicine, Hyogo College of Medicine, Nishinomiya Hyogo, JAPAN
Conclusion: Less invasive surgery for MPM yielded lower
surgical risk and also comparable or better survival.
Keywords: extrapleural pneumonectomy, Surgery, pleurectomy/decortication, mesothelioma
Objectives: The aim of this study was to verify whether or not
our approach for less invasive surgery for malignant pleural
mesothelioma (MPM) yielded lower surgical risks as well as
comparable survival in comparison with highly invasive surgery.
Methods: We retrospectively reviewed all patients in a prospective database of MPM Surgery Program at Hyogo College
of Medicine between July 2004 and December 2015. Patients
with histologically confirmed resectable MPM, IMIG cT13N0-1M0 disease, PS 0-1, no major comorbidity, and written
informed consent were registered. All the patients underwent
multimodality treatment with neoadjuvant/adjuvant chemotherapy, surgery with or without 54Gy hemithoracic radiation
therapy. Group 1; Patients underwent trimodality treatment with
conventional extrapleural pneumonectomy (EPP) until August
2009. Group 2: Patients underwent trimodality treatment with
less invasive EPP, using hybrid VATS/open technique, since September 2009 to date. Group 3: Patients underwent bimodality
treatment with pleurectomy/decortication (P/D) and neoadjuvant plus adjuvant chemotherapy as far as macroscopic complete resection could be achieved since September 2012. All the
patients were followed every three months after discharge until
death. There was no censored case.
Results: Of a total of 129 patients registered, 104 (81%)
completed surgery: Group 1 (n=26), Group 2 (n=34), and Group
3 (n=44). Patient characteristics and the results are shown in
Table 1. Macroscopic complete resection was achieved in 95%
(99/104) of patients, and 30-/90-day mortality rates were 1.9%
(2/104) and 3.9% (4/104), respectively. 2-year survival rate and
median survival time of Group 1/2/3 were 38%/75%/77%, and
17.7m/43.3m/not reached, respectively (Figure 1).
MS10:NOVEL TARGETS ENTERING IN THE CLINIC
14:15 – 15:45
MS10.01: PHASE I STUDY OF ANTI-MESOTHELIN
ANTIBODY DRUG CONJUGATE ANETUMAB
RAVTANSINE IN PATIENTS WITH METASTATIC
MESOTHELIOMA
Raffit Hassan1, Johanna C. Bendell2, George R. Blumenschein,
Jr3 , Hedy Kindler4 , Kathleen N. Moore5, Alessandro D. Santin6 ,
Shelly M. Seward7, John Nemunaitis8 , Prabhu Rajagopalan9,
Annette Walter10, Nenad Sarapa9
Thoracic And Gastrointestinal Oncology Branch, National Cancer
Institute, Bethesda, MD, UNITED STATES OF AMERICA, 2Drug
Development Unit, Sarah Cannon Research Institute, Nashville,
TN, UNITED STATES OF AMERICA, 3The University of Texas MD
Anderson Cancer Center, Houston, TX, UNITED STATES OF
AMERICA, 4Radiology, University of Chicago Hospital, Chicago,
IL, UNITED STATES OF AMERICA, 5University of Oklahoma Health
Sciences Center, Oklahoma City, OK, UNITED STATES OF AMERICA, 6Yale Cancer Center, New Haven, CT, UNITED STATES OF
AMERICA, 7Karmanos Cancer Center, Detroit, MI, UNITED STATES
OF AMERICA, 88. Mary Crowley Medical Research Center, Dallas,
TX, UNITED STATES OF AMERICA, 9Bayer HealthCare Pharmaceuticals, Whippany, NJ, UNITED STATES OF AMERICA, 10Bayer
Pharma AG, Berlin, GERMANY
1
Objectives: Anetumab ravtansine (BAY 94-9343; AR) is a novel
fully humanized anti-mesothelin IgG1 antibody conjugated to a
ravtansine, a maytansine derivative DM4 antitubulin cytotoxic
agent. A phase I study evaluating the safety, PK and tumor response with AR was conducted in patients (pts) with advanced
solid tumors (NCT01439152). We report here results with q3w
dosing in patients with mesothelioma.
Methods: AR was administered IV every 21 days (q3w) in 77
pts: 45 pts in 10 dose escalation cohorts from 0.15 to 7.5 mg/
kg (21 mesothelioma, 9 pancreatic, 5 breast, 4 ovarian, 6 other),
and 32 pts in 2 expansion cohorts (12 mesothelioma and 20
ovarian); 38 pts were treated at MTD in escalation and expansion cohorts (16 mesothelioma, 21 ovarian, 1 breast). Clinical
and laboratory safety assessments were made on D1, D8 and
D15 in C1-C3 and on D1 in subsequent cycles. Tumor assessments were made q6wks up to C8 and q12wks thereafter.
Mesothelin expression in archival tumor samples was assessed
retrospecively by IHC (SP74, Ventana).
Results: A total of 77 pts (45 females) were treated with AR
q3w and evaluable for safety; mean age 62 yrs (range, 18-84
iMig2016.ORG
46
ABSTRACT BOOK
yrs), body weight 77 kg (44-113 kg), ECOG ≤1, median prior cytotoxic regimens: overall 4 (1-9), mesothelioma 1 (1-4). The MTD
for AR was 6.5 mg/kg q3w (one DLT: G3 AST increase); DLTs at
7.5 mg/kg indcluded 1 pt with G2 keratitis and G3 neuropathy,
and 1 pt with G4 keratitis and G2 neuropathy. Only one DLT (G3
hyponatremia, 5.5 mg/kg) and markedly fewer AEs occurred at
doses below the MTD. No drug-related deaths and few drug-related SAEs (7 total and 5 at MTD) were reported. Seventeen of
38 (45%) pts in total or 7 of 16 (44%) mesothelioma pts at MTD
had drug-related AE requiring dose reduction (G1-4 keratitis,
G2-3 neuropathy, G3 fatigue, anorexia, asthenia, diarrhea, N&V,
AST increase). LFT increases were the most common drug-related laboratory abnormality at MTD: AST in 7 pts (2 G3), ALT
in 6 pts (no G3), alkaline phosphatase in 4 pts (one G3) and
bilirubin increase in 1 pt (no G3). There were no drug-related G3
hematological abnormalities at any dose. Fourteen of 38 (37%)
pts total or 4 of 16 (25%) mesothelioma pts at MTD had G1-4
keratitis (worst G3-4 in 3 pts, blurred vision in 10, dose reduction in 8, dose delay in 11, all fully reversible). Five of 16 (31%)
mesothelioma pts treated with AR at the MTD had a durable
partial response (PR; >600 days in 4 pts) and 7 (44%) had stable disease. The five PRs occurred in 10 mesothelioma pts who
received AR as second line treatment (50% response rate).
Conclusion: AR at the MTD (6.5 mg/kg) showed encouraging efficacy with durable PRs in pts with metastatic mesothelioma. At the MTD, all drug-related AEs were reversible,
non-life-threatening and manageable by dose modification.
Given this benefit-risk ratio, the recommended phase II dose of
AR in second line treatment of advanced mesothelioma is 6.5
mg/kg IV q3w.
Keywords: antibody drug conjugate, mesothelioma, mesothelin
MS10.02: PHASE 1 DOSE EXPANSION
EXPERIENCE OF ADI-PEG20, PEMETREXED AND
CISPLATIN IN PATIENTS WITH MALIGNANT
MESOTHELIOMA (TRAP STUDY)
Peter Szlosarek1, James Spicer2, Melissa Phillips3 , Jeremy
Steele3 , Hannah Rush4 , Monica Diaz5, Adalberto Barba5, Amanda Johnston5, Ramsay Khadeir1, Michael Sheaff3 , John Bomalaski5, Simon Pacey6
Centre For Molecular Oncology, Barts Cancer Institute, London,
UNITED KINGDOM, 2King’s College London, London, UNITED
KINGDOM, 3Medical Oncology, St. Bartholomew’s Hospital,
London, UNITED KINGDOM, 4Guy’s & St Thomas’ NHS Foundation Trust, London, UNITED KINGDOM, 5Polaris Pharma Inc., San
Diego, AL, UNITED STATES OF AMERICA, 6Oncology, University of
Cambridge, Cambridge, UNITED KINGDOM
1
chemotherapy that included patients with ASS1-deficient
malignant pleural mesothelioma (MPM) and observed a 78%
partial response (PR) rate in the dose-escalation portion of the
study (AACR-NCI-EORTC, Boston 2015). Here, we describe our
dose-expansion experience at the maximum tolerated dose
(MTD) in patients with MPM.
Methods: Main inclusion criteria were: ≥ 18 years, PS ≤ 1,
tumour ASS1 loss (≤ 50% ASS expression by IHC) with adequate organ function and written, informed consent. Patients
were excluded with: symptomatic CNS metastases, significant
concurrent morbidity, therapeutic anticoagulation, history of
seizures, or allergy to trial medication(s). Patients were treated
at the MTD as follows: weekly ADI-PEG 20 (36 mg/m2 IM) plus
pemetrexed 500 mg/m2 and cisplatin 75 mg/m2 both given
every 3 weeks. ADI-PEG 20 alone was allowed after 18 weeks
if there was stable disease (SD) or better. Adverse events were
graded using CTCAE v4.03. Radiological disease response was
assessed every 6 weeks by modified RECIST and peripheral
blood samples were collected to measure plasma arginine and
citrulline levels and antibodies to ADI-PEG 20.
Results: 82 patients were screened for MPM ASS1 expression, 32 (39%) were ASS1-deficient and 20 (62.5%) were
enrolled. Subsequently, 17 patients were eligible for toxicity
and response assessment, that included 5 patients from the
dose-escalation cohort. Demographics - 16 M:1 F, Age range
60-82, Subtype (epithelioid 4, biphasic 6, sarcomatoid 7). Grade
3 or higher AEs related to pemetrexed and cisplatin were (6
pt): nausea/vomiting (2), neutropenia (2), others 3 (number of
events 6); 16 AEs were reported possibly related to ADI-PEG 20.
Mean cycles (weeks) of treatment: 22.0 (range 5.7-44.3 weeks).
Mean (weeks) arginine depletion is 9.6 (range 3-18) weeks in all
treated patients. Response by modified RECIST was as follows:
53% (9/17) had a PR and 47% (8/17) had SD for a disease
control rate (DCR) of 100% (17/17).
Conclusion: The triplet combination of ADI-PEG 20+Pemetrexed+Cisplatin was well tolerated as described in the earlier
dose-escalation portion. Robust clinical activity has been observed with a 100% DCR in a population enriched for biphasic
and sarcomatoid pathology. The tolerability and high response
rate in the poorer prognosis patients suggest that this combination may have clinical utility as first line treatment for ASS1-deficient MPM. The expansion-phase study is ongoing to recruit
30 patients in total and a global randomized placebo-controlled
P2/3 study is planned in patients with ASS1-deficient mesothelioma.
Keywords: personalized therapy, mesothelioma, ADI-PEG20,
ASS1
Objectives: Loss of the metabolic tumor suppressor, argininosuccinate synthetase (ASS1), results in an aggressive phenotype but sensitizes malignant mesothelioma cells to apoptosis
with the arginine depleting agent, pegylated arginine deiminase
(ADI-PEG 20) which potentiates the cytotoxic effect of pemetrexed. Thus, we initiated a phase I study (NCT02029690) of
ADI-PEG 20 combined with first-line pemetrexed and cisplatin
iMig2016.ORG
47
ABSTRACT BOOK
MS10.03: TREMELIMUMAB AND DURVALUMAB
COMBINATION FOR FIRST AND SECOND-LINE
TREATMENT OF MESOTHELIOMA PATIENTS: THE
NIBIT-MESO-1 STUDY
MS10.04: PHASE 2 NEOADJUVANT STUDY OF
VS-6063, A FAK INHIBITOR, IN SUBJECTS WITH
SURGICALLY RESECTABLE MALIGNANT PLEURAL
MESOTHELIOMA
Luana Calabro’1, Aldo Morra2, Diana Giannarelli3 , Diego Annesi1, Erica Bertocci1, Riccardo Danielli1, Maresa Altomonte1,
Anna Maria Di Giacomo1, Michele Maio1
Raphael Bueno1, William Richards1, Ritu Gill1, Julianne Barlow1,
Patrick Lizotte2, Kwok Wong2, Mark Bittinger2, Yan Wang3 , Lou
Vaickus3 , David T. Weaver3
1
Medical Oncology And Immunotherapy Division, University Hospital of Siena, Siena, ITALY, 2Euganea Medica Radiology Center,
Padua, ITALY, 3Regina Eelena National Cancer Institute, Rome,
ITALY
1
Objectives: Malignant mesothelioma (MM) has a very dismal
prognosis and treatment of MM patients remains largely unsatisfactory highlighting the need for new therapeutic approaches.
The anti-CTLA-4 monoclonal antibody (mAb) tremelimumab has
shown promising activity in pre-treated MM patients: disease
control rate (DCR) was 31%, and survival rate at 1- and 2-years
was 48.3% and 36.7%, respectively (Calabrò et al., Lancet
Oncol, 2013). These initial findings were corroborated by a
second study in which, based on pharmacokinetic analyses, an
intensified schedule of tremelimumab was utilized. Fifty-two %
of patients achieved a DCR (median duration 10.9 months) (Calabrò et al., Lancet Respir Med, 2015). These intriguing clinical
results and the emerging efficacy of immunomodulatory mAb
targeting the PD-1/PD-L1 axis in different tumor types, prompted us to design the NIBIT-MESO-1 study aimed to investigate
the efficacy of tremelimumab combined with the anti-PD-L1
durvalumab (MEDI4736) in MM patients.
Objectives: Malignant pleural mesothelioma (MPM) is an aggressive tumor in the pleural lining of the lung with early recurrence and a mortality rate of 95-99%. VS-6063 is an oral Focal
Adhesion Kinase (FAK) inhibitor that prevents integrin-mediated
activation of multiple downstream signal transduction pathways
inhibiting tumor cell migration, proliferation and survival in
MPM. The primary objectives of this study are to identify biomarkers associated with response to VS-6063, and to establish
target inhibition in tumor. Secondary endpoints include safety,
pharmacokinetics, and objective response to VS-6063.
Methods: Trial Design: The NIBIT-MESO-1 trial is a phase
II, open-label, study that will enroll 40 unresectable, first-or
second-line pleural or peritoneal MM patients with ECOG performance status 0 or 1. Patients will receive tremelimumab at
1 mg/Kg i.v. every 4 weeks (Q4W) for 4 doses, and durvalumab
at 20 mg/Kg i.v. Q4W for 12 months. Patients with progressive disease during the first 12 months of treatment or in the
follow-up phase may be retreated with the combination of the
two drugs. Modified RECIST (Byrne et al., Ann Oncol, 2004) and
RECIST 1.1 will be utilized to assess tumor responses in pleural
and peritoneal MM, respectively. Primary objective is immune-related (ir)-objective response rate; secondary objectives
are ir-DCR; ir-progression free-survival (PFS), overall survival,
DCR, PFS, and safety. Efficacy secondary endpoints will be
explored per PD-L1 expression on tumor tissues. Clinical results
will be correlated with extensive phenotypic, functional and humoral studies (ClinicalTrials.gov Id: NCT02588131). The study is
actively recruiting and 10 patients have been so far enrolled.
Results: From October 2015 to December 2015, 10 MM patients (9 pleural, and 1 peritoneal), median age 64 years (range
45-77), M/F= 8/2, have been enrolled. Mesothelioma histology
was epithelioid (n=9) or undefined (n=1). Patients received a
median of 2.5 doses of therapy (range= 1 to 3) in first (2 patients) or second line (8 patients), and are all on treatment. No
grade 3-4 treatment-adverse events have been observed so far.
A preliminary safety analysis is ongoing.
Conclusion: The study is in progress and actively recruiting
Keywords: tremelimumab, mesothelioma, durvalumab, immunomodulatory monoclonal antibody
Brigham and Womens Hospital, Boston, MA, UNITED STATES OF
AMERICA, 2Belfer Center Dana Farber Cancer Institute, Boston,
MA, UNITED STATES OF AMERICA, 3Verastem, Inc., Needham,
Methods: An open label, single center, neoadjuvant window of
opportunity study design was incorporated to monitor biomarker changes from tumor biopsies and plasma in subjects with
MPM who are eligible for extirpative surgery. Approximately
19 subjects received VS-6063 400mg BID for 12 days (Cohort
1) or 35 days (Cohort 2). Tumor volume measurements were
calculated from PET-CT. Definitive surgery occurred after a 30
day follow-up period for Cohort 1 and after 7 days for Cohort 2.
Analysis of tumor immunomodulation was investigated by comparing tumor specimens collected at diagnosis/screening, on
drug treatment at Day 12 or Day 35, and at surgery. Likewise,
blood samples were collected at Day 0 and Day 12 to investigate circulating biomarkers.
Results: Four of the nineteen patients from Cohort 1 and
Cohort 2 had tumor volume reduction of 30% or greater during
neoadjuvant VS-6063 treatment. Surgical specimens from
Cohort 2 patients and mesothelioma patients not receiving
VS-6063 treatment were compared by multi-parameter flow
cytometry with immune biomarkers. Whereas tumor CD3+
T cell infiltrates were observed over a wide range, the T, B,
myeloid populations were not statistically different between
these groups. However, the CD123+ plasmacytoid dendritic
cells were decreased (p = 0.03) in VS-6063-treated patients.
Comparing the specimens isolated before and after VS-6063
treatment, two patients showed increased CD8+ T cells, and
one of the patients showed 40% fewer FOXP3+ T cells (Tregs)
by IHC. Higher baseline FOXP3+ T cells were an indicator of
tumor reduction in the Cohort 2 patients. Plasma cytokine
changes were monitored by comparing blood samples isolated
at screening with Day 12 of VS-6063 treatment. Notably, IL-10,
an immunosuppressive cytokine was significantly diminished
after VS-6063 (p = 0.0186).
Conclusion: Neoadjuvant treatment of mesothelioma patients
with VS-6063 for either 12 or 35 days was associated with a tumor volume reduction and tumor immunomodulation. As IL-10
is an immunosuppressive cytokine produced by FOXP3+ T cells
the decreased circulating IL-10 levels are potentially indicative
iMig2016.ORG
48
ABSTRACT BOOK
of tumor immunomodulation by VS-6063. Further evaluation of
biomarker responses to VS-6063 in the neoadjuvant setting is
warranted.
Keywords: FAK, immunomodulation, neoadjuvant, mesothelioma
MS10.05: PHASE I EXPERIENCE WITH TARGOMIRS
Nico Van Zandwijk1, Nick Pavlakis2, Steven Kao3 , Stephen
Clarke4 , Anthony Linton5, Michael Boyer3 , Himanshu Brahmbhatt6 , Jennifer Mcdiarmid6 , Yennie Huynh7, Felicity Leslie7,
Helen Foster6 , Scott Pattison6 , Glen Reid7
Northern Cancer Institute, Asbestos Diseases Research Institute,
Concord, NSW, AUSTRALIA, 2Northern Cancer Institute, Sydney,
NSW, AUSTRALIA, 3Chris O’Brien Lifehouse, Camperdown, NSW,
AUSTRALIA, 4Northern Cancer Institute, University of Sydney,
Sydney, NSW, AUSTRALIA,5Dept Oncology Concord Hospital,
Concord, NSW, AUSTRALIA, 6EnGeneIC, Lane Cove, NSW, AUSTRALIA, 7Asbestos Diseases Research Institute, Concord, NSW,
AUSTRALIA
than 40 weeks. Repeat imaging revealed 9 patients with SD at 8
weeks (modified RECIST). Updated results will be presented.
Conclusion: Weekly infusions of 5 billion TargomiRs are rather
well tolerated. Rapidly transient inflammatory (cytokine-mediated) symptoms were noticed shortly after the infusions. Blood
examinations reveal a dynamic pattern of changes in heamatologic and non-heamatologic parameters after TargomiR infusion. It is assumed that transient hypophosphatemia is induced
by the cytokine reactions elicited by TargomiRs. The transient
subtle ST-T changes noted at repeat electrocardiography are
also thought to be associated with the inflammatory syndrome.
Documentation of 2 objective responses and 9 patients with
stable tumour measurements after 8 weeks of TargomiR treatment point to single agent activity of TargomiRs.
Keywords: phase I study, microRNA-based therapy, Inflammatory reactions, Malignant pleural mesothelioma
1
Objectives: MesomiR 1 is the first-in-man phase I study
testing the intravenous administration of TargomiRs. TargomiRs
are nanoncells (EDVTM) packaged with miR-15/16- derived
microRNA mimics targeted with EGFR antibodies. Patients with
malignant pleural mesothelioma (MPM) recurring after standard therapy were asked to participate.
Methods: A 3-6 patient dose escalation cohort design
examining weekly/twice weekly TargomiR infusions is being used (Clinical Trials.gov: NCT02369198/ ANZCTR-ACTRN12614001248651). Patients tolerating TargomiR infusions
well were allowed to continue protocol therapy for at least 8
weeks. Fifty percent of the Maximal Tolerated Dose (MTD)
previously established for EDVs was chosen as a first dose level
(= 5 billion TargomiRs packaged with a total of 1.5 microgram
miR-15/16 mimics). CT, FDG-PET and pulmonary function
assessment were scheduled at 8 week intervals and Quality-of-Life (QoL) questionnaires (EORTC) were requested on a
weekly basis.
Results: Fifteen male and three female MPM patients, failing
on standard therapy received different doses and schedules
of TargomiRs ranging from 1 billion (once a week) to 5 billion
(twice a week). Currently a total of 221 weeks of TargomiR
treatment is being analysed. TargomiRs at a dose of 5 billion
were rather well tolerated when administered once a week.
Rapidly transient inflammatory symptoms including shivering/
rigor, temperature elevation and pain at tumour sites in the
chest were noted shortly after TargomiR infusion but seldomly
exceeded CTC grade 3. The transient inflammatory symptoms
were accompanied by neutrophilia, lymphopenia, hypophosphatemia, and sometimes elevation of liver enzymes. Transient
(asymptomatic) ECG (ST-T) changes were noted in 4 patients
and were also suspected to be part of the inflammatory reactions noted. Repeat imaging revealed 2 objective responses (PR)
and both patients remained on experimental therapy for more
MS10.06: ONCOLYTIC HERPESVIRUS THERAPY
FOR MESOTHELIOMA - A PHASE I/IIA TRIAL OF
INTRAPLEURAL HSV1716 (NCT01721018)
Penella Woll1, Sarah Danson1, John Edwards2, Patricia Fisher1, Kevin Blyth3 , Joe Conner4
University of Sheffield, Sheffield Teaching Hospitals, Sheffield,
UNITED KINGDOM, 2Northern General Hospital, Sheffield, UNITED KINGDOM, 3Respiratory Medicine, Queen Elizabeth University
Hospital, Glasgow, UNITED KINGDOM, 4Virttu Biologics Ltd,
Glasgow, UNITED KINGDOM
1
Objectives: Malignant pleural mesothelioma (MPM) remains
a major challenge, with limited therapeutic options. Multifocal
intrapleural disease can cause disabling symptoms of pain and
breathlessness, in the absence of distant metastases, so an
intrapleural treatment approach is attractive. HSV1716 (Seprehvir) is a mutant herpes simplex virus type 1 deleted in the RL1
gene which encodes the protein ICP34.5, a specific determinant of virulence. Mutants lacking the RL1 gene are capable
of replication in actively dividing cells but not in terminally
differentiated cells – a phenotype exploited to selectively target
and kill tumour cells. Additionally, oncolysis of the target cancer
cell also stimulates an anti-tumour T-cell mediated immune
response Activity against mesothelioma has been demonstrated in animal models. Clinical studies with HSV1716 have
been completed in adult glioma, melanoma, H&NSCC and it is
well-tolerated with no shedding in patients. We have therefore
designed and implemented a phase I/IIa trial to determine the
safety and potential for efficacy of HSV1716 given intrapleurally
to patients with MPM.
Methods: The study is an open label, dose escalation, phase
I/IIa trial currently open at two clinical centres. Patients with
a histological diagnosis of MPM and a Rocket® or PleurX®
indwelling pleural catheter (IPC) are eligible if they have performance status ≤ 2 and adequate hematologic, renal and liver
function. Patients will receive 1x107 pfu HSV1716 through their
pleural catheter on one, two or four occasions a week apart,
in three separate patient cohorts. An extension cohort of three
iMig2016.ORG
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ABSTRACT BOOK
patients will be treated at the highest tolerated dose. The primary objectives are to determine the safety and tolerability of
HSV1716 given intrapleurally in patients with inoperable MPM.
Detailed safety analyses will be undertaken. The secondary objective is to obtain evidence of HSV1716 replication/persistence
and patient immune responses through analysis of pleural fluid
collected on alternate days for one week, after the last HSV1716
administration, then weekly. An exploratory objective will be to
assess tumour response by CT using modified RECIST criteria.
Results: Three patients have received a single dose of HSV1716
through their IPC, three have received two doses and four
have received four doses with recruitment of an additional two
patients required at the four dose level to complete the trial.
HSV1716 is well-tolerated with a limited number of possibly
related adverse events identified. There is evidence of HSV1716
replication/persistence in most patients and the potential for
anti-tumour immune responses has also been observed in
patients.
Conclusion: Two patients receiving 4 doses of HSV1716 are
required to complete the study and a randomised phase II study
of intrapleural HSV1716 is under consideration.
Keywords: intrapleural, Immunotherapy, oncolytic HSV, clinical study
20-85 years with chemotherapy-resistant malignant pleural
mesothelioma who had not received chemotherapy in the last
6 weeks, had an Eastern Cooperative Oncology Group performance status of 0-1, had certain functions of bone marrow,
liver, kidney, and lung at the screening visit, had evaluable or
measurable lesion with CT or MRI, had lesion which can be
administrated with HVJ-E, and a life expectancy of >8 weeks.
Exclusion criteria were presence of central nervous system
metastases, autoimmune disease, interstitial pneumonia or pulmonary fibrosis needed to treatment, hemostatic disorder, and
other malignant lesions, use of systemic glucocorticosteroids,
a history of treatment with other investigational products last 4
weeks before the informed consent. The protocol is consistent
of initial intra-tumoral administration of HVJ-E and the subsequent three subcutaneous administration within two weeks and
then washed out for two weeks. This one cycle will be repeated
twice. HVJ-E will be given at a lower dose for each injection. If
dose-escalation is permitted by independent data monitoring
committee, another cohort will be given at a higher dose of
each injection.
Results: This phase I clinical trial is now in progress.
Conclusion: We will make a presentation of the detail about
this trial, and the preclinical study of HVJ-E.
Keywords: immunotherapy, HVJ, HVJ-derived nanoparticle
MS11: CRITICAL SIGNALING PATHWAY
14:15 – 15:45
MS10.07: A PHASE I CLINICAL TRIAL OF HVJDERIVED NANOPARTICLE FOR CHEMOTHERAPYRESISTANT MALIGNANT PLEURAL
MESOTHELIOMA
Chunman Lee1, Atsuhiro Saito1, Yoshihisa Kadota2, Shinji Atagi3 , Takashi Nakano4 , Yasufumi Kaneda5, Meinoshin Okumura6
Medical Ctr. For Translational Research, Osaka University,
SUITA, JAPAN, 2Surgery, Osaka Prefectual Medical Center for
respiratoy and allergic diseases, Habikino, JAPAN, 3Kinki-Chuo
Chest Medical Center, Sakai, JAPAN, 4Hyogo College of Medicine,
Nishinomiya, JAPAN, 5Division of Gene Therapy Science, Osaka
University, Suita, JAPAN, 6Department of Thoracic Surgery, Osaka
University, Suita, JAPAN
1
Objectives: Hemagglutinating Virus of Japan Envelope
(HVJ-E): HVJ-derived nanoparticle, possess the various antitumor activities whose mechanism is different from chemotherapy. One is enhancing multiple antitumor immunities such
as activation of dendritic cells, induction of natural killer cells
and CTL, and suppression of regulatory T cells. Other activities
are direct tumor-killing by the induction of cell death through
the RIG-I/MAVS pathway. We examined the actual antitumor
activities of HVJ-E on the orthotopic implantation model, and
HVJ-E had significant antitumor activity compared with control
group. We therefore do the phase I dose escalation safety/
tolerability and preliminary efficacy study of intra-tumoral and
subcutaneous administration of HVJ-E in patients suffering
from chemotherapy-resistant malignant pleural mesothelioma
for clinical applications.
Methods: In this phase I trial, we are recruiting patients aged
MS11.01: ANTAGONIZING THE HEDGEHOG
PATHWAY WITH VISMODEGIB IMPAIRS
MALIGNANT PLEURAL MESOTHELIOMA GROWTH
IN VIVO BY AFFECTING STROMA
Mayura Meerang1, Karima Bérard1, Emanuela Felley-Bosco2,
Olivia Lauk1, Bart Vrugt3 , Andreas Boss4 , David Kenkel4 , Angela
Broggini-Tenzer5, Rolf Stahel6 , Stephan Arni1, Walter Weder1,
Isabelle Opitz1
SWITZERLAND, 2Department of Molecular Oncology, University
Hospital Zurich, Zurich, SWITZERLAND, 3Institute of Surgical
Pathology, University Hospital Zurich, Zurich, SWITZERLAND, 4Institute of Diagnostic And Interventional Radiology, University
Hospital Zurich, Zurich, SWITZERLAND, 5Laboratory For Molecular Radiobiology, Radiation Oncology, University Hospital Zurich,
Zurich, SWITZERLAND, 6Clinic For Oncology, University Hospital
Zurich, Zurich, SWITZERLAND
1
Objectives: Upregulation of the Hedgehog (Hh) signaling
pathway is associated with poor prognosis of malignant pleural
mesothelioma (MPM) patients. An autocrine driven upregulation of the Hh pathway was described in MPM, in which the
ligand, desert hedgehog (DHH), was produced from tumor cells.
Paracrine activation of Hh signaling has been described in other
iMig2016.ORG
50
ABSTRACT BOOK
solid tumors and our recent investigation revealed that the Hh
pathway was activated in both tumor and stroma of human
MPM specimens. In this study, we investigated the importance
of paracrine Hh signaling in MPM progression in vivo.
Methods: We employed an orthotopic immunocompetent
rat MPM model. Sarcomatoid rat MPM cells transfected with
luciferase (IL45-luc) were implanted subpleurally in Fischer
rats. After tumors were formed, rats were treated orally with a
FDA approved Hh antagonist, vismodegib, (once daily, 100 mg/
kg; n=6) for 6 days while control group (n=6) received vehicle
alone. Tumor load was monitored by bioluminescence, magnetic resonance imaging (MRI) and macroscopically. Tumors were
harvested and evaluated for Hh target genes expression by
quantitative real time PCR and immunohistochemistry. Tumor
cells were isolated and cultured in medium without serum at
37°C, 5%CO2 and 3%O2. Tumor cell culture supernatant was
collected freshly and applied to confluent mouse embryonic fibroblasts (NIH3T3). The expression of Hh pathway target genes
in NIH3T3 cells were analysed at 72h afterwards by quantitative
real time PCR.
Results: Similar to that observed in human MPM specimens,
positive immunohistochemical staining of Hh pathway components, Glioma associated oncogene 1 (GLI1) and Patched1
(PTCH1), were detected in both tumor and stromal fractions
of the rat MPM model. Hh ligand, DHH, was predominantly
expressed in the tumor fractions. Daily treatment with vismodegib in vivo efficiently downregulated Hh target genes, Gli1,
Hedgehog Interacting Protein (Hhip) and Ptch1, and caused a
significant reduction of tumor volume, and tumor growth delay.
Tumor cell proliferation, Ki-67 and phospho-histoneH3 positive
indices, were significantly reduced in the treated group. Immunohistochemical analyses revealed that vismodegib treatment
primarily down regulated Hh target genes, GLI1 and HHIP, in
the stromal compartment along with a reduced expression of
previously described fibroblast Hh responsive genes such as
Fibronectin (Fn1) and vascular endothelial growth factor (Vegf ).
Primary cells isolated from the rat tumors cultured in physiological O2 level (3%) continued to express Dhh but did not respond
to vismodegib treatment in vitro. However, culture supernatant
from these cells stimulated Gli1, Ptch1, and Fn1 expression in
mouse fibroblasts NIH3T3 which was suppressed by vismodegib treatment.
Conclusion: MPM cells expressed ligand and induced Hh
response in fibroblasts, implying the role of paracrine Hh
signaling in MPM. Hh pathway activated fibroblasts in turn
produced growth factors important for tumor progression. Hh
pathway antagonization in MPM stroma efficiently delayed
tumor cell growth, emphasizing the importance of Hh pathway
as a treatment target for MPM. In addition, our study highlights
the significant aspect of tumor-stroma crosstalk in promoting
MPM progression.
Keywords: Paracrine, Orthotopic rat mesothelioma model,
Hedgehog signaling pathway, Stroma
MS11.02: SIRT1 AT THE CROSSROADS OF
AKT1 AND ERβ IN MALIGNANT PLEURAL
MESOTHELIOMA CELLS
Giulia Pinton1, Sara Zonca1, Arcangela G. Manente1, Maria
Cavaletto2, Ester Borroni3 , Antonio Daga4 , Puthen V. Jithesh5,
Dean Fennell6 , Stefan Nilsson7, Laura Moro1
Novara, ITALY, 2Sciences And Technological Innovation, University of Piemonte Orientale, Alessandria, ITALY, 3Health Sciences,
University of Piemonte Orientale, Novara, ITALY, 4IRCCS San
Martino-IST, Genova, ITALY, 5Sidra Medical and Research Center,
Doha, QATAR, 6Cancer Studies, University of Leicester, Leicester,
UNITED KINGDOM, 7Biosciences And Nutrition, Karolinska Institutet, Huddinge, SWEDEN
1
Objectives: Characterize the role of AKT isoforms in human
malignant pleural mesothelioma (MPM).
Methods: We have evaluated the expression and function of
AKT isoforms in MPM tumor samples and MPM derived cell
lines.
Results: Here we show that MPM patients whose tumors
express high levels of AKT1 exhibit a significantly worse prognosis, whereas no significant correlation with AKT3 expression
is observed. We provide data that establish a phosphorylation
independent role of AKT1 in affecting MPM cell shape and anchorage independent cell growth in vitro and highlight the AKT1
isoform-specific nature of these effects. We describe that AKT1
activity is inhibited by the loss of SIRT1-mediated deacetylation
and identify, by mass spectrometry, 11 unique proteins that
interact with acetylated AKT1. Our data demonstrate a role of
the AKT1/SIRT1/FOXM1 axis in the expression of the tumor
suppressor ERβ. We further demonstrate an inhibitory feedback
loop by ERβ, activated by the selective agonist KB9520, on this
axis both in vitro and in vivo.
Conclusion: Our data broaden the current knowledge of ERβ
and AKT isoform-specific functions that could be valuable in the
design of novel and effective therapeutic strategies for MPM.
Keywords: Malignant pleural mesothelioma, AKT isoforms,
ERbeta
MS11.03: EVALUATION OF SENSITIVITY TO PI3K/
MTOR AND FAK INHIBITION IN PRE-CLINICAL
MODELS OF MALIGNANT MESOTHELIOMA
Ian R. Powley1, Xiao-Ming Sun1, Tatyana Chernova1, Sara
Galavotti1, Stefano Grosso1, Joaquin Zacarias-Cabeza1, John Le
Quesne2, Jonathan Bennett2, Apostolos Nakas2, J. H. Pringle3 ,
Anne E. Willis1, Dean Fennell2, Marion Macfarlane1
UNITED KINGDOM, 3University of Leicester, Leicester, UNITED
KINGDOM
1
iMig2016.ORG
51
ABSTRACT BOOK
Objectives: Despite the increasing prevalence of Malignant
Mesothelioma (MM), there remains a paucity of approved,
effective therapy for this cancer. There is therefore an urgent
need for novel treatments targeting this disease. Recent studies
of MM cell lines and primary tumours revealed a high level
constitutive activation of both the phosphotidylinoside 3 kinase
(PI3K) and Focal Adhesion Kinase (FAK) signalling pathways;
events which have previously been associated with poor patient
prognosis. Therefore, dual inhibition of these key pathways may
prove beneficial in targeting MM cells for growth arrest and/or
apoptosis. The objective of this study is to evaluate the sensitivity of pre-clinical MM patient models to PI3K/mTOR and FAK
inhibition, and correlate this to genetic and other biomarkers.
Methods: Patient-derived MM cell lines were established
(Chernova, Sun et al, Cell Death Differ., in press) and used to
evaluate the efficacy of the pan-PI3K/mTOR inhibitor VS-5584
and the FAK inhibitor VS-6063, which are both in clinical trials
for MM, in 2D and 3D culture models. Cell cycle status and
apoptotic cell death were assayed by flow cytometry using
propidium iodide incorporation and phosphatidylserine externalisation and/or Caspase-Glo® assay, respectively. Inhibition
of cell signalling pathways was assessed by western blotting. In
addition, using an ex-vivo 3D tumour explant model that retains
the tumour microenvironment, we have treated freshly resected MM patient explants with VS-5584 and/or VS-6063 and
assessed on-target drug effects and induction of apoptosis by
histology and immunohistochemistry.
Results: In 2D culture, treatment of patient-derived MM cell
lines with VS-5584 and VS-6063 results in a rapid inhibition of
their respective signalling pathways and concomitant inhibition
of cell proliferation and induction of cell cycle arrest, without induction of apoptosis. In contrast, combination treatment of both
3D tumour explants ex vivo and cells cultured in a 3D matrigel
model with VS-6063, results in inhibition of FAK signalling and
corresponding induction of apoptosis.
Conclusion: In a MM patient-derived ex-vivo 3D explant model,
VS-6063 both as a single agent and in combination with VS5584 induces apoptotic cell death; this platform enables us
to correlate drug sensitivity to genetic and other biomarkers
and has the potential to permit molecular stratification of MM
patients for future clinical trials.
Keywords: FAK, PI3K/mTOR, Cell Death, Pre-Clinical Models
are restored using mimics. Results from our lab have led to the
world’s first clinical trial of a microRNA replacement strategy in
thoracic cancer patients, currently nearing the end of Phase I.
The study presented here consisted of a head-to-head comparison of microRNA mimics and aimed at identifying the most potent microRNAs for future development as therapeutic agents.
Methods: Synthetic mimics were designed based on mirbase
sequences of previously reported tumour suppressor microRNAs: miR-1, miR-15a, miR-15b, miR-16, miR-29c*, miR-31,
miR-34a, miR-34b, miR-34c, miR-126, miR-137, miR-145 and
miR-193a-3p. These were used at three concentrations (1.1, 3.3
and 10 nM) to transfect a panel of MPM cell lines (MSTO-H211,
H2052, H28, MM05, VMC23), and effects on growth were measured using standard proliferation and colony formation assays.
The most growth inhibitory microRNA mimics were further investigated to understand mechanism of growth inhibition using
apoptosis, cell cycle, senescence and migration assays. Active
mimics were used together to identify synergistic combinations,
with bioinformatics used to predict pathways affected by mimic
combinations.
Results: Of the microRNAs previously reported to have growth
inhibitory activity, mimics corresponding to miR-15a, miR-15b,
miR-16, miR-34a, miR-34b, miR-34c, miR-137 and miR-193a-3p
were strongly growth inhibitory in all cell lines, showing greater
than 50 % growth inhibition at a concentration of 3.3 nM,
with miR-137 the most active overall in all cell lines tested. In
contrast, miR-126 was highly active in only 2 of the investigated
cell lines, whereas the other microRNA mimics were modestly
active at 10 nM or inactive at all concentrations used. Induction of apoptosis and cell cycle arrest were the most common
mechanisms of action. The highly active microRNAs had multiple common target genes involved in cell cycle and apoptosis,
and these targets were downregulated following transfection
of the mimics. Synergistic activity of mimic combinations was
exemplified by the combination of miR-16 with miR-193a-3p,
which exceeded the activity of either mimic alone. In general,
microRNAs with less overlap in target genes yielded the greatest combinatorial effect.
Conclusion: Multiple microRNAs exhibit tumour suppressor
characteristics with growth inhibitory activity, and this head-tohead comparison reveals miR-137 to be the most effective at
inhibiting MPM cell growth in vitro. This, and combinations such
as miR-16 with miR-193a-3p, represent candidates for future
clinical development as therapeutic agents.
Keywords: tumour suppressor, microRNA
MS11.04: IDENTIFYING MICRORNAS WITH
THERAPEUTIC POTENTIAL IN MALIGNANT
Glen Reid, Andrew Della Gatta, Hyerim Suh, Marissa Williams,
Yuen Yee Cheng, Ruby Lin, Nico Van Zandwijk
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA
Objectives: MicroRNA expression is globally downregulated
in cancers including malignant pleural mesothelioma (MPM).
We and others have shown that multiple microRNAs have
tumour suppressor activity in MPM cell lines when the levels
MS11.05: MICRORNA-31 REGULATES
CHEMOSENSITIVITY IN MALIGNANT PLEURAL
MESOTHELIOMA VIA ALTERED INTRACELLULAR
DRUG LOCALISATION
Hannah L. Moody1, Michael J. Lind2, Stephen G. Maher3
School of Biological, Biomedical And Environmental Sciences, University of Hull, Hull York Medical School, Hull, UNITED
KINGDOM, 2Centre For Oncology And Haematology, Hull and
1
iMig2016.ORG
52
ABSTRACT BOOK
East Yorkshire NHS Trust, Hull, UNITED KINGDOM, 3School of
Biological, Biomedical And Environmental Sciences, University of
Hull, University of Hull, Hull, UNITED KINGDOM
Objectives: Malignant pleural mesothelioma (MPM) is associated with extremely poor prognosis and many patients are
unresponsive to treatment; developing resistance to chemotherapeutics. MicroRNA (miRNA/miR) are small, non-coding
RNA that function to regulate gene expression and have been
demonstrated to alter cellular sensitivity to cytotoxic agents in
various cancers. MiR-31 is encoded on chromosome 9p21.3,
which is reportedly the most frequently deleted genomic location in MPM tumours. Here, we examined if dysregulation of
miR-31 alters MPM chemosensitivity.
Methods: The miR-31 deficient NCI-H2452 and miR-31 positive
P31 epithelioid cell lines were used as in vitro models of MPM.
Stable miR-31 overexpression or suppression was achieved via
liposomal-based transfection of plasmid-based vectors and antibiotic selection. Clonogenic assay was employed as a measure
of cellular sensitivity to cisplatin and carboplatin; colonies were
enumerated in an unbiased manner using a GelCount device.
Inductively coupled plasma mass spectrometry (ICP-MS) was
utilised to quantify cellular cisplatin flux. Subcellular fractionation via sucrose gradient centrifugation was used to isolate
intracellular organelles. Immunofluorescent microscopy was
used to visualise intracellular localisation and protein density.
Gene expression was assessed by qPCR and protein expression
by Western Blot.
Results: Surprisingly, reintroduction of miR-31 into cisplatinand carboplatin-treated NCI-H2452 cells significantly increased
chemoresistance compared to vector controls; conversely,
suppression of miR-31 in P31 cells increased cellular sensitivity
to cisplatin. Additionally, miR-31 reintroduction mediated a
delay in the cytotoxic activity of chemotherapy. Interestingly,
a higher relative intracellular concentration of platinum was
observed in miR-31 transfected cells, potentially as a result of
increased expression of the plasma membrane-bound cisplatin
influx transporter CTR1. However, a concurrently significantly
decreased intranuclear concentration of platinum was determined in miR-31 expressing cells, suggesting altered nuclear
transport or sequestration within the cytosolic compartment.
Consequently, DNA damage was found to be greatly reduced
in miR-31 expressing NCI-H2452 cells, and antagonistically
higher in miR-31 suppressed P31 cells, supporting the alteration in transit to the nucleus. Subsequently, we identify that
a miR-31-mediated increase in the lysosomal-associated drug
transporter ABCB9, in conjunction with reduced expression
of the associated bipotential transcription factor OCT1, may
promote the extranuclear sequestration of chemotherapeutic
agents in lysosomes in MPM cells expressing miR-31.
Conclusion: Here, miR-31 expression was found to significantly enhance chemoresistance in MPM cells in vitro. While
deletions in the genomic location encoding miR-31, 9p21.3, may
be associated with an overall poor prognosis, the loss of miR-31
may not actually contribute to the chemoresistance observed in
MPM patients. Our current work further examines the impact of
how miR-31 functionally modulates intracellular spatial distribution of cisplatin. Additionally, the manipulation of both OCT1
and ABCB9 expression is ongoing in order to determine the
relative contributions of these molecules to the chemoresistant
phenotype observed.
Keywords: microRNA, ABCB9, DNA damage, chemotherapy
MS11.06: TARGETING THE RATE-LIMITING
STEP OF PROTEIN SYNTHESIS OVERCOMES
CHEMORESISTANCE IN MALIGNANT
MESOTHELIOMA
Stefano Grosso1, Kate Dudek1, Carolyn Jones1, Jack Godfrey1,
Ruth Spriggs1, Ania Wilczynska1, Tatyana Chernova1, Xiao-Ming
Sun1, Gareth J. Miles1, David Dinsdale1, Jonathan Bennett2,
Apostolos Nakas2, John Le Quesne1, Kelvin Cain1, Marion Macfarlane1, Martin Bushell1, Anne E. Willis1
UNITED KINGDOM
1
Objectives: Translation of mRNA into protein is a metabolic energy demanding process. As a consequence, protein synthesis
is highly regulated, mainly at the stage of initiation; a process
that requires eukaryotic initiation factors (eIFs) to recruit the
mRNA to the ribosomes. eIFs are downstream targets of signal
transduction pathways activated by growth factor stimulation,
first leading to an increase of translation and cell size, then to
cell division, finally to tumour formation. The objective of this
study is to modulate and re-shape tumour cell protein synthesis
to prevent cancer progression.
Methods: Malignant Mesothelioma (MM) primary cell lines
were established from freshly resected patient tumours
(Chernova, Sun et al., Cell Death Differ., in press). Transcription,
translation and miRNA expression were analysed by specific
arrays, followed by bioinformatics analysis. To analyse translation status ribosomes from MM primary cell lines (in addition to
untransformed controls) were separated on sucrose gradients
and translating mRNAs were analysed with an unbiased array
approach. Northern and western blot were used to confirm the
proteins overexpressed in MM patients compared to healthy
control mesothelial cells. Candidate novel biomarkers were
identified and their biological role in MM was evaluated.
Results: Analysis of the transcriptome and the translatome of
MM primary cells relative to untransformed control identified
distinct changes in tumors samples in subset of mRNAs (Fig
1A). Our data showed that the upregulation of translation (Fig
1B) was, in part, driven by increased expression of factors
required for both the initiation and elongation stages of proteins
synthesis (Grosso et al., PLoS One, 2011; Miluzio et al., Oncotarget, 2015). Interestingly, there was a direct correlation between
eIFs expression and the degree of MM sensitivity to cisplatin
treatment. Furthermore, the data suggest an increase in polyribosomal association of mRNAs involved in the mitochondrial
stress response and a corresponding increase in synthesis of
these proteins in MM. Finally, bioinformatics analysis allowed
the identification of 5’UTR consensus motifs in the mRNAs that
were preferentially translated in MM cells. iMig2016.ORG
53
ABSTRACT BOOK
Conclusion: Alteration of signalling pathways upstream of the
translational machinery and/or the expression of key translation
initiation factors modulates the response of MM tumour cells
to cisplatin-based chemotherapy. The link between translation
and the energetic metabolism of this tumour is currently under
consideration. Taken together, these studies may elucidate new
strategies to treat this disease.
Keywords: miRNA, Translation, Transcription, Ribosome
MS11.07: THE ROLE OF MONOCYTE CHEMOTACTIC
PROTEIN-1 (MCP-1) IN MESOTHELIOMA-INDUCED
MALIGNANT PLEURAL EFFUSION FORMATION
Sally M. Lansley1, Hui Min Cheah2, Catherine A. Rinaldi3 , Jenette Creaney3 , Yc Gary Lee4
Pleural Medicine Unit, Institute for Respiratory Health, Perth,
WA, AUSTRALIA, 2Institute for Respiratory Health and School
of Medicine & Pharmacology, University of Western Australia,
Perth, WA, AUSTRALIA, 3School of Medicine and Pharmacology,
National Centre Of Asbestos Related Diseases, University of
Western Australia, Nedlands, WA, AUSTRALIA, 4Institute for Respiratory Health, School of Medicine & Pharmacology, University
of Western Australia and Department of Respiratory Medicine, Sir
Charles Gairdner Hospital, Perth, WA, AUSTRALIA
1
Objectives: More than 90% of malignant pleural mesothelioma (MPM) patients present with a pleural effusion. Current
methods of managing effusions are limited. Therefore, identifying mechanisms of malignant effusion formation may provide
novel therapeutic options. We recently demonstrated that MCP1 plays a major role in the formation of fibrinolytic-induced
pleural effusion. Others have established MCP-1 as a driver of
malignant pleural effusion in a mouse model of lung cancer.
We therefore aimed to determine whether MCP-1 contributes to
MPM effusion development.
Methods: Expression of MCP-1 mRNA and protein in human
(n=11) and mouse (n=9) mesothelioma cell lines was measured
by RT-PCR and MCP-1 protein expression and distribution in
human and mouse cells examined by immunocytochemistry.
MCP-1 levels were quantified by ELISA in pleural fluid supernatants from 197 patients. Pleural fluid samples (n=298) collected longitudinally from MPM patients were also assessed for
iMig2016.ORG
54
ABSTRACT BOOK
changes in MCP-1 levels. In vivo effects of MCP-1 inhibition on
MPM-induced pleural effusion were examined using an MCP-1
receptor (CCR2) antagonist (up to 6 daily injections) or MCP-1
neutralising antibody (4 daily injections) delivered intraperitoneally.
Results: All mouse and human mesothelioma cell lines tested
expressed MCP-1 mRNA and protein. MCP-1 protein was
distributed within the nucleus and cytoplasm of mesothelioma
cells. MCP-1 levels were significantly higher in MPM pleural
effusions (n=78) than in non-MPM pleural fluids (n=119; from
metastatic pleural carcinomas/benign causes): median 1140 vs
450 pg/ml respectively, p =0.02. Longitudinal measurements of
pleural fluid MCP-1 levels in 35 patients showed a significant
increase in expression (0.37±0.13 pg/ml/100 days, p =0.005)
as tumour progressed. Extensive pleural mesothelioma tumour
and large pleural effusions were induced in CBA mice (n=63)
by intrapleural injection of murine mesothelioma (AC29) cells.
CCR2 antagonist treatment significantly reduced pleural
effusion volumes (median (IQR); controls: 843 (583-1234)
μL vs treatment group 270 (146-835) μL, p =0.03. Treatment
with MCP-1 neutralising antibody also significantly reduced
pleural effusion volumes (mean±SEM; 72.9±12.4 μL) compared
to IgG isotype (174.0±39.8 μL) and saline (127.4±10.3 μL)
controls, p =0.03. Tumour weights did not differ significantly
between any treatment groups.
Conclusion: MCP-1 is significantly over-expressed in MPM
effusions and inhibition of MCP-1 activity potently reduced the
formation of MPM effusion. MCP-1 represents a potential therapeutic target to control MPM malignant effusions.
Keywords: MCP-1, Pleural effusion
MS12: TREATMENT ADVANCES IN PERITONEAL
MESOTHELIOMA / PALLIATIVE CARE
FOR ALL MESOTHELIOMA
14:15 – 15:45
MS12.01: PERITONEAL MESOTHELIOMA: PHASE-II
TRIAL OF TAILORED SYSTEMIC CHEMOTHERAPY
BASED ON SENSITIVITY TESTS ON PRIMARY CELL
CULTURES
Dario Baratti1, Shigeki Kusamura1, Rossella Bertulli2, Federica Perrone3 , Antonello D. Cabras3 , Marzia Pennati4 , Marcello
Guaglio1, Nadia Zaffaroni4 , Marcello Deraco1
Surgery, Fondazione IRCCS Istituto Nazionale Tumori, Milano,
ITALY, 2Medical Oncology, Fondazione IRCCS Istituto Nazionale
Tumori, Milano, ITALY, 3Pathology, Fondazione IRCCS Istituto
Nazionale Tumori, Milano, ITALY, 4Experimental Oncology, Fondazione IRCCS Istituto Nazionale Tumori, Milano, ITALY
1
tive surgery and hyperthermic intra-peritoneal chemotherapy
(CRS/HIPEC). However, the comprehensive management of this
disease has yet to be optimized, and the role of perioperative
systemic chemotherapy (s-CT) is still poorly defined. The aim
of this phase-II trial was to assess the impact of individualized
postoperative s-CT in DMPM patients treated with CRS/HIPEC,
based on the chemosensitivity profile on primary cell cultures.
Methods: CRS involved peritonectomy procedure and mutivisceral resections, according to a standardized technique. HIPEC
was performed with cisplatin plus doxorubicin. In each patient,
primary cell cultures were obtained from DMPM surgical specimens. Chemosensitivity was determined in vitro by a proliferative assay, based on 3H-thymidine incorporation. Cytotoxic
(cisplatin, carboplatin, pemetrexed, gemcitabine, vinorelbine,
doxorubicin, vincristine), and molecularly targeted agents (everolimus, sorafenib) were tested. Cell proliferation was assessed
by immunohistochemical staining of Ki-67 nuclear antigen with
monoclonal antibody MIB-1. s-CT was planned within 8 weeks
after CRS/HIPEC. Overall survival was the primary study endpoint.
Results: From January 2012 to October 2015, 38 consecutive
s-CT naïve patients were enrolled in the study. CRS/HIPEC was
performed in 33 patients. The quality of surgical cytoreduction
was rated as adequate (residual disease nodules ≤ 2.5 mm) in
28 of them, and grossly incomplete in 5. Only palliative/debulking surgery (and no HIPEC) was possible in five. Operative death
occurred in one patient and severe complications in 15 (39.5%).
At pathological examination, epithelial DMPM was diagnosed
in 33 patients, and biphasic DMPM in 5; positive lymph-nodes
were found in 5 patients. Primary cell cultures could not be
obtained in 13 patients, due to poor cell vitality. Cultures
were not assessable for chemosensitivity in 15, due to insufficient 3H-thymidine incorporation. Of the remaining 10 patients,
six were resistant to all tested drugs, and four were sensitive to
≥1 drug (everolimus, doxorubicin, gemcitabine, cisplatin, vincristine). Proliferative activity was relatively low, with a median
percentage of Ki-67-expressing cells of 18.2% (range 5-45%).
Thirteen of 28 patients undergoing adequate CRS/HIPEC did
not receive postoperative s-CT, due to operative death (n=1),
or poor condition/refusal (n=12). Fiveteen were treated with
cisplatin/carboplatin and pemetrexed. Chemosensitivity tests
were available and multiresistant in 4 of them Median follow-up
was 10.7 months (range 1-34.4). Two-year survival was 46.4%
in the overall series, and 62.6% in those undergoing adequate
CRS/HIPEC. The completeness of cytoreduction was the only
significant prognostic predictor (P=0.003, log-rank).
Conclusion: Our findings confirm the poor sensitivity of DMPM
to systemic agents and do not support the role of currently
available s-CT. The malignant potential of the disease appears
to be related to its chemoresistance, rather than elevated
cellular proliferation. Prognostic improvements may depend on
aggressive comprehensive local-regional management. Better
understanding of DMPM molecular features, and development
of new systemic and/or targeted therapies are needed.
Keywords: systemic chemotherapy, Peritoneal mesothelioma,
targeted therapy, HIPEC
Objectives: The prognosis of diffuse malignant peritoneal
mesothelioma (DMPM) has recently improved with cytoreduciMig2016.ORG
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ABSTRACT BOOK
MS12.02: GENDER AND MENOPAUSAL STATUS:
SURVIVAL IN PERITONEAL MESOTHELIOMA
women. With increased survival for premenopausal women, it
would be important to better understand the role of ERβ and
Ki-67 in mesothelioma.
Keywords: Survival, Estrogen, ERβ, Gender
Yaakov Bressler, Gleneara E. Bates, Robert N. Taub
Medicine, Columbia University Medical Center, New York, NY,
Objectives: It is well recognized though not fully understood
that women live longer than men with mesothelioma. We have
investigated survival difference related to gender and pre- and
post-menopausal status in peritoneal mesothelioma. We wished
to determine whether there are survival differences related to
gender, age, and menopause.
Methods: An IRB approved retrospective analysis of mesothelioma patients treated from 1990 – 2015 was done at CUMC.
Patients with peritoneal diagnoses and recipients of CRS and
HIPEC were included. Patient characteristics (gender, histology, and pre/post menopausal status) were collected. Survival
and prognosis analyses were performed using Kaplan-Meier
curves and univariate cox proportional hazards model. Other
established prognostic factors were evaluated with multivariate
analysis.
Results: Median survival time of all MPM patients (n=195) was
3.21 years with (95% CI: 2.38- 5.53), with median follow-up of
3.44 years (SD=3.4, minimum=0.014 and maximum=16.752)
years from first operation. Patient set included 111 men
(57%) and 84 women (43.1%) with female sex having favorable survival [HR: 0.442 95% (CI: 0.296-0.659), p<0.001] of
110.1 months with (95% CI: lower bond: 48.3). Mean age at
diagnosis was 54.8 years [HR: 1.027 (95% CI: 1.012-1.042)]
with 111 men (57%) and 84 women (43.1), with female gender
having favorable survival [HR: 0.442 95% CI: 0.296-0.659)].
Majority of patients had epithelioid histology (n=161(82.6%),
with the remainder biphasic/sarcomatoid (n=34, 17.4%), with
increased risk of death with non-epithelioid histology [HR:
2.46(95%CI 1.59-3.82)], (P=0.001). Median age for all female
MPM patients (n=84) was 53 years (SD=14.5), minimum=14.7,
maximum=79.9. Median survival was 110.1 months (95% confidence interval lower bound: 48.3) months, mean survival=93.4
(SE=8.69). Median Age of premenopausal patients was 36 years
(SD=9.26), minimum=14.7, maximum=48.1. Median survival
cannot be determined as at follow up of 110 months, survival
was 72% (n=23).
Conclusion: These data show that women with MPM have a
better survival than men and that premenopausal women have
better survival than either men or postmenopausal women. No
significant genetic difference between gender in MPM has been
identified, aside from an increased TP53 mutation – a result not
yet fully understood. Estrogen receptor β (n) has shown to be
expressed (with IHC >6) in 15% of mesotheliomas and is associated with an increased survival in MPM. Receptor frequency
between genders has demonstrated the level of ERβ in women
to be triple that of men, with higher expression in post-menopausal women than pre-menopausal women. Activation of ERβ
from estradiol may contribute to survival differences. Data has
shown that administration of KB9520, an ERβ agonist, leads to
less aggressive tumors and sensitization of tumors to chemotherapy. Further, Ki-67, a biomarker for cellular proliferative
activity where low Ki-67 is associated with increased survival,
has been found to be expressed in men twice as much as in
MS12.03: SIMULTANEOUS CARE (SIMC) IN
MESOTHELIOMA (MM): A DEDICATED TEAM TO
PREVENT URGENT AND UNPLANNED HOSPITAL
ADMISSIONS
Giulia Gallizzi1, Federica Grosso1, Alma Kasa2, Barbara Oneglia2, Giacomo Taverna3 , Fausto Pernazza4 , Paola Ballarino2,
Annalisa Roveta5, Liana Todisco6 , Silvia Zai1, Ezio Piccolini7, Alberto Muzio8 , Gianmauro Numico1, Massimo D’Angelo9, Daniela
Degiovanni10
Oncology Unit, SS. Antonio e Biagio e C. Arrigo, Hospital,
Alessandria, ITALY, 2Hospice Monsignor Zaccheo / Uocp, Santo
Spirito, Hospital, Casale Monferrato, ITALY, 3Radiology Unit,
Santo Spirito, Hospital, Casale Monferrato, ITALY, 4Chirurgia
Toracica, SS. Antonio e Biagio e C. Arrigo, Hospital, Alessandria,
ITALY, 5Ssa Sviluppo E Promozione Scientifica, SS. Antonio e
Biagio e C. Arrigo, Hospital, Alessandria, ITALY, 6Radiotherapy
Unit, SS. Antonio e Biagio e C. Arrigo, Hospital, Alessandria,
ITALY, 7Pneumologia, Santo Spirito, Hospital, Casale Monferrato,
ITALY, 8Oncology Unit, Santo Spirito, Hospital, Casale Monferrato, ITALY, 9Centro Sanitario Amianto, Azienda Sanitaria Locale
AL, Casale Monferrato, ITALY, 10 Ss Hospice Monsignor Zaccheo/
Uocp, Santo Spirito, Hospital, Casale Monferrato, ITALY
1
Objectives: MM is a fatal cancer with great symptom burden
and treatment has only a modest impact on survival. Palliative
care is crucial in clinical management and helps patients and
families dealing with disease related symptoms and psychological implications. In 2012 the Italian Centro Controllo per le
Malattie (CCM) supported a project that aimed at assuring Early
Palliative Care (SimC) and psychological support to newly diagnosed patients and families in order to improve quality of care
and reduce unplanned Accident and Emergency Department
(A&E) admissions. Here we report on the preliminary results
of the SimC Program at a referral centre in the high asbestos
polluted area of Casale Monferrato.
Methods: An Integrated Palliative Care Team, available 8 am
to 10 pm, 7 days a week, followed each patient since diagnosis
throughout the whole course of the disease, both at home and
in every other place. The most relevant clinical variables and
symptoms were registered into a dedicated web database.
Specific questionnaires were delivered to collect data about
awareness of the disease and satisfaction regarding the care
after the cessation of SimC.
Results: Since 4/2013 to 10/2015, 79 MPM patients, 39 M
(49%) and 40 F (51%), median age 71 (IQR 65-78; range 53-99)
were included. PS according to Karnofsky was 30-40 in 38
(48%), 50-60 in 38 (48%) and >70 in 2 (4%) pts. The median
duration of the SimC support was 33 days (mean 56; range
10-465). At the time of inclusion 39 patients (49%) complained
with pain, median NRS 4.5 (range 2-9). Other reported sympiMig2016.ORG
56
ABSTRACT BOOK
toms were dyspnea in 58 (73%), fatigue in 50 (63%), cough in
24 (30%), peripheral oedema in 24 (30%), agitation/confusion
in 17 (22%) patients. Sixty-three (79%) patients were perfectly
aware about their disease and 13 (17%) about their terminal
condition. Seventy-four families (94%) were fully informed
about the disease and 71 (90%) about the imminent fatal
outcome. SimC support ended due to the following causes:
death at home for 53 patients (67%), improvement of symptoms in 9 patients (10%), admission to the palliative care ward
in 13 patients (16%). Five patients (6%) were hospitalized for
planned palliative procedures. Only one patient (1.2%) required
A&E admission due to high fever from pneumonia. The questionnaire highlighted that 96% of patients had no pain in the
last 24 hrs of life, 94% received analgesic treatment, and none
of the patients underwent resuscitation manoeuvres. Death was
not an unexpected event in 98%. The palliative care team had
a contact with the family in the last 24 hours and was informed
about death in 96% of patients.
Conclusion: This study focused on SimC in MM. Data is still
partial and we are working to retrieve it. The most relevant
achievements are that the vast majority of patients (98.8%)
followed within this program did not require any urgent admission to A&E Department and died at home according to their
preference, with careful and assiduous support by the palliative
care team, in an appropriate context of Public Health Service.
Keywords: Psycological support, palliative care, Advanced
mesothelioma, Simultaneous care
MS13: GENOMICS AND DRUG SENSITIVITY
16:30 – 18:00
MS13.01: THE MEXTAG COLLABORATIVE
CROSS: IDENTIFYING THE GENETIC BASIS OF
MESOTHELIOMA
Scott Fisher1, Kimberley Burton1, Willem J. Lesterhuis1, Bruce
Robinson1, Grant Morahan2, Graeme Walker3 , Richard Lake1
School of Medicine and Pharmacology, National Centre for
Asbestos Related Diseases, The University of Western Australia,
Perth, WA, AUSTRALIA, 2Centre For Diabetes Research. The
University of Western Australia, Harry Perkins Institute of Medical
Research., Perth, WA, AUSTRALIA,3Drug Discovery Laboratory, QIMR Berghofer Medical Research Institute, Herston, QLD,
AUSTRALIA
1
the genetic heterogeneity of disease as well as complications of
environmental variation between cases and control groups (e.g.
amount of carcinogen exposure, effect of smoking and diet).
Thus, at least so far, the genes detected by GWAS have a minor
impact on disease risk and it is not known where in the disease
pathway they act, nor how they contribute to disease mechanisms. Here we outline a new strategy designed to rapidly
identify the genes associated with risk of mesothelioma. To define mesothelioma susceptibility and resistance loci, this study
combines the Collaborative Cross (CC) with our well-characterised MexTAg transgenic mouse model of mesothelioma. The CC
is a powerful mouse resource specifically developed to rapidly
identify genes associated with complex traits, while MexTAg
mice rapidly, uniformly and predictably develops mesothelioma,
but only after asbestos exposure.
Methods: 1: Generate and expose CC-MexTAg mice to
asbestos and assess mesothelioma development. The CC is a
collection of 100 recombinant inbred mouse lines covering over
90% of the common allelic diversity of the mouse species. The
genome sequences of each of the 100 CC lines are known. Each
CC line will be crossed with MexTAg mice and the resulting
CC-MexTAg progeny exposed to asbestos. Exposed mice will be
monitored for overall survival, the time to disease onset and the
time to disease progression. Disease will be confirmed by histological analysis. 2: Identify candidate modifier genes associated
with mesothelioma latency and time to progression. Using our
established informatics pipeline, we will rapidly identify alleles
that 1) protect mice against, or sensitize to mesothelioma, and
2) influence the pathology and course of the disease.3: Validate
the candidate modifier genes in the well‑defined Wittenoom
cohort of asbestos-exposed subjects. Human orthologues of
candidate genes will be validated in our mesothelioma GWAS
dataset.
Results: Breeding of CC-MexTAg progeny has begun and the
first batch of asbestos exposure experiments are underway.
Interim results will be presented at iMig 2016.
Conclusion: This study will provide insight into the genetic
factors underlying differences in mesothelioma development
after asbestos exposure. We aim to identify and validate modifier genes that will lead to an improved understanding of the
pathobiology of mesothelioma, identification of new druggable
targets and ultimately provide the necessary data for the development of diagnostic tests to assess individual risk of developing mesothelioma after asbestos exposure. Such tests would
identify the most at risk patients for further active monitoring
and treatment, while reassuring other patients that mesothelioma is unlikely to develop.
Keywords: mouse models, Collaborative Cross, MexTAg,
genetics of mesothelioma
Objectives: Mesothelioma development after asbestos
exposure is highly variable: some people do not develop the
cancer despite high level exposure for many years, while others
get disease with no known history of contact. There is good
evidence that at least part of the difference in susceptibility to
mesothelioma is genetic, but the genes involved remain mostly
unknown. Genome-wide association studies (GWAS) have been
used to try and identify mesothelioma susceptibility genes, but
the detection of significant associations in GWAS is hindered by
iMig2016.ORG
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ABSTRACT BOOK
MS13.02: TRANSLATIONAL CONTROL OF MPM:
ROLE OF EIF6 AND MICRORNAS IN METABOLISM
Stefania Oliveto1, Annarita Miluzio2, Pierluigi Gasparini3 ,
Roberta Alfieri2, Elisa Pesce2, Stefano Grosso4 , Sara Ricciardi2,
Daniela Brina2, Luciano Mutti5, Bruno Murer6 , Carlo M. Croce3 ,
Stefano Biffo7
Molecular Histology And Cell Growth Unit, INGM - Fondazione Istituto Nazionale Genetica Molecolare “Romeo ed Enrica
Invernizzi”, Milano, ITALY, 2INGM - Fondazione Istituto Nazionale
Genetica Molecolare “Romeo ed Enrica Invernizzi”, Milano,
ITALY, 3Department of Molecular Virology, Immunology and
Medical Genetics, Ohio State University Wexner Medical Center
and Comprehensive Cancer Center, Columbus, OH, UNITED
STATES OF AMERICA, 4MRC - Toxicology Unit, Leicester, UNITED
KINGDOM, 5School of Environment and Life Sciences, University
of Salford, Manchester, UNITED KINGDOM, 6Azienda ULSS 12
Veneziana, Venezia, ITALY, 7INGM - Fondazione Istituto Nazionale
Genetica Molecolare “Romeo ed Enrica Invernizzi” and University
of Milan, Milano, ITALY
1
Objectives: Translation is a cellular process finely regulated
during growth and development and it is deregulated in cancer
cells. Molecular mechanisms which control mRNA translation and the protein synthetic machinery are constituted by
steps potentially involved in tumorigenesis, pointing them as
novel druggable targets for cancer therapy. It has been shown
that eukaryotic Initiation Factor 6 (eIF6) is a limiting factor in
tumorigenesis, in vivo, regulating the availability of active 80S
subunit. A relationship between eIF6 activity and RISC complex
has been suggested, but remains controversial. We evaluated
the expression and activity of eIF6 in MPM, and its role in tumor
growth and metabolism. Moreover we investigated microRNAs
enriched on polysomes, and, among them, we focused on the
role and sublocalization of miR-24-3p in MPM cells.
Methods: The expression levels of eIF6 were measured by
IHC, WB analysis and qRT-PCR. 2D-electrophoresis has been
used to study the phosphorylation status of the protein.Cell
proliferation was analysed by MTT assay and FACS analysis was
used to determine cell cycle progression and apoptotis. Tumor
growth, in vivo, has been analysed by xenograft tumor technique. Methionine incorporation assay and polysomal profiles
were used to determine the translational status of cells.We
showed metabolism impairment in eIF6 knockdown cells using
lactate and ATP assays. Finally, we isolated RNA from polysomal profiles fractions of MPM cells and proceeded to microRNAs profiling. We also performed RNA seq analysis to identify
targets of miR-24-3p.
Results: We observe that Malignant Pleural Mesothelioma
(MPM), a tumor characterized by 100% lethality at two years
from diagnosis, exhibits high levels of eIF6 and differential
subcellular miRNAs distribution. We show that MPM contains
high levels of hyperphosphorylated eIF6 and that PKCβ inhibitor
Enzastaurin (Ely-Lilly) induces eIF6 dephosphorylation in
time-dependent manner. Treatment of mesothelioma cells, with
either Enzastaurin or shRNA for eIF6 affects cell growth, in vitro,
and causes reduced tumor growth and metastasis formation,
in vivo. Molecular analysis reveals that eIF6 manipulation
affects the metabolic status of malignant mesothelioma cells,
evidencing less glycolysis and less ATP content in cells depleted
for eIF6 or treated with Enzastaurin. Moreover, sucrose density
gradient analysis of MPM cells identified miRNAs in RNA
subpopulations: miRNAs distribution both in monosomes and
active polysomes is characterized by high variability in miRNAs
occupancy. We evidenced that polysome-bound miRNAs present a correlation with the cell cycle pathway and that miR-243p shows a significant polysomal localization. We found that in
spite of being upregulated in most MPM cell lines, miR-24-3p
displays a different expression between epitheliod and sarcomatous histosubtypes, and its inhibition has different effects.
Conclusion: We propose that eIF6 is necessary for Malignant
Mesothelioma growth, in vivo, and can be targeted by kinase
inhibitors and we suggest a new translational role of mir243p in MPM, taking in account its localization and cell specific
function. Cell specific miR-24-3p targets are characterized by
RNA sequencing analysis.
Keywords: Malignant Pleural Mesothelioma; Translation; eIF6;
phosphorylation; PRKCB; miRNA
MS13.03: BAP1 KNOCKOUT BY CRISPR-CAS9
GENOME EDITING IN MALIGNANT PLEURAL
MESOTHELIOMA CELL LINES FOR ISOGENIC
FUNCTIONAL STUDIES
Julija Hmeljak, Lee Spraggon, Marc Ladanyi
Human Oncology And Pathogenesis Program, Memorial Sloan
Kettering Cancer Center, New York, NY, UNITED STATES OF
AMERICA
Objectives: BAP1 loss is the third most prevalent genetic
alteration in malignant pleural mesothelioma (MPM). BAP1 is a
multifunctional deubiquitinase involved in chromatin dynamics,
DNA repair and cell cycle regulation. Current data suggest that
the role of BAP1 inactivation via mutation or loss in MPM is
complex. In order to elucidate the effect of BAP1 loss in MPM
cells, we used the CRISPR-Cas9 system to genetically ablate
BAP1 in BAP1 wild-type MPM cell lines.
Methods: HMeso (biphasic MPM) and VAMT (sarcomatoid
MPM) were transfected with a cocktail of three unique bifunctional CRISPR plasmids, each containing humanized Cas9
and a single guide RNA (gRNA) targeting the 5’ coding region
of BAP1. We isolated single HMeso BAP1-/- and VAMT BAP1/ clones, which were confirmed to be negative for BAP1 protein
expression and were screened at the genomic level to confirm
the correct gene editing by CRISPR-Cas9. Heterozygous partial
knockout clones (BAP1+/-) were also isolated and analyzed.
Results: Utilizing these newly developed isogenic cell lines,
we are performing extensive in vitro characterization, gene
expression profiling, and assessing response to radiation in the
context of growth delay and DNA damage.
Conclusion: These isogenic cell line models will provide a
powerful platform in which to further investigate the role of
BAP1 in MPM biology.
Keywords: BAP1, CRISPR, isogenic cell lines
iMig2016.ORG
58
ABSTRACT BOOK
Table 2: Effect of BAP1
MS13.04: BAP1 EXPRESSION AND IMPACT ON
TREATMENT OUTCOMES IN MALIGNANT PLEURAL
MESOTHELIOMA IN A PROSPECTIVE UK BASED
CLINICAL TRIAL
Neelam Kumar1, Krishna Kolluri1, Elaine Borg2, Elizabeth Sage3 ,
Zhi Zhou1, Mary Falzon2, Sam Janes1
Ucl Respiratory, Division of Medicine, University College London,
London, UNITED KINGDOM, 2University College London Hospital,
London, UNITED KINGDOM, 3University College London, London,
UNITED KINGDOM
1
Objectives: Genomic studies of malignant pleural mesothelioma (MPM) have identified frequent mutations in BRCA Associated Protein 1 (BAP1). BAP1 is a nuclear deubiquitinase with
important roles in regulating gene expression and DNA repair.
Previous studies have identified 100% correlation between
nuclear staining for BAP1 and wild type BAP1 status, pointing
to immunohistochemistry (IHC) as a reliable technique to
detect BAP1 molecular status. The objective of this study is to
assess BAP1 expression and infer molecular status using IHC
in a cohort of patients from a prospective UK based clinical
trial (MSO1 trial). Furthermore, we aim to evaluate the effect
of BAP1 status on treatment outcomes. This is the first assessment of BAP1 status in MPM in a UK patient cohort.
Methods: BAP1 expression was evaluated by IHC in 79 tumour
biopsies collected during the MSO1 trial by two consultant histopathologists. Cases were considered positive (wild type BAP1)
if strong nuclear staining was present throughout the tumour
and negative (mutant BAP1) if absent.
Results: Assessment of BAP1 expression was concordant in 77
of 79 cases (97%). BAP1 expression was negative in 66 of these
77 cases (86%). Patient characteristics are in Table 1 and the
effect of BAP1 expression on treatment outcomes in Table 2. Nuclear
BAP1 IHC
positive (N=11)
Nuclear
BAP1 IHC
negative (N=66)
p-value
Gender (M=male)
M: 100%
M: 91%
0.30
Median age at diagnosis (years)
69.5
66.0
0.94
Table 1: Clinical
characteristics
Epithelioid
82%
89%
Biphasic
18%
9%
Sarcomatoid
0%
3%
0.94
Treatment
Active symptom
control (ASC)
36%
36%
ASC + vinorelbine
36%
32%
ASC + mitomycin,
vinblastine,
cisplatin
27%
32%
p-value
All
21.0
23.3
0.22
Active symptom
control (ASC)
24.3
25.0
0.62
ASC + vinorelbine
12.8
22.8
0.11
ASC + mitomycin,
vinblastine, cisplatin
15.7
19.4
0.69
Conclusion: BAP1 expression was negative in 86% of MPM
tumours suggesting a high frequency of BAP1 mutations in this
UK cohort. No significant differences in clinical characteristics or outcomes were noted between cases with positive or
negative BAP1 expression overall. When analysed by treatment
subgroup, there was a trend towards a survival benefit in cases
with negative BAP1 expression (BAP1 mutants) in the ASC
plus vinorelbine arm, but no statistically significant difference
in outcomes within any treatment arm. We plan to further
validate our findings by correlating BAP1 expression directly
with BAP1 molecular status using laser capture microdissection
and sequencing.
Keywords: BAP1, mesothelioma
MS13.05: RELATIONSHIP OF PD-L1 EXPRESSION
AND PROGNOSIS IN EPITHELIOID MESOTHELIOMA
Tohru Tsujimura1, Tomoo Kudo1, Yoshiyasu Shinohara1, Ayuko
Sato2, Shigeki Shimizu2, Takashi Daimon2, Seiki Hasegawa2,
Takashi Nakano2
Department of Pathology, Hyogo College of Medicine, Nishinomiya, JAPAN, 2Hyogo College of Medicine, Nishinomiya, JAPAN
1
0.57
Histology
Nuclear
Nuclear BAP1
BAP1 IHC
IHC positive
negative
status on median
overall survival from
diagnosis (months)
Objectives: Malignant pleural mesothelioma (MPM) is a refractory tumor with poor prognosis. The most common histological
type of MPM is epithelioid mesothelioma (EM). Programmed
cell death-1 ligand 1 (PD-L1), which participates in immune
evasion of tumor cells, has been reported to be involved in
progression and poor prognosis in various human tumors. The
purpose of this study is to clarify the clinical/prognostic significance of PD-L1 in EM.
Methods: We examined immunohistochemically PD-L1 expression in tumor cells (TCs) and tumor-infiltrating immune cells
(ICs) of 42 EM patients who underwent extrapleural pneumonectomy. Associations of clinicopathological characteristics
with PD-L1 expression were examined with the use of Fisher’s
exact test. The relationship between PD-L1 expression and the
patient’s survival was examined as follows. Overall survival
curve was estimated by using the Kaplan-Meier method and
was compared with the use of the log-rank test. Hazard ratios
and their 95% confidence intervals were estimated with the use
of Cox’s proportional-hazards model.
iMig2016.ORG
59
ABSTRACT BOOK
Results: Thirty (71%) TCs and 16 (38%) ICs were positive for
PD-L1. Pathological stage (stage I/II vs III/IV) was significantly
associated with PD-L1 expression in ICs. Overall survival was
significantly shorter in EM patients with PD-L1-positive TCs
and/or PD-L1-positive ICs than those with PD-L1-negative TCs
and PD-L1-negative ICs. The 5-year survival rate was lower in
EM patients with PD-L1-positive TCs (20.6%) than those with
PD-L1-negative TCs (46.0%).
Conclusion: These results indicate that PD-L1 expression
in TCs and/or ICs of EM is associated with poor prognosis of
patients. PD-L1 expression may have important therapeutic
implications for the management of EM.
Keywords: PD-L1, epithelioid mesothelioma, prognosis
calculations were validated using literature searches and in
vitro based experiments. The comparison of model simulations
with genome wide microarray data demonstrated a significant
rate of correct predictions derived when compared to various
microarray profiles obtained from different cancel cells.
Conclusion: In summary, we show that the use of systems biology approaches to generate dynamic logical models provides
predictive value and better understanding of cancer development. Patient-specific microarray data will be integrated into
these simulations: by assigning initial node states to observed
patient-specific expression levels, personalised predictions will
be generated to identify mutation drivers/abrogated pathways
and novel druggable target. Finally, this approach will facilitate
the continual personalisation of MPM patients’ treatment based
on identifying shifts in signalling pathways that give rise to
resistance to therapy for a particular tumour.
Keywords: cancer, mesothelioma, p53, systems biology
MS13.06: SYSTEMS BIOLOGY APPROACHES
TOWARDS DEVELOPING PERSONALIZED
CANCER THERAPIES FOR MALIGNANT PLEURAL
MESOTHELIOMA (MPM)
MS14:RADIOTHERAPY
16:30 – 18:00
Michelle Hussain1, Alice Guazzelli1, Jean-Marc Schwartz2,
Luciano Mutti1, Marija Krstic-Demonacos1
School of Environment And Life Sciences, University of Salford,
Salford, UNITED KINGDOM, 2Faculty Of Life Sciences, University
of Manchester, Manchester, UNITED KINGDOM
1
Objectives: Malignant Pleural Mesothelioma (MPM) is an aggressive cancer with no effective treatments and poor prognosis. Chemotherapy is a common treatment used to treat cancer
patients, however, overall this treatment is often ineffective
highlighting the needs for novel approaches. The overall aim of
this research is to understand the MPM biology, the mechanism
of resistance to therapy and facilitate patients stratification
to improve survival. The knowledge of pathway changes will
facilitate patient stratification and result in personalised, more
effective therapeutic strategies. Numerous literature and large
datasets have been accumulated about cancer causes and
therapies, however systematic approaches with clear potential
for clinical applications are lacking.
Methods: Here we employ a systems biology approach including bioinformatics, text mining and logical modelling combined
with in vitro laboratory based experiments to analyse pathways
affected in MPM. One of the genes crucial for cancer development and treatment is the p53 tumor suppressor. Despite the
80,000 publications associated with the p53 in PubMed, the
details of p53 function are still unclear due to the complexity of
its interactions. Using a systematic approach we integrate this
vast amount of information by constructing a large-scale logical
model of the p53 interactome from databases and literature
information
Results: Our previously generated model containing 205 nodes
representing genes or proteins, DNA damage input, apoptosis
and senescence outputs, and 677 logical interactions was improved and adapted by addition of genes suggested as the drivers of MPM, to create a model consisting of 300 nodes and 860
interactions between them. Predictions from in silico knock-outs
mimicking mutations, steady state analysis and dependency
MS14.01: EXAMINING SIGNALING PATHWAY
CROSSTALK IN MESOTHELIOMA PDT AND RT
SENSITIVITY USING 2D AND NOVEL 3D TISSUE
CULTURE MODELS
Keith A. Cengel1, Sarah Hagan2, Edmund Moon3 , Charles B.
Simone1
Radiation Oncology, University of Pennsylvania, Philadelphia,
PA, UNITED STATES OF AMERICA, 2University of Pennsylvania,
Philadelphia, PA, UNITED STATES OF AMERICA, 3Division of Pulmonary, Allergy And Critical Care Medicine, Perelman School of
Medicine, University of Pennsylvania, Philadelphia, PA, UNITED
STATES OF AMERICA
1
Objectives: Photodynamic therapy (PDT) and external beam
radiotherapy (RT) have been used as adjuvant therapies directed at increasing local control in patients undergoing surgical
resection of malignant pleural mesothelioma (MPM). In patients
undergoing radical pleurectomy and PDT for MPM, we have
previously demonstrated that expression/activation of epidermal growth factor receptor (EGFR)/STAT3 signaling correlates
with increased pleural recurrence rates and decreased overall
survival. We have also shown that activation of STAT3 through
EGFR pathway activation mediates resistance of lung cancer
cells to both PDT and ionizing RT. Both EGFR and STAT3 are activated in the wound healing/inflammatory response to surgical
injury, and EGFR/STAT3 activation leads to increased cox-2 expression. Therefore, we hypothesized that STAT3 might mediate
crosstalk between inflammatory and growth factor signaling.
Methods: To begin to evaluate this hypothesis, we transfected
human mesothelioma cell lines derived from subjects enrolled
on tissue acquisition studies at Penn (EMM, EMP, Ren), with
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a pTRIPZ expression vector designed to allow doxycycline-induced STAT3 or EGFR shRNA expression. We then performed
clonogenic cell survival assays to assess the efficacy of PDT or
RT with or without inhibition of inflammatory signaling by celecoxib (cox-2 inhibitor). After PDT or RT, cell death was quantitated using calcein/ethidium bromide fluorescent live/dead cell
assays. Protein expression/signaling activation was measured
by Western blot analysis of tumor nodule protein extracts. All
assays and Western blot analyses were run in distinct biological
triplicates.
Results: These studies demonstrated that inhibition of EGFR/
STAT3 signaling pathways significantly enhance PDT-mediated
cellular cytotoxicity. The addition of celecoxib was found to
further enhance this effect. To evaluate the role of these pathways in a more clinically relevant in vitro model, we developed
a novel 3D tissue culture model of human MPM cells in which
shRNA expression vector transfected cells are implanted superficially on a matrigel pad and allowed to develop into tumor
nodules measuring 100-150 micrometers in diameter. Preliminary results from experiments using this tumor nodule/gene
expression assay system demonstrated significantly enhanced
cytotoxicity following PDT in the presence of EGFR/STAT3 pathway inhibition. Enhanced cytoxicity with EGFR/STAT3 inhibition
similarly was also seen following ionizing RT.
Conclusion: The results using 2D tissue culture demonstrate
that EGFR/STAT3 signaling pathway activation mediates
resistance of MPM cells and tumor nodules both to PDT and
RT. In addition, we have developed a novel 3D MPM nodule
model with the ability to selectively inhibit gene expression
using inducible shRNA and validated our findings from 2D tissue
culture. Further work is underway to develop heterotypic MPM
nodules using combinations of human fibroblasts, macrophages and lymphocytes to better mimic the complex cellular
interactions in MPM.
SYSTEMS Trial was the first prospective study of radiotherapy
in MPM to use validated outcome measures for pain. This multicentre, single arm phase II study of conventional dose palliative
radiotherapy (20Gy in 5#) recruited 40 patients in three centres
over 18 months and demonstrated that approximately one third
of patients experienced clinically meaningful improvements
in pain with minimal toxicity. It is hypothesised that increasing
the total radiation dose and the dose per fraction will result in a
greater proportion of patients experiencing a clinically significant pain response. The objective of the SYSTEMS-2 Study is to
compare a hypo-fractionated, dose escalated regime (36Gy in 6
fractions), with the standard dose (20Gy in 5 fractions).
Methods: This study aims to recruit 112 patients with a diagnosis of MPM, in whom radiotherapy is indicated for pain control.
Patients should have a worst pain score of >/= 4/10 (measured
by the Brief Pain Inventory, BPI) after analgesia optimisation.
Recruitment will take place in 8-10 centres across the UK.
Randomisation will occur after completion of radiotherapy
planning, but prior to the first treatment and plans must be acceptable for both dose/fractionation regimes. Doses to organs
at risk (e.g. spinal cord) can be minimised using advanced
planning techniques such as Intensity Modulated Radiotherapy (IMRT), however IMRT availability will not be an essential
requirement for participating centres. If there is concern regarding proximity of the tumour to an organ at risk, the final fraction
of the dose escalated arm can be omitted.
Keywords: 3D tissue culture models, photodynamic therapy,
radiation therapy, signal transduction
MS14.02: SYSTEMS-2:RANDOMISED PHASE II
TRIAL OF STANDARD VERSUS DOSE ESCALATED
RADIOTHERAPY FOR PAIN IN MALIGNANT
Miranda J. Ashton1, Anthony Chalmers2, Nicholas Macleod3 ,
Noelle O’Rourke3 , Barry Laird4
Beatson West of Scotland Cancer Centre, Glasgow, UNITED
KINGDOM, 2Institute of Cancer Sciences, University of Glasgow,
Glasgow, UNITED KINGDOM, 3Beatson West of Scotland Cancer,
Glasgow, UNITED KINGDOM, 4Edinburgh Cancer Research Centre, University of Edinburgh, Edinburgh, UNITED KINGDOM
1
Objectives: Pain is one of the most common symptoms
associated with malignant pleural mesothelioma (MPM) and is
often poorly responsive to analgesia. Palliative radiotherapy is
a recognised component of standard treatment for MPM-associated pain; however, there is no consensus on optimal dose,
fractionation or technique, and very little efficacy data. The
Results: Primary endpoint will be pain assessed at 5 weeks
measured by BPI, with a fall of >/=2 points on the BPI constituting a significant response. Secondary endpoints will be
assessed using validated tools and will include acute toxicities,
radiological response, overall survival and quality of life. Exploratory endpoints will include change in strong opioid use and
potential predictive and response biomarkers.
Conclusion: It is anticipated that this study will provide robust
and accurate symptom response data for palliative radiotherapy
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in MPM and help to establish the optimal dose and fractionation. This will guide clinical practice and should aid more
effective palliation for patients with MPM-associated pain.
Keywords: Pain, radiotherapy, palliation, dose-escalation
MS14.03: OVERCOMING RADIATION RESISTANCE
OF MESOTHELIOMA BY ACTIVATING TUMOUR
SPECIFIC CELL DEATH
Saurabh Dayal1, Lesley Gilmour1, Marcel Verheij2, Anthony
Chalmers3
University of Glasgow, Glasgow, UNITED KINGDOM, 2Netherlands Cancer Institute, Amsterdam, NETHERLANDS, 3Institute
of Cancer Sciences, University of Glasgow, Glasgow, UNITED
KINGDOM
1
Objectives: The incidence of mesothelioma is increasing and
current treatments are ineffective. While advances in technical
radiotherapy are increasing its potential clinical application,
intrinsic radiation resistance of mesothelioma remains an
important barrier. Defects in the apoptosis pathway are a likely
cause of radioresistance and treatment with Tumour necrosis
factor-Related Apoptosis Inducing Ligand (TRAIL) has potential
to overcome this.
Methods: Radiation and TRAIL (iso-leucine zippered form)
were tested alone and in combination in human mesothelioma
cell lines MSTO-211H, H2052 and H226 to determine effects
on cell viability, clonogenic survival and apoptosis (measured
by caspase-3/7 activity and Annexin-V/PI analysis). Synergy
was assessed by isobologram analysis. Mechanisms underlying interactions between radiation and TRAIL were probed by
measuring cell surface and total levels of the death receptors
DR4 and DR5 using immunoblotting, flow cytometry and immunofluorescence and by documenting effects of death receptor
knockdown and specific caspase inhibitors on apoptosis and
cell viability. TRAIL/radiation combinations were evaluated in
vivo using subcutaneous MSTO-211H xenografts.
Results: Radiation and TRAIL exhibit schedule-dependent
synergy. TRAIL treatment 24 hours after radiation was associated with significant increases in apoptosis and reductions in cell
viability and clonogenic survival in all cell lines tested. Consistent with this, radiation caused upregulation and externalisation
of DR4 & DR5 with maximum effects 24 hours after treatment.
We hypothesised that radiation induced DR4/5 upregulation
and externalisation enables activation of the extrinsic apoptotic
pathway by TRAIL. This was verified by showing that inhibition
of the extrinsic apoptotic pathway blocked the cytotoxic effects
of the radiation/TRAIL combination whereas inhibition of the
intrinsic pathway did not. Furthermore, siRNA knockdown of
DR5 abolished the radiosensitising effect of TRAIL. Effects of
radiation/TRAIL combinations in vivo will be presented.
Conclusion: Addition of TRAIL overcomes radioresistance
exhibited by mesothelioma cells by activating the extrinsic
apoptotic pathway. The synergistic combination of TRAIL and
radiation has therapeutic potential in mesothelioma.
Keywords: mesothelioma, TRAIL, apoptosis, radiotherapy
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ABSTRACT BOOK
MS14.04: SYNERGISTIC EFFECT OF LOCAL
RADIATION THERAPY AND IMMUNOTHERAPY IN A
MOUSE MODEL OF MPM
Luis De La Maza-Borja1, Matthew Wu1, Licun Wu1, Zhihong
Yun1, Marc De Perrot2
Thoracic Surgery Research, University Health Network, Toronto,
ON, CANADA, 2Thoracic Surgery, Toronto General Hospital and
Princess Margaret Cancer Center, Toronto, ON, CANADA
1
Objectives: Our group developed a new approach focusing on
Surgery for Mesothelioma After Radiation Therapy (SMART),
with encouraging results in a phase I/II clinical trial. We believe that radiation is important to achieving activation of the
immune system and may contribute to the benefits observed
in patients. Objective: To develop a mouse model to analyze
the immunogenic effect of Local Radiation Therapy (LRT), its
impact on immune cell recruitment and the effect of adjuvant
immunotherapy in the context of MPM. We hypothesized that
LRT administered to a tumor before surgery is immunogenic
and that adjuvant immunotherapy is beneficial and contributes
to tumor free survival.
Methods: Mice were inoculated subcutaneously in the flank
with the MPM cell line AB12 and AE17-OVA and were treated
with LRT in three 5Gy fractions. Radiation and untreated tumors
were analyzed for CD8+ T cell tumor infiltration. Furthermore,
tumor infiltrating lymphocytes (TILs) were analyzed with flow
cytometry using H-2Kb tetramer SIINFEKL as wells as activation markers. To assess the effect of adjuvant immunotherapy,
tumor bearing mice were treated with anti-PD1, anti PD-L1
and anti-CTLA-4 alone or in combination with LRT. To assess
protective memory, AE17-OVA tumor bearing mice were treated
with LRT and radical surgery and compared to radical surgery
alone. Cured mice were then rechallenged in the opposite flank
and tumor growth was followed.
Results: Tumor growth was decreased in mice treated with LRT
and showed a significant increase in the number of infiltrating
CD3+CD8+ cells compared to untreated tumors. Radiated
tumors also showed a greater proportion of tetramer specific
CD8+ T cells. Furthermore, infiltrating tetramer-specific CD8+
T cells showed significant upregulation of the activation marker
4-1BB and significant downregulation of the exhaustion marker
PD-1. Combination of LRT and immunotherapy showed an
important synergistic effect in mice treated with anti CTLA-4
and only a modest effect in mice treated with anti PD-1 and anti
PD-L1.In the rechallenge experiment, mice treated with LRT
and radical surgery showed significant deceleration in tumor
growth after rechallenge compared to radical surgery alone,
moreover, 3 out of 10 mice in the LRT and radical surgery group
completely rejected the tumor compared to 0 out of 10 mice in
the radical surgery group.
MS14.05: THE SMART TRIAL - AN RCT OF
PROPHYLACTIC RADIOTHERAPY IN PREVENTING
PROCEDURE TRACT METASTASES IN
MESOTHELIOMA
Nick Maskell1, Amelia O. Clive2
Academic Respiratory Unit, University of Bristol, Bristol, UNITED
KINGDOM, 2Respiratory Research Unit, North Bristol NHS Trust,
Bristol, UNITED KINGDOM
1
Objectives: The role of prophylactic radiotherapy to prevent
the development of procedure tract metastases (PTM) in
patients with malignant pleural mesothelioma has been subject
to intense debate. We examined its efficacy in preventing this
complication with a suitably powered, randomised controlled
trial.
Methods: We randomly allocated patients with histo-cytologically proven mesothelioma, who had undergone a large
bore pleural intervention in the previous 35 days to immediate
radiotherapy (21 Gray in 3 fractions) or deferred radiotherapy
(given if a PTM developed). Patients were followed up for 12
months. The primary outcome was the rate of PTM until death
or 12 months. Secondary outcomes included chest pain, quality
of life, analgesic requirements, health care utilisation and safety
(including radiotherapy toxicity).
Results: Two hundred and three patients were randomized
from 22 UK centres. The mean age was 71 (SD 8.1), 181/203
(89%) of patients were male, 142/203 (70%) had epithelioid
histology. The baseline characteristics of the treatment arms
were comparable. No significant difference was identified in the
rate of PTM between the immediate and deferred radiotherapy
groups (9/102 (8.8%) vs 16/101 (15.8%) respectively; OR 0.51
(0.19, 1.32); p =0.141). There was no difference identified in the
quality of life, chest pain, analgesia requirements or survival of
the two groups.
Conclusion: Among patients with mesothelioma, routine use
of prophylactic radiotherapy after large bore thoracic interventions does not confer benefits in terms of PTM rate, symptom
control or quality of life when compared to careful clinical
follow up and deferred radiotherapy should a PTM develop.
(Funded by a Research for patient benefit award from the National Institute of Health. ISRCTN72767336)
Keywords: randomised controlled trial, radiotherapy, mesothelioma
Conclusion: Local radiation therapy induced proliferation and
activation of specific anti-tumor CD8+ T cells. These specific
anti-tumor T cells may be responsible for rejection of the tumor
after rechallenge. Combination of radiation and immunotherapy showed a synergistic effect in controlling tumor growth.
Activation of the immune system secondary to LRT was further
improved with immunotherapeutic drugs.
Keywords: Radiation, CTLA4, Immunotherapy, Surgery
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ABSTRACT BOOK
MS15:MULTIMODALITY
16:30 – 18:00
MS15.01: COMBINED MODALITY TREATMENT
USING EXTRAPLEURAL PNEUMONECTOMY FOR
MALIGNANT PLEURAL MESOTHELIOMA: A SINGLE
CENTRE EXPERIENCE
Philippe Nafteux1, Stephanie Peeters2, Johnny Moons1, Yolande
Lievens3 , Melanie Dekeyser1, Christophe Dooms4 , Johan
Vansteenkiste4 , Paul De Leyn1, Kristiaan Nackaerts4
Thoracic Surgery, KU Leuven-University of Leuven, University
Hospitals Leuven, Leuven, BELGIUM, 2Radiotherapy, KU Leuven-University of Leuven, University Hospitals Leuven, Leuven,
BELGIUM, 3UGhent-University of Ghent, University Hospital
Ghent, Ghent, BELGIUM, 4Respiratory Diseases, KU Leuven-University of Leuven, University Hospitals Leuven, Leuven, BELGIUM
1
(n=52) were: 12,8%, 22.1% and 15.3% respectively. Subsequently, median disease-free survival of epithelial MPM pts was
better than for non-epithelial MPM pts, 21.5 and 13.5 months,
respectively (p=0.0037), even reaching 34.9 months for pN0
patients (n=33). Cox proportional hazards model showed a significantly lower DFS in non-epithelial MPM pts (HR=4.24, 95%
CI=1.81-9.98; p=0.001) and in node positive patients (pN1 pts:
HR=4.55, 95% CI=1.09-19.00; p=0.038 and pN2 pts: HR=2.57,
95% CI=1.06-6.23; p=0.036). None of the other examined
covariates (laterality, pT- nor R-status) reached significance.
Conclusion: This study demonstrated that CMT with EPP
for MPM pts is feasible and safe, with an acceptable surgical
mortality rate, in a tertiary referral centre setting. Careful
patient selection (staging and physical performance) is of
highest importance, given that only half of all ‘eligible’ pts will
finish CMT. Recurrence of MPM is mostly observed in pts with
non-epithelial MPM subtype and with nodal disease. Median
disease-free survival for epithelial MPM pts who complete CMT
looks promising, although unfortunately not yet validated in
proper randomised-controlled trials.
Keywords: radiotherapy, induction chemotherapy, extrapleural
pneumonectomy, combined modality treatment
Objectives: Combined modality treatment for MPM patients
(pts) remains a matter of debate, especially regarding the
choice of surgery between extrapleural pneumonectomy (EPP)
or pleural decortication (P/D).
Methods: Our combined modality treatment (CMT) protocol,
starting in 2003, consisted of induction chemotherapy (IC),
followed by EPP and thoracic radiotherapy (RT). Eligibility
criteria of candidates (inclusion criteria: age ≤ 70 years, WHO
≤1, medically fit for pneumonectomy, MPM staging cT2N2M0
or less for all histologic subtypes) were discussed at the weekly
multidisciplinary round. IC consisted of 3 cycles of cisplatin
(75mg/m² D1 q3wks) and pemetrexed (500mg/m² D1). If
non-progressive, EPP was performed followed by hemithoracic RT (most frequently intensity-modulated radiotherapy,
IMRT; dose 54Gy/1.8Gy ± boost). Survival was calculated from
histological confirmation of MPM diagnosis and analysed by
Kaplan-Meier.
Results: From March 2003 till December 2014, in total 197
MPM pts were discussed of which 97 pts started CMT. Histologic subtypes: epithelial (n=79), non-epithelial or mixed (n=18).
Clinical TNM staging IA/IB/II/III: 9/8/57/23 pts. After IC, 13 pts
had “progressive disease”, 5 pts were deemed irresectable and
3 pts refused surgery. Response rate of IC: CR/PR/SD/PD in
3%/30%/53%/14% of pts. A total of 76 pts underwent surgery:
EPP in 56 pts, exploratory thoracotomy in 20 pts (inoperable
chest wall invasion). Surgical (in-hospital) mortality was 3.6%
(2/56 pts). After EPP, 5 pts were not referred for RT because
of: unique kidney(1), postoperative empyema(1), multidisciplinary decision(1), SAKK 17/04 trial inclusion(2). Two pts did
not complete radiotherapy (bone metastases development and
intercurrent respiratory failure). Finally, 52 pts received CMT, of
which 47 pts trimodality and 5 pts IC + EPP. Only slightly more
epithelial MPM pts (43/79, 54%) completed CMT compared to
non-epithelial MPM pts (9/18, 50%). Intent-to-treat median survival (n=97) and median survival of those who fully completed
CMT (n=52) were 22.4 months and 33.1 months, respectively.
Intent-to-treat overall 5-year survival (n=97), overall and disease-free 5-year survival in MPM pts who fully completed CMT
MS15.02: OUTCOME OF TRIMODALITY THERAPY
INCLUDING INTRACAVITARY HYDROGEN
PEROXIDE TREATMENT IN MALIGNANT PLEURAL
MESOTHELIOMA
Mir A. Hoda1, Thomas Klikovits1, Viktoria Laszlo1, Clemens
Aigner2, Georg Lang1, Shahrokh Taghavi1, Sabine Zöchbauer-Müller3 , Karin Dieckmann4 , Balazs Dome1, Walter Klepetko1
Division of Thoracic Surgery, Medical University of Vienna, Vienna, AUSTRIA, 2Thoracic Surgery, Ruhrlandklinik - University of
Essen, Essen, GERMANY, 3Oncology, Medical University Vienna,
Vienna, AUSTRIA, 4Radiation Oncology, Medical University Vienna, Vienna, AUSTRIA
1
aggressive malignancy related to asbestos exposure. Trimodality therapy (TMT) involving macroscopic complete resection
either by extrapleural pneumonectomy (EPP) or pleurectomy/
decortication (P/D) with neoadjuvant or adjuvant chemotherapy and adjuvant radiotherapy is a widely accepted treatment
protocol with curative intent. Recently a combination of TMT
with intracavitary treatment strategies has been reported to be
beneficial. In this study we investigated the impact of TMT in
combination with intraoperative hydrogen peroxide treatment in
patients with MPM.
Methods: All MPM patients who were referred for surgical
therapy within a TMT protocol between 2000 and 2012 were
enrolled in this study. Data was collected retrospectively until
2005 and prospectively from 2006 from patients who underwent at least 3 cycles of induction chemotherapy followed
by EPP and intraoperative hydrogen peroxide treatment and
postoperative intensity modulated radiotherapy up to 58 Grays.
In order to investigate the effect of hydrogen peroxide in vitro on
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ABSTRACT BOOK
MPM cell growth, MPM cell lines (N=10) were treated with different concentrations of hydrogen peroxide and sulforhodamine
B and clonogenic assays were performed.
Results: 30 patients completed TMT and intraoperative hydrogen peroxide treatment during the observation period. There
were 24 males and 6 female with a mean age of 61 years at the
time of diagnosis. Median follow-up was 18.5 months. Histological subtypes were epitheliod MPM in 23, biphasic MPM in 6 and
sarcomatiod MPM in 1 patient. Eighteen patients were in late
stages whereas 12 patients were in early stages at the time of
diagnosis. Four (13.3 %) patients experienced major postoperative complications. These complications requiring re-thoracotomy were: bleeding , patch rupture, bronchopleural fistula.
Overall median survival (Kaplan-Meier) was 31 months (95%
confidence interval: 12–48 months). One-year survival was
80%, 2-year survival was 55% and 3-year survival was 45%. 30
day mortality was nill. In vitro, hydrogen peroxide inhibited the
growth of MPM cells in short- and long-term viability assays at
already low concentrations after 5 minutes of treatment.
Conclusion: In our experience, EPP in combination with intraoperative hydrogen peroxide treatment within a TMT protocol is
a well-tolerated and feasible treatment approach. Compared to
reported classical TMT protocols trials, morbidity and perioperative mortality rates are lower and median survival is equal or
better. Furthermore we were able to show that hydrogen peroxide inhibited MPM cell growth in vitro after short exposure.
Keywords: Surgery, multimodality treatment, intraoperative
treatment
MS15.03: CYTOREDUCTIVE SURGERY
AND HYPERTHERMIC INTRATHORACIC
CHEMOTHERAPY FOR TREATMENT OF PLEURAL
MESOTHELIOMA: A 10-YEAR EXPERIENCE
Pietro Bertoglio1, Marcello C. Ambrogi1, Antonio Chella2,
Vittorio Aprile1, Stylianos Korasidis1, Marco Lucchi1, Paolo Dini1,
Olivia Fanucchi1, Alfredo Mussi1
University Hospital Of Pisa, Division of Thoracic Surgery, Pisa,
ITALY, 2University Hospital Of Pisa, Division of Pneumology, Pisa,
ITALY
1
Objectives: The primary end-point of this retrospective 10-year
study was to evaluate safety and feasibility of cytoreductive
surgery followed by hyperthermic intrathoracic chemotherapy
(HITHOC) perfusion for treatment of Malignant Pleural Mesothelioma (MPM). The secondary end-point was survival, disease
free interval and analysis of possible risk-factors.
Methods: The outcomes of all patients with MPM who underwent cytoreductive surgery followed by HITHOC, during the
period 2005-2014, were analyzed. All patients were evaluated in
a multidisciplinary setting. Selection criteria were histologically-proven epithelioid or biphasic MPM; clinical IMIG stage I-III
disease; age 18-75 years; performance status < 3 according to
Eastern Cooperative Oncology Group; absence of cardiac, neuropathic or renal disease; adequate medullary reserve; no con-
comitant infection; no pregnancy status. Surgery consisted of
resection of parietal and mediastinal pleura plus gross debulking of any disease on the diaphragm and the pericardium, which
were routinely spared. After thoracotomy closure, HITHOC was
run by the mean of the chest drains using a dedicated perfusion machine: Cisplatin (80 mg/m2) and Epirubicin (25mg/m2)
perfusion lasted for 60 minutes at a target temperature of 42°C.
Intra-operative and post-operative morbidity and mortality were
recorded. All patients received at least three cycles of adjuvant
chemotherapy, while prophylactic radiotherapy was administered according to the choice of the referring oncologist. Survival analysis (calculated from the day of surgery) was performed
only on patients with a minimum follow-up of 12 months.
Results: Among 49 patients, 41 were male. Median age was 68
years (35-76). Histology was epithelioid in 43 cases. Pathological stage I, II, III and IV was presented in 12, 14, 20 and 3 cases
respectively. No intraoperative complications occurred and all
procedures were successfully completed. No 30- and 90-day
mortality were noticed. Morbidity consisted in anemia requiring
blood transfusion in 11 cases (22,45%), prolonged air-leak (>5
days) in 4 cases (8,16%), wound dehiscence in 3 cases (6,12%)
and one (2,04%) postoperative empyema, treated and resolved
with medical therapy. Median hospital stay was 8 days (5-45).
At a mean follow up period of 26 months, 12 patients were still
alive and 4 of them have no clinical or radiological signs of
disease. Actuarial median overall survival was 22 months and
a 1, 2 and 5 year survival accounted for 79,6, 43,3 and 13,7%
respectively. Median disease free survival was 12 months after
surgery. Age over 65 years and sex did not showed to be significantly related to the survival, while biphasic histology had a
significant worse prognosis compared to epithelioid (p=0,026).
Stage were a predictor of prognosis: patients with stage I had a
median survival of 46 months (p<0,001) and a median DFI of 18
months (p=0,004).
Conclusion: Cytoreductive surgery associated to HITHOC and
adjuvant chemotherapy is feasible and safe, with no mortality
and low morbidity, allowing a good control of the disease and
acceptable outcomes; its low invasiveness allows to extend
indication for surgery also to patients who might not be fit for
more invasive and detrimental procedures. Larger controlled
studies are needed to confirm our promising results.
Keywords: Malignant pleural mesothelioma, hyperthermic
intrathoracic chemotherapy, cytoreductive surgery, partial
pleurectomy
MS15.04: RADICAL PLEURECTOMY
CHEMOPERFUSION FOR THE TREATMENT OF
Laura V. Klotz, Michael Lindner, Jürgen Sklarek, Rudolf A. Hatz
Lungenfachklinik Gauting, Center for Thoracic Surgery Munich,
Gauting, GERMANY
Objectives: Radical pleurectomy and decortication (P/D) of
the parietal and visceral pleura represents a surgical treatment
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ABSTRACT BOOK
approach for patients suffering from malignant pleural mesothelioma (MPM) without the possibility to undergo pleuropneumonectomy. For several years, we combine this procedure with
hyperthermic intrathoracic chemoperfusion (HITHOC).
Methods: Between 2009 and 2014, 70 patients with malignant pleural mesothelioma underwent radical P/D followed
by HITHOC. Patients’ characteristics, tumor stage, intra- and
postoperative complications and overall survival were evaluated
retrospectively.
Results: The median patient age was 68 years. Sarcomatoid
compared to epitheloid MPM showed significant difference
in median survival after P/D and HITHOC with 9,2 months for
sarcomatoid and 18 months for epitheloid subtype. Concerning
epitheloid MPM, median survival was significantly better for
patients macroscopic complete resection (MCR) in contrast
to patients after macroscopic incomplete resection with 34,4
months versus 12,3 months.
Conclusion: Radical P/D in combination with HITHOC is a
promising treatment option for selected patients with epitheloid
MPM. Macroscopic complete resection is the most important
prognostic factor for survival.
events, hematological and renal toxicity were monitored using
CTCAE grading. Quality of life was assessed with the SF36v2
questionnaire, physical component summary (PCS) score and
mental component summary (MCS) score and were compared
to pre-treatment values using the Wilcoxon signed rank test.
Results: No dose limiting toxicity was observed. Major morbidity was observed in 4 patients (33%). 30day- and 90day-mortality was 0%. The median serum AUC0-24 in the highest dose
level group reached 23 h*µg/g even after induction chemotherapy, which is still below the suggested renal toxicity risk level,
25 h*µg/g [Royer 2008]. Local tissue cisplatin concentration
at 90 minutes varied from 12-133 (median: 36.5 µg/g). Serial
chest wall biopsies of 2 patients showed cisplatin at day 74 and
204 after application. PCS score was significantly decreased
initially but returned to baseline values at 8 months after application. The MCS score was never significantly different from
pretreatment evaluation. The median follow-up after surgery
was 17 months (range 11 – 36 months). Median freedom from
recurrence was 8 months (95% confidence interval (CI): 4-12
months). In one patient with IMIG stage I, no sign of relapse was
observed at 16 months after treatment (44 mg/m2 BSA). Median
overall survival was 21 months (95% CI: 14-28).
Keywords: pleurectomy, Malignant pleural mesothelioma,
Surgery, hyperthermic chemoperfusion
MS15.05: INTRACAVITARY CISPLATIN-FIBRIN
APPLICATION FOLLOWING RESECTION OF
MESOTHELIOMA
Isabelle Opitz1, Olivia Lauk1, Mayura Meerang1, Martina Friess1,
Michaela B. Kirschner1, Guillaume Wuilleret1, Cordelia Bommeli1, Alexander Jetter2, Beat Aeschlimann3 , Detlef Guenther3 , Rolf
Stahel4 , Walter Weder1
SWITZERLAND, 2Department of Clinical Pharmacology And Toxicology, University Hospital Zurich, Zurich, SWITZERLAND, 3Department of Chemistry And Applied Biosciences And Laboratory
of Inorganic Chemistry, ETH Zurich, Zurich, SWITZERLAND, 4Clinic For Oncology, University Hospital Zurich, Zurich, SWITZERLAND
1
Objectives: Early local tumor relapse is very common after
resection of malignant pleural mesothelioma (MPM). Intracavitary chemotherapy after tumor resection might improve local
tumor control. The present clinical trial assesses intracavitary
cisplatin-fibrin application after pleurectomy/decortication
(P/D) for MPM patients.
Methods: Twelve patients (75% IMIG stage III + IV) with a
median age of 65 years underwent intracavitary cisplatin –fibrin
application after P/D at 4 different dose levels of cisplatin
(11, 22, 33 and 44mg/m2). Eight patients were previously
treated with intravenous cisplatin/pemetrexed (100 mg/m2).
To evaluate cisplatin pharmacokinetics, blood and chest wall
tissue samples were taken at several time points. Cisplatin
levels were measured by inductively coupled plasma sector
field mass spectrometry. Besides adverse and serious adverse
Figure 1: Serum cisplatin AUC (A), tissue/fibrin cisplatin concen-
tration in serial biopsies from 2 patients (B).
Conclusion: The administration of intracavitary cisplatin-fibrin
as high as 44mg/m2BSA is safe after (e)P/D, also in combination with induction chemotherapy. Tissue cisplatin concentration was cytotoxic whereas no dose limiting toxicity due to
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systemic distribution was detected. Quality of life is normalized
8 months after treatment. A confirmation of the safety and
evaluation of efficacy of the highest dosage, 44 mg/m2BSA, in a
phase II trial is ongoing.
Keywords: intracavitary treatment, phase I clinical trial,
cisplatin
MS15.06: PLEURECTOMY / DECORTICATION AND
INTRAOPERATIVE INTRAPLEURAL HYPERTHERMIC
CDDP PERFUSION FOR MALIGNANT PLEURAL
MESOTHELIOMA
were not reached in P/D group and 17 months in EPP group.
Two and 3-year overall survivals were 90% and 68% in P/D
group, respectively, and 30% and 30% in EPP group, respectively. Median disease free survivals were not reached in P/D
group and 12 months in EPP group. Two and 3-year disease free
survivals were 57% and 57% in P/D group, respectively, and
30% and 20% in EPP group, respectively. First relapse sites
indicated that all 4 relapses were locoregional in P/D group,
while 6 of 7 relapses were distant and the other was both local
and distant in EPP group.
Conclusion: P/D and intraoperative hyperthermic CDDP
perfusion followed by systemic chemotherapy for resectable
MPM was promising. Further study with more patient accrual is
warranted.
Keywords: pleurectomy / decortication, hyperthermia, intrapleural chemotherapy, multimodality treatment
Kenichi Okubo, Hironori Ishibashi, Masashi Kobayashi, Chihiro
Takasaki, Sachiko Kumazawa
Thoracic Surgery, Tokyo Medical and Dental University, Tokyo,
JAPAN
Objectives: The role of surgery in the treatment for resectable
MPM is considered to be macroscopic complete resection.
We examined our results of multimodality treatment including
extended pleurectomy/decortication (P/D) for resectable MPM.
MS15.07: HIPEC AS A TREATMENT FOR
MALIGNANT PERITONEAL MESOTHELIOMA: ARE
WE THERE YET?
Methods: We have performed a treatment-protocol consisting
of P/D with intraoperative hyperthermic intrapleural CDDP
perfusion (42℃, CDDP 80mg/m2 in saline 2L, 1hr) and postoperative chemotherapy (CDDP+PEM, 4 cycles) for patients with
MPM (cT1-3N0-2M0) since 2010. Our indication of P/D-protocol
was patients with MPM who were intolerable for EPP until 2013,
and all patients with MPM after 2014. There were 10 men and
2 women with a mean age of 66.9 years (55-76 years). Five
patients had a right-side disease and 7 had a left-side disease.
Histological subtypes were epithelioid in 9, biphasic in 2, and
desmoplastic in 1, and pathological stagings were stage I in 4,
stage II in 1, stage III in 6, and stage IV in 1. Eleven patients underwent a P/D and hyperthermic chemoperfusion initially, and
one patient with secondary nephrotic syndrome received five
cycles of preoperative chemotherapy. All parietal and visceral
pleura were removed, and pericardium and/or diaphragm were
resected when necessary. Concentrations of platinum in the
perfusate were examined before and after perfusion in 7 patients. Acute surgical outcome, survivals and relapse patterns
in P/D group were examined, and compared with 10 patients
(EPP group) who received trimodality treatment consisting of
extrapleural pneumonectomy, chemotherapy, and intensity
modulated radiation therapy for entire hemithorax (50Gy) in our
institute during 2010-2013.
Gleneara E. Bates, Yaakov Bressler, Robert N. Taub
Results: All patients obtained macroscopic complete resection in P/D and EPP groups. All but one in P/D group received
multiple cycles of chemotherapy, and all in EPP group completed the trimodality treatment. Operation time was longer in
P/D group; however, there were no differences in ICU stays or
hospitalizations. Ten patients in P/D group and 7 patients in
EPP group experienced postoperative complications; however,
there were no operative mortality. EPP group suffered from
cardiac complications and P/D group had prolonged airleak.
Concentrations of platinum showed that 29% of CDDP was left
in the pleural cavity after the chemoperfusion. Median survivals
Objectives: Intraoperative hyperthermic intraperitoneal chemotherapy (HIPEC) is now advocated by numerous experienced
mesothelioma specialists as part of the standard of care for
malignant peritoneal mesothelioma (MPM).The earliest studies
with use of hyperthermia alone and intraperitoneal chemotherapy alone in humans were conducted in the 1970’s. Since, there
has been little systematic research conducted on how these
two treatments may be optimally used in combination.This
study investigates the largest single-institution cohort of MPM
patients treated with surgical cytoreduction and HIPEC guided
by a medical oncologist.
Methods: Kaplan-Meier curves and univariate cox proportional
hazards model were used to estimate survival and significant
treatment and prognosis factors for 195 patients who underwent cytoreductive surgery and or HIPEC treatment between
1995–2014; patients were not excluded for bicavity disease or
for unresectable disease.
Results: The median survival time was 3.21 years with (95%
CI: 2.38-5.53), with median follow-up of 3.44 years (SD=3.4,
minimum=0.014 and maximum=16.752) years from first operation. The mean age at diagnosis was 54.8 years [HR: 1.027
(95% CI: 1.012-1.042)] with 111 men (57%) and 84 women
(43.1), with female gender having favorable survival [HR: 0.442
(95% CI: 0.296-0.659)]. Asbestos exposure was reported in 77
patients (39.5%) with n=80 (41.0%) having no known asbestos
exposure and no documented exposure in 38 patients (19.5%).
Majority of patients had epithelioid histology (n=16, 82.6%),
with the remainder biphasic/sarcomatoid (n=34, 17.4%), with
increased risk of death with non-epithelioid histology [HR:
2.46(95%CI 1.59-3.82)], (P=0.001). Of the 195 patients who
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received cytoreductive surgery, 71 (36.1%) received 1 HIPEC
treatment, while 124 (63.9%) received 2 HIPEC treatments,
with completion of protocol having an associated favorable
prognosis [HR: 0.161 (95%CI 0.109-0.237)] (p= 0.001). Of those
who received a full treatment course of cytoreductive surgery
and 2 HIPEC treatments, 66 (33.8% CI: 95%) were alive at the
median follow-up.
Conclusion: This large cohort illustrates that CRS and HIPEC
may be an effective treatment for MPM. While the treatment
continues to evolve, the future of HIPEC for MPM will depend
largely on monitored multi-institutional collaborative randomized clinical trials to demonstrate whether HIPEC is the optimal
treatment for MPM. There is much variation of technique for
HIPEC among medical centers – parameters of such operations
include: open or closed perfusion, chemotherapeutic agents,
concentrations of agents, temperature range, and exposure
time. Standardizing HIPEC for MPM patients would be an
important contribution to the continued advancement of MPM
treatments.
Keywords: Peritoneal, Survival, HIPEC, mesothelioma
MS16: NOVEL IMMUNE STRATEGIES
16:30 – 18:00
Technologies in the pIDT.SMART cloning plasmid. It was then
subcloned into the previously described MigR1 retroviral vectors containing the CARs against human mesothelin (mesoCAR)
(Riese et al., 2013) and murine fibroblast activating protein
(FAPCAR) (Wang et al., 2014). Primary murine T cells were isolated and transduced with these retroviral particles. The mesoCAR and RIAD constructs were also subcloned into a lentiviral
vector used to transduce human T cells undergoing anti-CD3/
CD28 bead activation. CAR vs. CAR-RIAD T cells were tested
for tumor lytic and cytokine secretion function in the presence/
absence of PGE2 and adenosine. These T cells were also compared in murine models of established solid flank tumors.
Results: After exposure to PGE2 or adenosine in vitro, CAR-RIAD T cells showed increased TCR signaling, more cytokine
release, and enhanced killing of tumor cells compared to CAR
T cells. When injected into tumor-bearing mice, murine and
human CAR-RIAD T cells demonstrated enhanced anti-tumor
efficacy compared to CAR T cells due to increased T cell infiltration of established tumors, and attenuated tumor-induced
hypofunction. Subsequent in vitro assays showed that both
mouse and human CAR-RIAD cells migrated more efficiently
than CAR cells in response to the chemokine CXCL10 and also
adhered better to various matrices.
Conclusion: Our data therefore show that the addition of the
RIAD peptide to adoptively transferred CAR T cells augments
their efficacy by increasing their effector function and by
improving trafficking into tumor sites. This treatment strategy
should therefore be considered for clinical application in treating solid tumors.
Keywords: T cells, tumor microenvironment, Immunotherapy
MS16.01: BLOCKADE OF PROTEIN KINASE A (PKA)
LOCALIZATION AUGMENTS TRAFFICKING AND
ANTI-TUMOR EFFICACY OF CAR T CELLS
Edmund Moon1, Kheng Newick1, Shaun O’Brien2, Jing Sun2,
Albert Lo2, Steve Maceyko2, Steven Albelda2
Pulmonary, Allergy, And Critical Care, University of Pennsylvania, Philadephia, PA, UNITED STATES OF AMERICA, 2University of
Pennsylvania, Philadelphia, PA, UNITED STATES OF AMERICA
1
Objectives: In recent years, the adoptive transfer of T cells
transfected with chimeric antigen receptor (CAR) genes has
shown great promise in treating blood-borne tumors. However,
in the case of solid tumors like mesothelioma, many factors
exist that eventually render these CAR T cells ineffective. Two
such factors include the presence of immunosuppressive
mediators (such as prostaglandin E2 (PGE2) and adenosine),
and poor T cell trafficking. Since PGE2 and adenosine activate
protein kinase A (PKA), which then inhibits T cell receptor (TCR)
activation, we generated CAR T cells that expressed a small
peptide called the “regulatory subunit I anchoring disruptor”
(RIAD) that inhibits the association of protein kinase A (PKA)
with ezrin; this interaction is required for the negative effects of
PKA on TCR activation. We hypothesized that cloning the RIAD
transgene into T cells expressing CARs would enhance their
function within the tumor microenvironment and result in superior tumoricidal ability as compared to T cells with CAR alone.
Methods: The RIAD construct into which myc and ddk (FLAG)
tags were incorporated, was synthesized by Integrated DNA
MS16.02: CHARACTERISING T-CELL RESPONSES
AGAINST MUTATED MESOTHELIOMA NEOANTIGENS
Jonathan Chee1, Shaokang Ma1, Jenette Creaney1, Bruce
Robinson2
School of Medicine and Pharmacology, National Centre Of
Asbestos Related Diseases, University of Western Australia,
Nedlands, AUSTRALIA, 2School of Medicine and Pharmacology,
National Centre Of Asbestos Related Diseases, University of Western Australia, Nedlands, WA, AUSTRALIA
1
Objectives: A feature of carcinogen-induced cancers is the accumulation of mutations in the cancer cell. Cytotoxic T lymphocytes (CTLs) respond to mutated proteins (neo-antigens), and in
some experimental models, boosting CTL responses specifically against neo-antigens can cause tumour regression. High
throughput sequencing such as RNA and exome sequencing has
allowed us to interrogate mutations from different cancers. Application of algorithms on sequencing data allows us to predict
mutated proteins in a cancer that can be potentially recognised
by host immune cells such as cytotoxic CD8 T cells (CTLs).
We have sequenced mouse mesothelioma (AB1/AB1-HA),
and described the first mutated mesothelioma neo-antigens.
Furthermore, we have demonstrated immune responses against
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one of the predicted neo-antigens: mutated Uqcrc2 (Uq2) in
mesothelioma bearing animals. The objective of this study was
to characterize further characterize T cell responses against
neo-antigens in tumour bearing animals after different treatments that cause tumour regression in AB1 model. We hypothesize that treatments will increase the strength of responses to
‘existing’ neo-antigens such as Uq2, and could also lead to the
detection of responses against other neo-antigens that would
otherwise be undetectable without treatment.
aims to induce an effective in vivo anti-tumor T-cell response.
These T cells, however, are strongly suppressed by tumor associated macrophages (TAMs) within the tumor, thereby preventing an effective anti-tumor immune response from occurring.
TAMs are known to be short-lived cells and are highly dependent on signaling via the colony stimulating factor 1 receptor
(CSF1R). We investigated whether the efficacy of DC-therapy
could be enhanced by depleting TAMs through CSF1R-blockade
in a mesothelioma mouse model.
Methods: Mice were inoculated subcutaneously with mesothelioma cell lines (AB1-HA). AB1-HA bearing animals were treated
either with anti-CTLA4 antibody (checkpoint blockade immunotherapy), anti-GITR antibody (Treg modulation), gemcitabine
(chemotherapy), neo-antigen vaccinations or depleted of Tregs
(removal of immunosuppression). We tested T cell responses
at the draining lymph nodes and tumours against predicted
neo-antigens using different immunoassays such as IFNg ELISPOT, pMHC tetramers and flow cytometry.
Methods: Four groups of six female wildtype Balb/c mice were
injected intraperitoneally with a syngeneic mesothelioma cell
line (AB1) on day 0, followed by suboptimal treatment with
DC-therapy or PBS on day 10. DCs were cultured from wild-type
Balb/c bone marrow cells, loaded with an AB1-cell line lysate
and matured using a TLR9-agonist (CpG). Concurrently, mice
were fed from day 10 onwards either with a CSF1R-inhibitor in
chow (PLX3397) or control chow. Blood was obtained on day
15 and analyzed by flow cytometry to investigate peripheral
immune activation. Mice were sacrificed when they presented
with signs of severe illness or at the end of the experiment, 120
days post-tumor injection. End-stage tumors were investigated
using flow cytometry and immunohistochemistry to determine
the local immune composition.
Results: The frequency of T cells specific for a single neo-antigen (Uq2) is low (<0.1%). ELISPOT was the only immunoassay
tested that could detect responses against neo-antigens. Conventional pMHC staining and flow cytometry was not sensitive
enough to detect a positive signal for low frequency T cells.
Using the ELISPOT assay, we demonstrate the frequency of T
cell responses to neo-antigen Uq2 was highly variable between
mice after treatment, and showed a general trend of increase
after checkpoint blockade immunotherapy, chemotherapy or
Treg removal (compared to non-treated mice). Treatments did
not increase T cell responses to 30 other predicted neo-antigens.
Conclusion: The frequencies of neo-antigen specific T cells
are low and restricted to only one or a few mutated neo-antigens. For effective translation, e.g. using neo-antigen vaccines,
assays with increased sensitivities, or an additional T cell
expansion step must be used to detect and phenotype neo-antigen specific T cells. We detected an increase in neo-antigen T
cell responses after some treatments. Current studies include a
detailed analysis of Uq2 and other response, including whether
Uq2 or other neo-antigen responses are essential for an anti-tumour immune response.
Keywords: Immunotherapy, tumour neo-antigens, T cells,
tumour immunology
MS16.03: CSF1R-BLOCKADE SYNERGIZES
WITH DENDRITIC CELL IMMUNOTHERAPY IN A
MESOTHELIOMA PRECLINICAL MODEL
Floris Dammeijer, Lysanne Lievense, Menno Van Nimwegen,
Koen Bezemer, Margaretha Kaijen-Lambers, Rudi Hendriks,
Joost Hegmans, Joachim Aerts
Results: Analysis of peripheral blood from day 15 during
CSF1R therapy revealed no significant changes in CD4+ and
CD8+ T-cell numbers and no additional increase in proliferation
(Ki-67+) or effector markers (KLRG1) compared to DC-therapy
alone. However, Ly6G+ granulocytes and Ly6C+ monocytes
were decreased in numbers, accompanied by an increase in
immature myeloid cells defined by a Ly6C/Ly6G intermediate
phenotype. There was an increase in survival of mice treated
with the combination therapy as 50% of mice survived for the
duration of the experiment. In contrast, only one mouse survived when treated only with DC-therapy, and all mice had to be
sacrificed in the CSF1R-monotherapy and PBS control groups.
End stage tumors revealed a decrease in TAMs when mice were
treated with the CSF1R-inhibitor, however, mice that progressed
during treatment had surprisingly rare CD8-T-cell infiltrates. Coincidentally, these tumors were populated by dense infiltrates of
GR1+ cells
Conclusion: TAMs represent an abundant population in mesothelioma tissue. Using a CSF1R-inhibitor, we show that TAMs
can be selectively depleted in a mesothelioma mouse model,
and that CSFR1-blockade synergizes with DC-therapy. A subset
of mice progressed during CSFR1-blockade, a finding which
could be explained by the infiltration of GR1+ cells, indicating
a possible redundancy between these cells and TAMs in the
tumor. These findings pave the way for further experiments investigating the benefits of disabling local immune suppression
together with potent T-cell induction by DC-therapy in malignant mesothelioma.
Keywords: Tumor associated macrophages, Dendritic Cell
Immunotherapy, Malignant mesothelioma, Combination Immunotherapy
Dept. Of Pulmonary Medicine, Erasmus Medical Center, Rotterdam, NETHERLANDS
Objectives: Dendritic cell (DC)-based immunotherapy is a
promising treatment for malignant mesothelioma. DC-therapy
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MS16.04: PROSTAGLANDIN E2-CAMP-ADENOSINE
NEXUS IN MESOTHELIOMA; SELF AMPLIFYING
IMMUNOSUPPRESSION?
MS16.05: HETEROTYPIC 3D SPHEROID TUMOUR
MODELING OF MESOTHELIOMA TO DETERMINE
IMMUNE RESISTANCE
Zsuzsanna Tabi
Saly Al-Taei, Josephine Salimu, Zsuzsanna Tabi
Division of Cancer And Genetics, Cardiff University, Cardiff,
UNITED KINGDOM
Division of Cancer And Genetics, Cardiff University, Cardiff,
UNITED KINGDOM
Objectives: ATP-release by damaged or dying cells represents
a danger signal that contributes to the initiation of immune
responses. On the other hand, the ATP-metabolite adenosine is
well known for its extensive immunosuppressive effects and direct support of tumour growth. Cell surface expressed ectonucleotidases hydrolyse ATP in a sequential manner: CD39 hydrolyses ATP into ADP and 5’-AMP while CD73 hydrolyses AMP into
adenosine. CD73 can be expressed by malignant cells, while
CD39 by tumour stroma. Due to the combined activity of these
two enzymes, adenosine levels are often elevated in the tumour
tissue. Circulating human monocytes in healthy donors are
CD39-positive but do not express CD73. We observed recently
that monocytes in mesothelioma-associated pleural exudate
express both CD39 and CD73. The aim of this study is to reveal
the regulation and relevance of monocytic CD73 expression in
mesothelioma.
Objectives: Tumour modelling in vitro in the form of 3D spheroid cell culture provides a more accurate platform for assessing
the benefit of potential new therapeutics, when compared to 2D
culture. However, these homotypic tumour cell spheres still fail
to capture the complexity of tumour tissue. We have an ongoing
project to develop a complex 3D heterotypic tumour spheroid
model incorporating the main tumour components; tumour,
stromal and endothelial cells to more accurately mimic tumour
tissue in vitro. This model will enable us to study the susceptibility of mesothelioma tumours to immune cell killing. We predict
that the inclusion of fibroblasts and endothelial cells alongside
the tumour will provide marked protection of tumour cells from
T cell mediated killing.
Methods: CD14+ myeloid cells in the pleural effusion and
peripheral blood of patients and healthy donors were phenotyped by flow cytometry for the expression of CD39 and CD73.
Healthy donor CD14+ cells were separated and exposed to the
cell free fraction of pleural effusion (pleural fluid; PF) or to adenosine (in the form of NECA), prostaglandin E2 (PGE2) or forskolin to detect the induction of CD73 expression. PF treatment
was also carried out in the presence of adenosine-receptor or
PGE2-receptor inhibitors.
Results: We found significant CD73 expression on CD14+ cells
present in pleural exudate but not in peripheral blood of patients with malignant mesothelioma. The observed co-expression of CD39 and CD73 makes these tumour-associated monocytes uniquely capable of metabolising pro-inflammatory ATP
to immunosuppressive adenosine. We also found that CD73 can
be upregulated on normal CD14+ cells upon exposure to PF in
a dose- and cell type-dependent manner, as this effect was not
observed on T cells. Cyclic-AMP-inducers, forskolin and PGE2,
as well as adenosine, were also able to induce CD73 expression. Inhibition of adenosine A2a receptor or PGE2-receptors
EP2 and EP4 abolished CD73 induction by PF on monocytes,
demonstrating a self-amplifying loop by adenosine via cAMP
signalling in mesothelioma.
Conclusion: Our findings point towards a cross-talk between
adenosine, PGE2 and cyclic AMP in the regulation of CD73
expression on tumour-associated monocytes, potentially
leading to sustained adenosine production. This may contribute
not only to enhanced tumour growth but also to the immunosuppressive nature of mesothelioma and should be considered
when designing new therapeutic approaches.
Keywords: adenosine, immune cells, immunosuppression
Methods: To construct this model we used a mesothelioma
cell line generated in house from tumour tissue (#18), primary
normal lung fibroblasts (AG-02603; which we anticipate will be
‘hijacked’ by the tumour to form tumour-promoting stroma) and
primary endothelial cells (HUVECs). These cells were cultured
in low-adherence 96 well U bottomed plates to aid spheroid
formation. We assessed cell distribution within the spheroids 5
days after initial cell seeding by paraffin embedding, sectioning
and subsequent H&E staining. Spheroid growth was determined
using a 3H-thymidine incorporation assay to measure proliferation. Cytotoxic T cell killing assay measuring 51Chromium
release was used to assess susceptibility of the spheroids to T
cell mediated killing.
Results: We have successfully generated tumour only (1
component), tumour and fibroblast (2 component) and tumour,
fibroblast and endothelial cell (3 component) spheroids. After
5 days, 2 component spheroids assume a structure that is
comparable to tumours in vivo. This includes a necrotic core,
proliferating cells to the periphery of the sphere, mucin deposits
and an outermost layer of secretory mesothelial cells. The
2 and 3 component spheroids had a growth advantage over
the 1 component spheroids. We also found the 1 component
spheroids to be sensitive to T cell killing. However, as predicted,
the 2 component spheroids are significantly more resistant to T
cell killing in comparison. Furthermore, this protection is further
enhanced by the inclusion of endothelial cells in the 3 component spheres. The mechanism of this immuno-resistance,
acquired by the tumour cells in 2 and 3 component spheroids,
is currently the subject of further study.
Conclusion: In conclusion, we have generated a 3D mesothelioma tumour model with a more accurate basis than current in
vitro tumour models. We were able to mimic the growth and
survival advantage conferred by stromal and endothelial cells
and consequent resistance to immune attack. This model can
be widely used to accelerate the screening of novel therapies
that are of potential benefit to patients.
Keywords: T cells, spheroids, immuno-resistance
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MS16.06: IMMUNE ESCAPE CORRELATES
STRONGLY WITH AN INFLAMED PHENOTYPE IN
MS16.07: EXPLOITING IMMUNE CHECKPOINT
BLOCKADE TO DEVELOP EFFECTIVE THERAPY FOR
Yi-Hung Carol Tan1, Arun Khattri2, Zhixiang Zuo2, Aliya Husain3 ,
Hedy Kindler2, Tanguy Y. Seiwert2
Scott Fisher1, Jessica Solin1, Thomas Casey1, Willem J. Lesterhuis1, Sally M. Lansley2, Bruce Robinson1, Richard Lake1
Medicine, The University of Chicago, Chicago, AL, UNITED
STATES OF AMERICA, 2Medicine, The University of Chicago, Chicago, IL, UNITED STATES OF AMERICA, 3Department of Pathology, The University of Chicago, Chicago, IL, UNITED STATES OF
AMERICA
1
1
Objectives: Malignant mesothelioma (MM) is commonly
associated with an inflammatory reaction, although the specific
patterns of immune escape remain incompletely understood.
We used emerging, high-fidelity gene expression data from the
TCGA mesothelioma cohort to interrogate subgroups based on
expression of immune related genes, and determined associated immune escape mechanisms.
Methods: RNA-seq data from 25 MM were analyzed using gene
signatures representative of T-cells, NK-cells, neutrophils, and
dendritic cells/macrophages, as well as genes associated with
immune escape (immune checkpoints and cellular immune
escape). The cellular de-convolution algorithm CiberSorter was
also applied. Using this unsupervised gene set, hierarchical
clustering was performed to identify intrinsic immune subgroups. Groups correlated with T-cell inflammation (Kindler,
ASCO 2014), based on the 12-gene inflammation signature
(Harlin/Gajewski, CR 2009).
Results: MM tumors readily clustered into two large groups:
inflamed and non-inflamed. 35% of tumors demonstrated high
levels of inflammation (group 1), presence of all four immune
cell components, and 80% of tumors had a TCIP-high phenotype. Non-inflamed tumors (group 2) showed low immune cell
related gene expression and were 85% non-T-cell inflamed.
Prominent immune escape was present in all group 1 tumors,
including expression of PD-1/PD-L1, CTLA4, LAG3, and FOXP3
(however not B7H3). Inflammation strongly correlated with
presence of immune escape (functional immune response) in
group 1 while group 2 tumors exhibited neither infiltration with
immune cells nor immune escape (immunological ignorance).
There was no correlation of PD-L1 expression with either
macrophage infiltration or degree of CD8 T cell infiltration,
suggesting primarily tumor based PD-L1 expression, which was
confirmed using multi-color immunofluorescence imaging.
Conclusion: MM can be classified into inflamed/group 1 and
non-inflamed/group 2 tumors. Group 1 tumors show simultaneous infiltration with multiple immune cell components and
prominent immune escape. Inflamed and non-inflamed tumors
may require different treatment strategies for immunotherapy.
Keywords: gene signatures, mesothelioma, RNA-seq, Immunotherapy
Perth, WA, AUSTRALIA, 2Pleural Medicine Unit, Institute for Respiratory Health, Perth, WA, AUSTRALIA
Objectives: Immune checkpoint blockade (ICPB) is effective
against melanoma and is now being applied to other cancers,
including mesothelioma. ICPB therapies work by using antibodies to block molecules that negatively regulate T cell function,
thus promoting and prolonging anti-tumour immunity. Current
ICPB therapies are mostly focused on two key checkpoint molecules; CTLA-4 and PD‑1 and dual blockade appears superior to
single agent. However, at least 22 other checkpoint molecules
have been identified and the best combinations for each cancer
have yet to be defined. Using our well established preclinical
mesothelioma models, this study aims to characterise the expression profile of a subset of checkpoint molecules on immune
cells present within mesothelioma tumours to identify the best
combination of checkpoint blockade antibodies for effective
therapy against mesothelioma.
Methods: Tumours harvested from mice inoculated with AB1
and AE17 mesotheliomas were assessed by multiparameter
flow cytometry to characterise expression of immune checkpoint molecules (CTLA-4, OX40, TIM-3, GITR and PD-1) on
immune cell subsets within the mesothelioma tumour microenvironment. Based on these data, tumour bearing mice
were treated with antibodies either alone or in combination to
identify the best therapies for anti-tumour immunity against
mesothelioma in two mesothelioma tumour models.
Results: OX40 and CTLA-4 are highly expressed on tumour resident regulatory T cells (Treg) compared to tumour infiltrating
lymphocytes (CD4 and CD8 TILs). Conversely, increased PD-1
expression was observed for non-Treg CD4 and CD8 TILs compared to Treg, while broad TIM-3 expression was observed on
both effector and regulatory T cells. We observed a significant
delay in tumour growth and associated improvement in overall
survival in mice that were treated with either αCTLA-4 or αGITR
alone, but not in mice treated with αOX40, αPD-1 or αTIM-3.
Combining αCTLA‑4 with αOX40 provided no additional benefit
in tumour growth delay over CTLA-4 alone. Conversely, a small
delay was observed when PD-1 and TIM-3 were combined,
suggesting that the dose/scheduling of this combination treatment warrants further investigation. These combinations are
currently being tested in an orthotopic (intrapleural) mesothelioma mouse model.
Conclusion: Immune checkpoint blockade is a significant
breakthrough, offering promise for the treatment of many solid
tumours including mesothelioma. Data from our pre-clinical
mouse models suggest that immune checkpoint molecules
are highly expressed on tumour infiltrating immune cells and
that immunotherapies that target these molecules can delay
mesothelioma development and improve survival. We are continuing to refine our treatment protocols to ultimately identify
which combination of immune checkpoint molecules are best
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ABSTRACT BOOK
for generating effective immunity against mesothelioma in both
subcutaneous and orthotopic models. These findings will be
fundamental to inform the rational design of future clinical trials
that utilise ICPB for the treatment of mesothelioma.
Keywords: Immunotherapy, immune checkpoint blockade,
mouse models
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POSTER SESSIONS
PP01: POSTER MIXER AND POSTER DISCUSSION SESSION 1
MONDAY, MAY 2, 2016
18:00 – 19:30
PP01.01: SYSTEMIC INFLAMMATION
CONSISTENTLY PREDICTS ADVERSE OUTCOME
BASED ON MULTIPLE BIOMARKERS IN PATIENTS
WITH MESOTHELIOMA
Michael Mcgettrick1, Selina Tsim2, Kevin Blyth2
Respiratory Department, Queen Elizabeth University Hospital,
Glasgow, Glasgow, UNITED KINGDOM, 2Respiratory Department,
Queen Elizabeth University Hospital, Glasgow, UNITED KINGDOM
1
Objectives: Predicting survival in patients with Malignant
Pleural Mesothelioma (MPM) is challenging. Inflammatory
biomarkers including Neutrophil-to-Lymphocyte ratio (NLR),
Platelet-to-Lymphocyte ratio (PLR) and the modified Glasgow
Prognostic Score (mGPS), which integrates CRP (</>10mg/l)
and Albumin (</>35g/l) into a 3-point score, have proven inconsistent prognostic tools in MPM. We compared the prognostic
values of NLR, PLR and mGPS to established tools (histology,
Performance Status (PS) and EORTC Prognostic Score (EPS)).
Methods: 213/1250 patients with archived MPM tissue were
identified retrospectively.743/1250 (diagnosed pre-2008, prior
to electronic records), 213/1250 (from a different health-board)
and 11/1250 (non-pleural MPM) were excluded. Relevant data
were collected using electronic records. NLR and PLR were dichotomised using established cut-points. Kaplan-Meier survival
methodology (log-rank or log-rank for trend) and hazard ratios
(HR) were used to assess prognostic influence and quantify
risks of death.
Results: Mean age was 73 (±11) years, 81% were male. 63%
had Epithelioid, 19% Sarcomatoid, 8% Biphasic and 10%
not-specified sub-type. PS at diagnosis was 0-1 in 43%, 2-3 in
17%, 4 in 1% and not recorded in 40%. Mean NLR, computable
in 95%, was 5.7 (±4.4). Mean PLR, computable in 95%, was
309 (±205). mGPS was 0 in 56/283 (20%), 1 in 87/283 (31%),
2 in 95/283 (33%) and not computable in 45/283 (16%). EPS
was computable in 180/283 (63%) and high risk in 50/180
(28%). The prognostic significance of NLR, PLR and mGPS are
summarised in Figure 1. Interestingly, the HR in patients with
systemic inflammation (NLR >5, PLR > 300 or mGPS 1 or 2)
relative to those without was similar (around 1.6). EPS predicted
survival with similar weight to inflammatory indices (chi2 8.038,
p=0.0046) and associated HR in high-risk patients (1.68 (1.16
– 2.3)). PS and histological subtype appeared more powerful
predictors of outcome than any inflammatory index (chi2 113.1,
p ≤0.0001, HR in PS 4 and PS 2-3, 28.3 and 1.84, relative to PS
0-1 respectively and chi2 17.75, p=0.0005, HR in Sarcomatoid
and Biphasic, 2.22 (1.51 – 3.28) and 1.65 (1.08 – 3.23), relative
to Epithelioid MPM respectively).
Conclusion: Systemic inflammation is consistently associated with survival disadvantage in MPM, when measured using
different biomarkers. Although the strength of this association
is modest it appears similar to EPS, a validated prognostic tool.
PS and histology are more strongly associated with outcome.
It is not clear from the current data whether the association
between inflammation and outcome relates to co-morbidities or
tumour biology. This merits further study.
Keywords: Biomarkers, Inflammation
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PP01.02: AN UPDATE ON DIAPHRAGM: A
PROSPECTIVE, MULTI-CENTRE STUDY OF
FIBULIN-3 AND SOMASCAN AS BIOMARKERS IN
MESOTHELIOMA
Selina Tsim1, Caroline Kelly2, Laura Alexander2, Ann Peek2, Jim
Paul2, Rosemary Woodward3 , John E. Foster3 , Kevin Blyth1
Respiratory Medicine, Queen Elizabeth University Hospital,
Glasgow, UNITED KINGDOM, 2Cruk Clinical Trials Unit, Beatson West of Scotland Cancer Centre, Glasgow, UNITED KINGDOM, 3Clinical Research Imaging Facility, Queen Elizabeth
University Hospital, Glasgow, UNITED KINGDOM
1
Objectives: Diagnostics in Malignant Pleural Mesothelioma
(MPM) remain difficult and accurate non-invasive biomarkers
to guide invasive histological sampling are required. SOMAscan
is a 13-protein serum classifier with encouraging diagnostic
performance (AUC 0.95) in a retrospective surgical series but
this has not been assessed prospectively. Fibulin-3 is measured
by ELISA in plasma and has shown similar promise (AUC 0.99),
but these results have not been replicated in subsequent retrospective series. The primary objective of DIAPHRAGM (Diagnostic and Prognostic Biomarkers in the Rational Assessment
of Mesothelioma) is to prospectively assess the diagnostic and
prognostic value of these biomarkers, relative to Mesothelin
and volumetric tumour assessment using Magnetic Resonance
Imaging.
Methods: A prospective multi-centre observational study is
currently being conducted in 20 centres across the U.K. and Ireland, all of whom have evolved pleural diagnostic services. The
primary end-point is the sensitivity and specificity of SOMAscan
and Fibulin-3 for MPM. The study is powered based on available
results in 120 patients with MPM, requiring recruitment of a
projected 600 patients with Suspected Pleural Malignancy
(SPM), assuming a 20% Mesothelioma incidence rate. This reflects the study design, which requires biomarkers to be drawn
prior to pleural biopsy, pleurodesis or other intervention that
might affect results, which also mirrors the probable timing of
sampling for a tumour marker in clinical practice. Inclusion criteria are SPM and informed written consent. Exclusion criteria
are insufficient fitness for diagnostic sampling or an inter-costal
chest drain in-situ or within the preceding 3 months. Neither
asbestos exposure nor pleural plaques are inclusion criteria,
given the significant incidence of MPM in patients without these
features. 109 asbestos-exposed controls (AEC) are also being
recruited in Glasgow and the study includes extensive sample
banking for future testing of MPM biomarkers.
Results: The first recruiting centre (Glasgow) opened to recruitment on 31st December 2013. Since then, 428 patients have
been recruited to the SPM arm (71% of original target) and 73
recruited (67% of target) to the AEC arm. An interim, blinded
statistical review indicates that the incidence rate of MPM so
far, is lower than predicted (20%), with a current rate of 13%.
Significant progress has been made with the development of
quantitative volumetric MRI methods and further validation of
Early Contrast Enhancement, a recently reported and novel
perfusion-based MRI biomarker of PM.
Biomarker results will be compared with Mesothelin, the current best performing test, and volumetric assessment of tumour
burden. The lower incidence of MPM recorded will require a
study extension for 12 months, allowing completion in December 2016. The banking of prospectively collected, well-phenotyped samples of serum, plasma, whole blood (and pleural fluid
in a proportion) will create a valuable resource for future MPM
biomarker validation and/or discovery.
Keywords: Biomarkers, Diagnostics
PP01.03: EXAMINATION OF THE SERUM SOLUBLE
MESOTHELIN-RELATED PEPTIDE (SMRP) LEVEL
IN PATIENTS WITH MALIGNANT PLEURAL
MESOTHELIOMA
Taiichiro Otsuki, Eisuke Shibata, Koji Mikami, Takayuki Terada,
Kozo Kuribayashi, Takashi Nakano
Respiratory Medicine, Hyogo College of Medicine, Nishinomiya
Hyogo, JAPAN
and aggressive malignancy of the mesothelium. MPM is difficult
to detect in the early stages, as most cases are already at an
advanced stage on diagnosis. This study aimed to evaluate
serum soluble mesothelin-related peptide (SMRP) levels as a
diagnostic marker for MPM .
Methods: Serum SMRP levels were measured in 99 patients
with MPM at the Hyogo College of Medicine between April 2013
and December 2015.
Results: A total of 84 men and 15 women were included in the
study. The histologic types (n) were: epithelioid (75), sarcomatoid (9), biphasic (10), and desmoplastic (5). Regarding clinical
stage, 17 patients had Stage I disease, 28 had Stage II, 22
had Stage III, and 32 had Stage IV. The median serum SMRP
level was 2.2 nmol/L (range, 0.2–81.4 nmol/L). There were 63
positive patients (64%) when we set a reference value of 1.5
nmol/L. Based on histologic type, the median SMRP levels for
epithelioid, sarcomatoid, biphasic, and desmoplastic MPM were
2.6, 3.2, 0.95, and 0.9 nmol/L, respectively. Patients with epithelioid MPM had significantly higher SMRP levels than patients
with non-epithelioid MPM (P=0.02). However, the patient with
the higher levels had sarcomatoid mesothelioma.
Conclusion: We examined serum SMRP levels in malignant
pleural mesothelioma patients. In our analysis, we found that
serum SMRP levels is a useful marker for the diagnosis of MPM.
Conclusion: DIAPHRAGM has been designed to rigorously
assess the diagnostic performance of SOMAscan and Fibulin-3
in patients presenting with suspected PM who could have MPM.
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PP01.04: PI3-KINASE PATHWAY AND MET
INHIBITION IS EFFICACIOUS IN MALIGNANT
Rajani P. Kanteti, Jacob J. Riehm, Immanuel Dhanasingh,
Frances E. Lennon, Hedy Kindler, Ravi Salgia
Medicine, University of Chicago, Chicago, UNITED STATES OF
AMERICA
Objectives: Malignant pleural mesothelioma (MPM), an aggressive cancer commonly associated with prior asbestos exposure has poor prognosis. Receptor tyrosine kinases (RTKs) such
as MET and its downstream target PI3K are overexpressed and
activated in a majority of MPMs. Here, we studied the combinatorial therapeutic efficacy of the MET/ALK inhibitor crizotinib,
with either a pan-class I PI3K inhibitor, BKM120, or with a PI3K/
mTOR dual inhibitor, GDC-0980, in MPM.
Methods: The MPM cells were treated with crizotinib, BKM120,
or GDC-0980 alone or in combination. Relative levels of downstream signaling molecules were determined by immunoblotting and cell viability by Alamar Blue assay. The mechanism of
inhibition was further studied using apoptosis assays and cell
cycle analysis. Cell motility was studied using Boydon chamber
migration assays. The effect of crizotinib, BKM120 and their
combination on in vivo tumor growth was determined using a
PDX mesothelioma model.
Results: Cell viability results demonstrated that both pleural
and peritoneal mesothelioma cell lines were highly susceptible
to the inhibitory effects of crizotinib and BKM120 when used
individually. Furthermore, the combination of crizotinib with
either BKM120 or GDC-0980 was synergistic in suppressing
growth of pleural mesothelioma cells (CI<1). Migration assays
demonstrated that treatment of MPM cells with crizotinib,
BKM120, or GDC-0980 significantly decreased MPM cell migration, while the combination of crizotinib with either BKM120 or
GDC-0980 was synergistic. In addition, treatment of MPM cells
with BKM120 alone or in combination with crizotinib induced
G2/M arrest and apoptosis. The degree of apoptosis in MPM
cells was much greater with BKM120 treatment compared to
crizotinib and their combination elicited the maximum effect.
Both crizotinib and BKM120 strongly inhibited the activity of
MET and PI3K as evidenced by the decreased phosphorylation
of MET, AKT, and ribosomal S6 kinase. In vivo studies using a
PDX mouse model showed that crizotinib and BKM120 together
acted synergistically in inhibiting tumor growth.
PP01.05: EARLY DIAGNOSIS BY CYTOLOGY
IMPROVES SURVIVAL
Sulaf A. Own1, Gunnar Hillerdal2, Katalin Dobra1, Anders
Hjerpe1
Dept Of Laboratory Medicine / Pathology, Karolinska University
Hospital, Huddinge, SWEDEN, 2Dept Of Pulmonary Diseases,
Karolinska University Hospital, Solna, SWEDEN
1
Objectives: A conclusive diagnosis of MM can be based on
effusion cytology, using the guidelines now approved by IMIG
[1]. This makes an earlier diagnosis possible, which in turn may
influence the effect of chemotherapy [2].
Methods: During 2008-2013 a total of 91 patients were
diagnosed with MM at the Karolinska University Hospital in
Stockholm. The first diagnosis was obtained by histology in 48
cases and by cytology in 43 cases. In 14 of the latter cases a
subsequent biopsy was obtained, verifying the diagnosis. All
diagnoses were supported by clinical findings, including CT
scans. Cases with sarcomatoid MM, secondary primaries and
a case lost to further treatment were excluded, studying the
importance of time for diagnosis in 77 cases of epithelioid and
mixed type MMs. Clinical data, including evaluation of responses, were retrieved from hospital archives and survival data were
obtained from the Swedish population data base.
Results: Results The median time for diagnosis was 1 month
less for cytology compared to histology. Chemotherapy was
given somewhat more often following a histological diagnosis.
Still the overall survival and the proportion of patients surviving
3 years was significantly better following a diagnosis based on
effusion cytology; among treated patients 9/26 (38%) versus
only 1/23 (4%), the median survival being 23 months and 14
months, respectively. The rate of initial responses to chemotherapy (stable disease + partial response) was slightly better in
the cytology group
Conclusion: Both our in vitro and in vivo findings suggest that
dual inhibition of PI3K and MET pathway may be a much more
effective strategy in treating MPM as compared to a single
agent.
Keywords: Mesothelioma, MET, PI3K, Crizotinib
Fig. Survival after chemotherapy of MM diagnosed by cytopath-
logy vs histopathology
Conclusion: The earlier MM diagnosis obtained with effusion
cytology improves the overall survival after chemotherapy. With
fewer rotal numbers of malignant cells, the risk that resistance
to therapy develops with time might then be less [2]. Our
findings show the importance of the cytological diagnosis and
encourage the initiation of treatment as soon as the diagnosis
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ABSTRACT BOOK
is obtained. References 1. Hjerpe A, Ascoli V, et al: Guidelines
for Cytopathologic Diagnosis of Epithelioid and Mixed Type
Malignant Mesothelioma. Complementary Statement from the
International Mesothelioma Interest Group, also endorsed by
the International Academy of Cytology and the Papanicolaou
Society of Cytopathology. Acta Cytol. 2015;59(1):2-16. Also
published in: Diagn Cytopathol. 2015;43(7):563-76, Cytopathology. 2015;26(3):142-56, and CytoJournal, 2015;12(1): 26-40. 2.
DeVita Jr. The James Ewing lecture. The relationship between
tumor mass and resistance to chemotherapy. Implications for
surgical adjuvant treatment of cancer. Cancer 1983;51:12091220
Keywords: prognosis, Early diagnosis, Effusion cytology
PP01.06: LOW MERLIN EXPRESSION AND HIGH
SURVIVIN STAINING INDEX ARE INDICATORS
FOR POOR PROGNOSIS OF MALIGNANT
MESOTHELIOMA PATIENTS
Mayura Meerang1, Karima Bérard1, Martina Friess1, Byron
K.Y. Bitanihirwe1, Alex Soltermann2, Bart Vrugt2, Emanuela
Felley-Bosco3 , Raphael Bueno4 , William Richards5, Burkhardt
Seifert6 , Rolf Stahel7, Walter Weder1, Isabelle Opitz1
SWITZERLAND, 2Institute of Surgical Pathology, University
Hospital Zurich, Zurich, SWITZERLAND, 3Department of Molecular Oncology, University Hospital Zurich, Zurich, SWITZERLAND, 4Brigham and Women’s Hospital, Boston, MA, UNITED
STATES OF AMERICA, 5Brigham and Womens Hospital, Boston,
MA, UNITED STATES OF AMERICA, 6Department of Biostatistics,
Epidemiology, Biostatistics, And Prevention Institute (ebpi),
University of Zurich, Zurich, SWITZERLAND, 7Clinic For Oncology,
University Hospital Zurich, Zurich, SWITZERLAND
1
Objectives: Alterations of the tumor suppressor Neurofibromatosis type II (NF2) have been reported in about 40% of
Malignant pleural mesothelioma (MPM) patients. NF2 (Merlin)
deficiency leads to alterations of the Hippo pathway; resulting in
activation of the oncogenic Yes Associated Protein-1 (YAP1). Our
aim was to investigate the association between these alterations and clinical outcomes.
Results: Kaplan-Meier survival curves revealed a significant
association between low cytoplasmic Merlin expression in
pre-induction CTX tissues of cohort 1 with shorter PFS and
OS (figure 1A, 1B). The same tendency was observed in the
chemotherapy naïve tissues obtained during EPP of cohort 2.
Low nuclear Merlin expression in post-CTX tissues (available
from cohort 1 only) was associated with short PFS (p=0.04) and
OS (p=0.05). High nuclear Survivin labeling indices in both preand post-CTX tissues of cohort 1 was associated with shorter
PFS (figure 1C, 1D). In cohort 2, this was associated with both
PFS and OS (p=0.046 and p=0.002, respectively). In multivariate analysis, low expression of cytoplasmic Merlin remained an
independent prognosticator for short PFS of cohort 1 [HR (95%
CI): 0.5 (0.3-0.9); p=0.001] and OS [HR (95% CI): 0.5 (0.3-1);
p=0.04]. High Survivin labeling index was an independent prognostic factor for short PFS in patients from cohort 1 [HR (95%
CI): 3.4 (1.7-6.8); p=0.006] and short OS in patients from cohort
2 [HR (95% CI): 2.35 (1.27-4.33); p=0.006].
(See next page.)
Methods: Tissue microarrays comprised of MPM tumors
derived from 2 independent MPM cohorts were employed for
this study. Immunohistochemical expression of Merlin, YAP1
and its target genes, Survivin and connective tissue growth
factor (CTGF) were assessed in both nuclear and cytoplasmic
fractions. Cohort 1 was comprised of 145 patients intended to
be treated with chemotherapy (CTX) followed by extrapleural
pneumonectomy (EPP), thus both pre- and post-CTX tissues
were available. Cohort 2 was comprised of 59 patients treated
with EPP followed by intraoperative hyperthermic cisplatin and/
or adjuvant CTX and/or radiotherapy. Marker expression was
quantified by means of labeling index (%) for nuclear Survivin
and by H-score for the other markers. The dichotomized marker
expression was tested for the association with overall survival
(OS) and progression free survival (PFS).
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Conclusion: Our findings uncover the significance of Merlin
protein expression and Survivin labeling index as prognosticators for poor clinical outcome in two independent MPM cohorts.
If confirmed, these markers may be used to identify subgroups
of patients benefitting from additional treatment.
Keywords: Survivin, NF2, Merlin, Hippo pathway, Prognostic
biomarkers
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PP01.07: MICRORNAS IN BLOOD AS BIOMARKER
OF PLEURAL MALIGNANT MESOTHELIOMA
Valentina Bollati1, Tommaso Cavalleri2, Chiara Favero1, Laura
Dioni1, Carolina Mensi3 , Claudia Bareggi4 , Lorenzo Bordini3 ,
Alessandro Palleschi5, Aldo Todaro3 , Dario Consonni3 , Angela
C. Pesatori1
Epiget, Department of Clinical Sciences And Community Health,
Università degli Studi di Milano, Milan, ITALY, 2Laboratory of
Molecular Gastroenterology, Humanitas Clinical and Research,
Rozzano, ITALY, 3Department of Preventive Medicine, Fondazione IRCCS Ca’ Granda - Ospedale Maggiore Policlinico, Milan,
ITALY, 4Unit Of Medical Oncology, Fondazione IRCCS Ca’ Granda
- Ospedale Maggiore Policlinico, Milan, ITALY, 5Thoracic Surgery
Unit, Fondazione IRCCS Ca’ Granda - Ospedale Maggiore Policlinico, Milan, ITALY
1
Objectives: Malignant Pleural Mesothelioma (MPM) is an aggressive cancer refractory to current therapies caused almost
exclusively by asbestos. New specific diagnostic markers for
early MPM diagnosis are needed. MiRNAs are single stranded
noncoding that post-transcriptionally regulate gene expression
by triggering mRNA cleavage or repressing translation. Changes
in the expression of miRNAs have been implicated in several
diseases and cancers, including MPM. miRNAs are stable
molecules that can be easily investigated in different specimens
(e.g. blood), and used as a disease biomarker. Our aim was to
determine if a specific miRNA signature in plasma may help to
discriminate between malignant pleural mesothelioma patients
(MPM) and healthy subjects with a Past Asbestos Exposure
(PAE).
Methods: We investigated a group of 23 MPM patients and
19 healthy subjects with Past Asbestos Exposure (PAE). In
this study population we screened 754 miRNAs in blood by
TaqMan™ OpenArray® Human MiRNA Panel. Than we selected
for validation, in the same groups of subjects, the top-23 differential miRNAs. RNU48 was used as endogenous control. We
used multiple linear and logistic regression models adjusted for
age, sex, BMI, and smoking habits to compare miRNAs profiling
between MPM and PAE subjects.
Results: After miRNA screening, we identified 29 differential
miRNAs in plasma. Among the top 23 differential miRNAs, 19
were validated by Real time PCR and were able to discriminate
between MPM and PEA subjects. In receiver operating characteristic (ROC) curve analysis, the three best miRNAs were
miR-103 (area under curve, AUC=0.82), miR-98 (AUC=0.82) and
miR-744 (AUC=0.81).
Conclusion: The identified signature was useful for MPM diagnosis. Further studies are needed to verify if they can be of help
for early MPM diagnosis and/or to detect high risk groups.
Keywords: microRNA, epigenetics, mesothelioma diagnosis,
Asbestos exposure
PP01.08: SEPTIN-7, LIPOMA PREFERRED
PARTNER AND TRANSALDOLASE DISCRIMINATE
NEOPLASTIC FROM PRENEOPLASTIC
MESOTHELIAL CELLS
Daniel L. Pouliquen1, Alice Boissard2, Béatrice NawrockiRaby3 , Joëlle Nader1, Philippe Birembaut3 , Marc Grégoire1,
Olivier Coqueret2, Catherine Guette2
UMR 892 INSERM / 6299 CNRS, Nantes, FRANCE, 2UMR 892
INSERM / 6299 CNRS, Angers, FRANCE, 3UMRS-903 INSERM,
Reims, FRANCE
1
Objectives: Quantitative proteomic analyses, which allow
to distinguish invasive and less aggressive stages of some
cancers, has helped to identify new diagnostic or pronostic biomarkers. To date, applied to mesothelioma, this approach has
poorly been investigated. In this study we used a biocollection
of preneoplastic and neoplastic mesothelial cell lines established from F344 rats induced with crocidolite to determine
proteomic signatures specific of each stage.
Methods: The cell lines used in this study belong to a biocollection established in 2011 from a group of F344 rats, after 136
to 415 days of induction with crocidolite administered intraperitoneally. Nine cell lines, including 4 neoplastic cell lines
(which produce tumors after orthotopic injection to syngeneic
rats), and 5 preneoplastic cell lines (not producing tumors)
representing different stages of the epithelial-to-mesenchymal
transition (EMT) according to previous transcriptomic analysis
of their expression profile, were selected for the study. Proteomic analyses were conducted using a LC-MS/MS approach with
iTRAQ labeling on 4 x 106 cells of each cell line, in comparison with a reference cell line from the biocollection showing
a typical epithelioid cobblestone morphology comparable to
normal mesothelial cells and presenting the highest expression
of Cdh1 and the lowest expression of the Acta2 and Tgfb1 genes.
The in vitro invasive properties of the four neoplastic cell lines
were also assessed using a modified Boyden chamber assay, in
comparison with preneoplastic cells.
Results: Among the 950 detected, 57 proteins of interest were
identified, characterized by a significant increase or decrease
relative to the reference cell line. Three of the 4 neoplastic cell
lines, characterized by their invasive properties both in vitro in
the Boyden chamber assay, and in vivo after transplantation to
syngeneic rats, showed significant higher levels of transaldolase
(taldo1). Conversely, all preneoplastic cell lines and the fourth,
non-invasive, neoplastic cell line, M5-T2, were characterized by
a significant higher level of septin-7 (sept-07) and lipoma-preferred partner homolog (Lpp). The peculiar case of M5-T2 was
also associated with the highest level of Lpp. Finally, among the
three invasive neoplastic cell lines, the maximum decrease in
the content of Lpp and sept-07, and the highest level of taldo 1,
was observed for the M5-T1 cell line, which showed the highest
capacity of migration in the Boyden chamber and the highest
metastatic potential in vivo.
Conclusion: These preliminary results obtained on 9 cell lines
open up interesting prospects for complementary studies on
the whole biocollection, with the aim to characterize the proteomic signature of the different stages of mesothelial carcinogenesis.
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Keywords: Preneoplastic, Neoplastic, Quantitative proteomic
analyses, Cell lines
PP01.10: PROGNOSTIC SIGNIFICANCE OF
MITOTIC ARREST DEFICIENT 2-LIKE PROTEIN 1 IN
Aaron S. Mansfield1, Tobias Peikert2, Justin Moser3 , Anja
Roden4
PP01.09: COL3A1, A NEW POTENTIAL
IMMUNOHISTOLOGICAL MARKER FOR
THE DIAGNOSIS OF MALIGNANT PLEURAL
MESOTHELIOMA
Charly Liddell1, Fabien Gueugnon2, Jean-Michel Nguyen2,
Christine Sagan1, Laurent Cellerin3 , Marc Grégoire2, Christophe
Blanquart 2
Pathology, CHU de Nantes, Nantes, FRANCE, 2INSERM UMR 892
/ CNRS 6299, Nantes, FRANCE, 3Pneumology, CHU de Nantes,
Nantes, FRANCE
1
and aggressive cancer related to asbestos exposure. The diagnosis is performed by immunohistology. However, in some cases the differential diagnosis of MPM from lung adenocarcinoma
(ADCA) remains difficult. Using a transcriptomic approach, we
identified Col3A1 as a new potential biomarker for MPM. The
aim of this study was to validate Col3A1 for the differential
diagnosis of MPM from lung ADCA by immunohistology
Methods: We first studied the higher mRNA expression of
Col3A1 in MPM compared with lung ADCA using our biocollection of cell lines established from pleural fluids of patients.
We performed immunofluorescence experiments to confirm
the mRNA results. Then, Col3A1 labeling was performed on 14
MPM and 15 lung ADCA tumor slides to evaluate the diagnostic
value of this biomarker.
Results: Col3A1 mRNA expression was demonstrated to be
specific for cells of mesothelial origin. We showed that the
mRNA expression of Col3A1 is as effective as that of WT1 (error
rate (ER): 0.0526) and better than calretinin (ER: 0.263), podoplanin (ER: 0.2105) and keratin 5 (ER: 0.3157) in differentiating
MPM cells from lung ADCA cells. The diagnostic potential of
Col3A1 to differentiate MPM from lung ADCA was confirmed by
immunohistology, with a specificity of 92.85% and a sensitivity
of 93.30% for the percentage of stained cells.
Conclusion: This study validates Col3A1 as a new potential
MPM biomarker for immunohistology. Thus, this biomarker
could be included in the panel of biomarkers currently used for
MPM diagnosis after independent validation.
Keywords: diagnosis, mesothelioma, biomarker, immunohistology
Medical Oncology, Mayo Clinic, Rochester, MN, UNITED STATES
OF AMERICA, 2Pulmonary And Critical Care Medicine, Mayo
Clinic, Rochester, MN, UNITED STATES OF AMERICA, 3Internal
Medicine, Mayo Clinic, Rochester, MN, UNITED STATES OF AMERICA, 4Laboratory Medicine And Pathology, Mayo Clinic, Rochester, MN, UNITED STATES OF AMERICA
1
Objectives: Mitotic Arrest Deficient 2-Like Protein 1 (MAD2L1) is
a critical component of the spindle assembly checkpoint, which
delays mitosis until all kinetochores are attached to a mitotic
spindle ensuring appropriate segregation of sister chromatids.
Overexpression of MAD2L1 results in stabilization of Securin
and Cyclin B, delays exit from mitosis and promotes aneuploidy
through nondisjunction events. MAD2L1 mutations are rare, but
overexpression of MAD2L1 is common in human malignancies.
Recently, MAD2L1 expression has been identified as a potential therapeutic target in malignant mesothelioma (MM) and
a prognostic marker in early stage lung cancer. We sought to
characterize the expression of MAD2L1 in MM and to determine
its prognostic significance.
Methods: We developed a tissue microarray (TMA) with 160
cases of MM with 0.6mm cores in triplicate. Immunostaining
was performed at Mayo Clinic’s Pathology Research Core using
the Leica Bond RX stainer. The MAD2L1 antibody (Bethyl Laboratories IHC-00412 rabbit, polyclonal) was diluted 1:75 in Background Reducing Diluent (Dako) for 30 minutes. The detection
system used for MAD2L1 was the Envision Flex System (Dako).
Percent expression was scored as 0 (none), 1 (1-25% tumor
cell expression), 2 (26-50%), 3 (51-75%) or 4 (76-100%); the
intensity was scored as 0 (negative), 1 (weak), 2 (moderate) or 3
(strong). The average H-score (percent expression score multiplied by intensity score) for the three cores was determined for
each case. Survival was estimated with Kaplan-Meier curves by
quartiles and compared with a log-rank test. All but 25 subjects
were followed until death.
Results: 109 males (68%) and 51 females (32%) had a median
age of 66 years (interquartile range, 58-73). The cases were
epithelioid (n=99, 62%), biphasic (n=26, 16%), sarcomatoid
(n=25, 16%) or not further specified (n=10, 6%). A broad
range of MAD2L1 expression was noted (median H-score=8,
interquartile range 5-9, range 0-12). Kaplan-Meier estimates
demonstrated similar survival of cases with the three highest
quartiles of MAD2L1 expression. Therefore, these cases were
grouped together and their survival was compared to that of
cases in the lower quartile. The survival of patients with the lowest quartile of MAD2L1 expression (n=39, median=13 months,
mean=37 months) was significantly greater than that of patients
with the three highest quartiles of MAD2L1 expression (n=121,
median=10 months, mean=17 months; p=0.05); with most of
the difference seen in the tails of the curves (Figure 1).
Conclusion: Increased MAD2L1 expression is a negative prognostic marker in MM. The therapeutic modulation of MAD2L1 is
being explored.
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significantly higher in the BPD group than in MPD or MPM
patients (p<0.0001). At the opposite, pleural BDNF were significantly higher in MPM patients vs. MPD patients or BPD patients
(p=0.0002).
Conclusion: Plasma fibulin-3 and BDNF seemed of little interest for MPM diagnosis, showing some discrepancies with previous published results for fibulin-3. Pleural fibulin-3 level might
have a prognostic value in pleural diseases. Increased levels of
BDNF in pleural effusions are associated with malignancy and
more particularly with MPM for highest BDNF levels.
Keywords: biomarker, BDNF, fibulin-3, mesothelioma
Keywords: prognosis, mesothelioma, mad2l1
PP01.12: TARGET TRIAL
Duneesha De Fonseka1, Louise Allen1, Nick Maskell2
PP01.11: BRAIN-DERIVED NEUROTROPHIC
FACTOR (BDNF) AND FIBULIN-3 AS BIOMARKERS
FOR DIAGNOSIS OF MALIGNANT PLEURAL
MESOTHELIOMA (MPM)
Sarah Benziane1, Patrick Smeele2, Sophie Deshayes3 ,
Anne-Laure Chéné4 , Camille Munck1, Marie C. Willemin1, Eric
Wasielewski1, Arnaud Scherpereel1, Marc Grégoire2, Christophe
Blanquart2
Pulmonary and Thoracic Oncology Department, Hospital of the
University (CHU) of Lille, Lille cedex, FRANCE, 2Centre de Recherche contre le Cancer Nantes et Angers, University of Nantes,
CNRS UMR 6299, Inserm U892, Nantes cedex, FRANCE, 3Centre
de Recherche contre le Cancer Nantes et Angers, University of Nantes, CNRS UMR 6299, Inserm U892, Nantes cedex ,
FRANCE, 4Digestive and Thoracic Oncology Department, Laënnec
Hospital, CHU of Nantes, Nantes, FRANCE
1
Objectives: MPM is a rare tumor with poor prognosis, usually
associated with previous asbestos exposure. Its diagnosis,
usually based on histology, obtained by invasive procedures,
may be tricky. To date, no diagnostic biomarker was validated
in routine. In this study, we aimed at evaluating two potential
biomarkers, BDNF and fibulin-3.
Methods: observational, retrospective study in 2 French centers. Plasma and pleural effusion samples were collected from
3 different groups : patients with « benign pleural diseases »
(BPD), patients with malignant pleural diseases excluding MPM
(MPD), or MPM patients. Biomarker levels were determined
using ELISA assays (Human BDNF DuoSet (R&D Systems) and
FBLN3 (USCN)).
Results: 310 patients (76 BPD, 108 MPD, 126 MPM) were
recruited. Plasma fibulin-3 levels were significantly higher in
the BPD group than in MPD or MPM patients (p<0.0001), and
in MPM vs. MPD patients (p=0.0056). There was no difference
between the 3 groups for pleural fibulin-3 levels. However,
patients with highest pleural fibulin-3 levels at the time of
diagnostic tend to have a shorter survival (p=0.06), suggesting
a possible prognostic value. Plasma BDNF levels were also
Academic Respiratory Unit, North Bristol NHS Trust, Bristol,
UNITED KINGDOM, 2Academic Respiratory Unit, University of
Bristol, Bristol, UNITED KINGDOM
1
Objectives: TARGET trial is a randomised controlled trial designed to compare the diagnostic yield of PET-CT guided pleural
biopsy versus CT-guided pleural biopsy in suspected pleural
malignancy, where patients have already had one non-diagnostic biopsy. Diagnosis of pleural malignancy, particualrly
mesothelioma can be challenging in the absence of pleural
fluid to perform a thoracoscopy for direct visualisation and
biopsy of the pleura. Computed Tomography (CT) or Ultrasound
(US) guided biopsy of the pleura are two of the commonest
techniques used in this situation but the diagnostic rate is low.
Diagnostic imaging in pleural malignancy remains a significant
challenge. PET-CT scanning has proven itself a useful tool in
diagnosing and staging lung cancer. It identifies areas of high
metabolic activity, which is a feature of malignant disease, by
highlighting areas of uptake of the radio labelled glucose analogue Fluorodeoxyglucose (FDG). We hypothesise that targeting
the CT guided biopsy to these highlighted areas on PET-CT may
increase the diagnostic yield of pleural biopsies.
Methods: This multi-centre randomised controlled study
conducted in the UK will recruit patients from 10 respiratory
departments over a 24 month period or until 78 patients have
been recruited. Patients will be randomised either to receive a
PET targeted biopsy or a standard CT guided biopsy using an
online randomisation tool. The experimental group will undergo
a PET scan which will be reviewed by a local radiologist to
identify the most suitable area for biopsy, then a CT guided
biopsy targeted to the afore highlighted area. The standard CT
group will have a repeat CT and a biopsy from a site identified
as per the local radiologist. Standard Operating Procedures
(SOP) will be in place to minimise variation. All patients will be
followed-up for a 12 month period from randomisation. The
study is NIHR (Research for Patient Benefit) funded.
Results: The trial opened to recrutiment in September at North
Bristol NHS Trust and is currently in the set-up phase of 5 other
UK trusts. Results of this study will be due to in 2019.
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Conclusion: If this superiority study is proven successful,
it would shorten the patient’s cancer journey and reduce
the number of invasive investigations they undergo. An early
diagnosis may allow more patients to have life prolonging
chemotherapy that they may not be able to have at a later stage
in disease if they are too unwell. Mesothelioma patients have a
very poor survival and often some of this time may be spent in
hospital. An early diagnosis would allow patients more time to
enjoy the financial benefits of the compensations they receive
following a diagnosis of mesothelioma.
Keywords: pleural mesothelioma, PET-CT scan, CT guided
biopsy
Fig. Effects of Carboplatin (C), Pemetrexed (P) on a primary
isolate of malignant mesothelioma. In this case both drugs
cause arrest in early S-phase when compared to the untreated
control. This effect is masked when the two drugs are combined, instead resulting in a broader G0/1 peak.
PP01.13: EX VIVO EFFECTS OF PEMETREXED AND
CARBOPLATIN ON MALIGNANT MESOTHELIOMA
CELLS
Carl-Olof Hillerdal1, Rita Ötvöss1, Tände Szatmari1, Katalin
Dobra1, Anders Hjerpe2
Dept Of Laboratory Medicine / Pathology, Karolinska Institutet,
Huddinge, SWEDEN, 2Dept Of Laboratory Medicine / Pathology,
Karolinska University Hospital, Huddinge, SWEDEN
1
Objectives: The combination pemetrexed and carboplatin is
the standard treatment for malignant mesothelioma worldwide.
Malignant mesotheliomas have an objective response rate of
40% to this combination. The ex vivo analysis of the sensitivity
to these drugs could be helpful in predicting the effects of individual tumours, and thereby provide a basis for future personalized choice of therapy. However, when testing mesothelioma
cell lines, 48 hours’ drug exposure to pemetrexed had no effect
when evaluating with a live/dead assay based on remaining
mitochondrial activity, while the S-phase resulting arrest could
be traced by cell cycle analysis.
Methods: Tumor cells were isolated from pleural effusions
obtained from patients with malignant mesothelioma and
grown as short-term cultures in IMDM medium, supplemented
with 20% FCS. These cultures were then incubated for 48 or
72 hours, with pemetrexed, carboplatin and their combinations
added to the culture medium. The effects of these exposures
were analysed with a cell cycle assay based on flow cytometry.
For comparison the development of apoptosis was monitored
with a flow cytometer annexinV/PI assay and cell survival with a
mitochondrial activity assay.
Results: Exposure of mesothelioma cell isolates to pemetrexed
during 48 hours regularly caused an increased proportion of
cells in early s-phase. A similar effect could in some isolates
also be obtained after incubation with carboplatin alone. While
in some cases carboplatin caused an increased proportion of
apoptotic cells and a corresponding decrease in cell survival,
no such effect could be seen after incubation with pemetrexed,
even when the exposure time was prolonged to 72 hours.
Interestingly, when the two dugs were combined, the effects
were masked, probably due to confluence of G0/1 and S-phase
peaks. Conclusion: All cytotoxic drugs have a unique modus operandi. At difference with many other cytotoxic drugs the relatively
short 48 hours’ incubation in medium containing pemetrexed
will not result in apoptosis and decreased cell survival. The
estimation of sensitivity to this drug ex vivo is best performed as
a cell cycle analysis, without simultaneous exposure to carboplatin. We now correlate this effect on S-phase progression with
the actual responses to given therapy.
Keywords: chemotherapy, Ex vivo analysis, prediction, mesothelioma
PP01.14: DIAGNOSING MESOTHELIOMA VIA
CHEST WALL MOTION ANALYSIS TECHNOLOGY
Ghazi Elshafie1, Prem Kumar2, Andrea Aliverti3 , Madava
Djearaman1, Babu Naidu1
Heartlands Hospital, Birmingham, UNITED KINGDOM, 2University of Birmingham, Birmingham, UNITED KINGDOM, 3Politecnico
di Milano, Milano, ITALY
1
Objectives: Mesothelioma carries a poor prognosis. . Differentiating benign from malignant pleural disease can be challenging, and may require invasive surgery. We aim to evaluate
the effects of benign and malignant pleural disease on chest
wall motion using OEP, this may constitute a useful tool in the
diagnostic pathway.
Methods: Optoelectronic plethysmography (OEP) was used to
measure chest wall motion of 16 patients recruited to this study.
The measurements were performed before any pathological
diagnosis. Pathological diagnosis was obtained via surgical
pleural biopsy. After the pathological diagnosis, they were
divided into 3 groups. A pleural disease-free (n=4), an empyema
group (n= 6) and a mesothelioma group (n=4). A radiological
assessment was performed in all patients.
Results: The relative contribution of the diseased part of the
pulmonary ribcage motion to the overall pulmonary ribcage
motion was significantly lower in the mesothelioma group, 19
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+/- 17 % compared the control and empyema group 50 +/- 3 %
and 44 +/- 7 % (p 0.01 and p 0.01) respectively (figure 1). The
radiological diagnosis matched the pathological diagnosis in
only 30% of all patients. When the diseased side of the chest
contributed ≤ 36% of the overall chest wall motion all patients
had a pathological diagnosis of mesothelioma.
Conclusion: This is the first data to describe the effect of
empyema and malignant mesothelioma on chest wall mechanics, and proved that OEP, is more predictive than conventional
radiology, and could be used in clinical practice to diagnose
those diseases.
analyzed by C4d immunohistochemistry. We compared the
correlation of C4d levels with the clinicopathological characteristics of the MPM patients.
Results: We found no evidence of C4d labeling on tumor cells
in the MPM tissue specimens. However, germinal centers of
lymphoid structures within the tumor stroma showed C4d positivity. Circulating levels of C4d were not significantly increased
in MPM patients when compared to healthy controls or patients
with non-malignant pleural diseases. Importantly, after dividing
the patient cohort by using the cutoff of 1.5µg/mL, patients
with low C4d levels at diagnosis showed a very strong tendency
towards longer overall survival. C4d levels following induction
chemotherapy were remarkably higher in patients with stable
disease or progressive disease (SD/PD) when compared to
patients with minor or major positive chemotherapeutic response (MR) (SD/PD: 1.94±0.34 µg/mL, MR: 0.70±0.51 µg/mL;
p=0.026). Interestingly, several inflammation-related prognostic markers including fibrinogen, CRP and albumin showed no
significant correlation with chemotherapeutic response.
Conclusion: Circulating plasma level of C4d is a potential
new prognostic biomarker in MPM patients. Since prediction
of response after induction chemotherapy in MPM patients is a
challenging task, our data suggest that measuring circulating
C4d levels may help to risk-stratify patients following induction
chemotherapy.
Keywords: complement activation, response evaluation, prognostic biomarker
Keywords: Chest Wall, mesothelioma, Optoelectronic plethysmography
PP01.15: CIRCULATING LEVEL OF THE
COMPLEMENT COMPONENT 4D (C4D)
CORRELATES WITH CHEMOTHERAPEUTIC
RESPONSE AND SURVIVAL IN MPM PATIENTS
Thomas Klikovits1, Paul Stockhammer1, Julia Kodnar2, Viktoria
Laszlo1, Yawen Dong1, Mir A. Hoda1, Walter Klepetko1, Balazs
Dome1, Rudolf Oehler2, Balazs Hegedus1
PP01.16: VOC ANALYSIS IN HEADSPACE AIR OF
MESOTHELIOMA AND LUNG CANCER CELL LINES:
A COMPARATIVE LITERATURE STUDY
Sabrina Lagniau1, Kevin Lamote2, Karim Y. Vermaelen2, Jan P.
Van Meerbeeck3
Internal Medicine, Ghent University, Ghent, BELGIUM, 2Respiratory Medicine, Ghent University Hospital, Ghent, BELGIUM, 3Thoracic Oncology, Antwerp University Hospital, Edegem, BELGIUM
1
Objectives: Circulating levels of the degradation product of
C4 (C4d) were found to be prognostic in lung adenocarcinoma
patients. Since there is limited information available about the
role of complement activation in the progression of MPM we
analyzed the circulating and tissue levels of C4d in a cohort of
MPM patients.
Objectives: Early detection of malignant pleural mesothelioma
(MPM) and lung cancer is important in order to improve the disease’s management. Research has focused on volatile organic
compounds (VOCs) in breath to serve as early screening tools.
Although several VOCs have been identified, it is not yet clear
which VOCs arise from the neoplastic cells themselves or from
the host response. The analysis of headspace air of mesothelioma and lung cancer cell lines can therefore be useful. The
goal of this study was to perform a literature search in order
to compare different methods for headspace analysis and to
identify tumour-specific VOCs which could serve as interesting
biomarkers.
Methods: Plasma samples from MPM patients (n = 56) were
measured by ELISA for C4d levels. Additionally, healthy volunteers (n = 19) and patients with non-malignant pleural diseases
(n = 14) were also included. In order to investigate local C4d
expression, FFPE tissue specimens from 38 MPM patients were
Methods: MEDLINE (PubMed) and Web of Science were
searched for studies concerning headspace analysis in lung
cancer and mesothelioma cell lines until January 2016. The
following keywords have been applied: “headspace analysis”
AND “lung cancer”, “headspace analysis” AND “lung cancer”
Division of Thoracic Surgery, Medical University of Vienna, Vienna, AUSTRIA, 2Anna Spiegel Center for Translational Research,
Department of Surgery, Medical University of Vienna, Vienna,
AUSTRIA
1
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ABSTRACT BOOK
AND “cell line”, “headspace analysis” AND “mesothelioma” and
“headspace analysis” AND “mesothelioma” AND “cell line”.
After removal of duplicates and manual selection of relevant
articles, 9 articles remained and are compared in Table 1.
Results: Table I: Summary. Conclusion: A plethora of VOCs is released or consumed by
cancerous cell lines, which makes these interesting to use as
biological markers in the diagnosis of lung cancer or mesothelioma. Nevertheless, no single VOC can presently be used
as stand-alone biomarker. Furthermore, the studies are not
comparable due to the use of different cell lines. This literature
study shows that in the headspace air of cancerous cell lines
the concentration of certain aldehydes (acetaldehyde), ketones (2-butanone, cyclohexanon) and alkanes is significantly
decreased or increased in comparison with the headspace of
non-cancerous cell lines or the headspace of medium. The
most frequently used method in the studied articles is GC-MS.
Because of the small number of studies and large interstudy
differences, further translational research is necessary in
order to determine which VOCs are tumour-specific, to optimize the sampling techniques and to gain information about
real tumour-specific VOCs. It will also give information about
the tumour’s metabolism. Finally, this literature study shows
that analysis of VOCs in the headspace of cancer cell lines is
promising and can be further explored for the development of
biomarkers for early disease detection.
Keywords: headspace analysis, Lung cancer, volatile organic
compounds, mesothelioma
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PP01.17: PROGNOSTIC BIOMARKERS IN A
LARGE COHORT OF PATIENTS WITH MALIGNANT
PLEURAL MESOTHELIOMA - A RETROSPECTIVE
TWO-CENTER STUDY
PP01.18: CHANGES IN MONOCYTE COUNT AND
LYMPHOCYTE-TO-MONOCYTE RATIO DURING
INDUCTION CHEMOTHERAPY CORRELATE WITH
CLINICAL OUTCOME
Thomas Klikovits1, Pietro Bertoglio2, Marcello C. Ambrogi2, Paul
Stockhammer1, Yawen Dong1, Balazs Dome1, Balazs Hegedus1,
Viktoria Laszlo1, Walter Klepetko1, Alfredo Mussi2, Mir A. Hoda1
Paul Stockhammer, Yawen Dong, Thomas Klikovits, Walter
Klepetko, Balazs Dome, Balazs Hegedus, Mir A. Hoda
Division of Thoracic Surgery, Medical University of Vienna, Vienna, AUSTRIA, 2Division of Thoracic Surgery, University Hospital of
Pisa, Pisa, ITALY
1
Objectives: The aim of this study was to identify and validate
prognostic biomarkers in a large cohort of patients with malignant pleural mesothelioma (MPM).
Methods: We performed a retrospective chart review and
included all patients with histologically confirmed MPM, treated
at two specialized centers between 1994 and 2014. The effect
of different clinical and pathological characteristics and laboratory values on patient outcome was investigated.
Results: Two-hundred ninety-one patients were identified (222
males and 69 females). Mean age at diagnosis was 64 years
(range 27-91 years). Main histological subtype was epitheloid
(n=199, 68%). Multimodality treatment, defined as macroscopic complete resection combined with chemotherapy and/or radiation therapy and/or intracavitary treatment, was performed
in 134 (46%) patients. Median overall survival in the whole
cohort was 17.7 months from diagnosis; 1-, 3- and 5-year survival rates were 65%, 28% and 19%, respectively. When using
univariate analyses, following variables were associated with a
poor prognosis: advanced age (≥70 years, p=0.001), non-epithelial histological subtype (p =0.003), fibrinogen at diagnosis
(HR 1.002, p=0.001), albumin at diagnosis (HR 0.963, p=0.001),
haemoglobin at diagnosis (HR 0.874, p=0.001), platelets at diagnosis (HR 1.003, p=0.001), leucocytes at diagnosis (HR 1.116,
p=0.001), neutrophils to lymphocytes ratio (NLR) at diagnosis
(HR 1.063, p=0.006) and platelets to lymphocyte ratio (PLR) at
diagnosis (HR 1.001, p=0.001). In the multivariate cox regression analysis, leucocyte count, fibrinogen, histological subtype
and age remained as significant co-factors.
Conclusion: Leucocyte count and fibrinogen at diagnosis were
independently prognostic in a large cohort of MPM patients.
These findings suggest that these inflammatory based prognostic markers can be utilized for improved patient selection with
regard to different therapeutic modalities.
Keywords: Biomarkers, prognostic value, therapeutic decision
Division of Thoracic Surgery, Medical University of Vienna,
Vienna, AUSTRIA
Objectives: Lymphocyte-to-monocyte ratio (LMR) and monocyte count have previously been found to be prognostic in MPM
patients. In order to validate these findings and to investigate
the clinical significance of monocyte and lymphocyte count
changes associated with induction chemotherapy, we followed
the changes in the leukocyte subpopulations during chemotherapy in a cohort of MPM patients.
Methods: Pre- and postchemotherapeutic levels of leukocytes,
lymphocytes and monocytes were collected retrospectively
from 113 patients with histologically confirmed MPM. 76 of
these patients had a positive response after chemotherapy, 37
patients had a radiologically confirmed disease progression. We
compared the data with the clinicopathological characteristics
of the MPM patients.
Results: We could confirm the prognostic relevance of LMR (LMR
≥2.74: median 50.8 months versus 17.9 months; p = 0.035) and absolute monocyte count at time of diagnosis (≥510 cells/µL: median
17.9 months versus 35.6 months; p = 0.013) in our MPM patient
cohort (n = 113) using cut-offs established in the previous studies.
In terms of chemotherapy, total leukocyte count showed a significant decrease after chemotherapy regardless to the response to
treatment (Leukocyte count - 95% CI: -0.50 to -1.92 ×109/L; p =
0.001). Relative lymphocyte count as well as relative monocyte
count showed a strong tendency for increasing after chemotherapy
(Relative lymphocyte count - 95% CI: +0.0% to +5.5%; p = 0.05;
relative monocyte count - 95% CI: 0.2% to 1.8%; p = 0.01). LMR
and absolute monocyte count did not change after chemotherapy.
When comparing patients with progressive disease and patients
with positive response to chemotherapy, we found that in each
group there was a strong tendency for a leukocyte count decrease
after induction treatment. Relative lymphocyte count as well as
absolute monocyte count did not correlate with chemotherapeutic
outcome. However, patients with a positive response after chemotherapy had a significantly reduced LMR (95% CI: -0.17 to -1.11
×109/L; p <0.01), whereas patients with progressive disease had an
elevated LMR (95% CI: -0.17 to +2.28 ×109/L; p = 0.09). Furthermore, patients with a positive response had a highly significant
increase in the relative monocyte count (95% CI: +1.2% to +2.9%;
p <0.0001) whereas patients with disease progression showed a
tendency for a decrease in their relative monocyte counts (95% CI:
+0.4% to -2.5%; p = 0.15).
Conclusion: LMR and circulating monocyte count are prognosticators in MPM patients. Furthermore, positive response
to chemotherapy is associated with a significant increase in
the relative monocyte count as well as with a LMR decrease.
Monitoring monocyte count and LMR during chemotherapy may
provide a tool to identify patients with a positive response.
Keywords: induction chemotherapy, Monocyte count, Lymphocyte-to monocyte ratio, White blood cells
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PP01.19: COMBINING CELL-FREE MICRORNAS
AND SMRP: A MULTI-BIOMARKER SIGNATURE
WITH IMPROVED DIAGNOSTIC ACCURACY
Michaela B. Kirschner1, Marissa Williams2, Sjaak Burgers3 , Mir
A. Hoda4 , Catharina M. Korse3 , Daan Van Den Broek5, Thomas
Klikovits4 , Balazs Hegedus4 , Balazs Dome4 , Michael Grusch4 ,
Walter Weder1, Isabelle Opitz1, Walter Klepetko4 , Nico Van
Zandwijk2, Glen Reid6
SWITZERLAND, 2Asbestos Diseases Research Institute, Sydney,
AUSTRALIA, 3Thoracic Oncology, Netherlands Cancer Institute,
Amsterdam, NETHERLANDS, 4Medical University of Vienna,
Vienna, AUSTRIA, 5Netherlands Cancer Institute, Amsterdam,
NETHERLANDS, 6Asbestos Diseases Research Institute, Sydney,
NSW, AUSTRALIA
Conclusion: Data from two independent validation series confirms the previously observed diagnostic potential of increased
miR-625-3p in blood from MPM patients. However, higher miR625-3p levels observed in NSCLC patients show that elevation
of the level of this microRNA in plasma/serum is not restricted
to MPM. The combination of cell-free microRNAs with SMRP
showed an improved diagnostic accuracy highlighting the need
for further investigation of multi-biomarker signatures.
Keywords: cell-free microRNAs, SMRP, diagnosis
1
Objectives: Definitive diagnosis of malignant pleural mesothelioma (MPM) is difficult and in most cases requires a tissue
biopsy of sufficient size. As a biopsy is not always feasible,
the identification of an accurate biomarker easily measured
in blood would represent an important step forward. The aim
of this study was to investigate the diagnostic accuracy of
combinations of soluble mesothelin-related protein (SMRP) and
cell-free microRNAs.
Methods: The study used two independent series of serum/
plasma samples: series 1 consisted of serum samples from
73 MPM patients, 69 healthy volunteers and 64 non-small cell
lung cancer (NSCLC) patients collected at the Netherlands
Cancer Institute between 1994 and 2013, and series 2 consisted of plasma samples from 36 MPM patients and 33 healthy
volunteers collected at the Medical University of Vienna and the
National Koranyi Institute of Pulmonology Budapest between
2011 and 2013. Cell-free miR-625-3p, and miR-126, previously
shown to have diagnostic potential were measured by RT-qPCR.
Additionally levels of soluble mesothelin-related protein (SMRP)
were assessed (ELISA) in the samples of series 1.
Results: Confirming previously published data, serum/plasma
miR-625-3p was found to be able to discriminate between MPM
and healthy controls with area under the ROC curve (AUCs) of
0.82 (95% CI: 0.75-0.89) in series 1 and 0.74 (95% CI: 0.630.86) in series 2. In addition, series 1 showed an AUC of 0.75
(95% CI: 0.67-0.84) for distinguishing NSCLC from healthy
controls and an AUC of 0.46 (95% CI: 0.47-0.66) for discriminating MPM and NSCLC. For miR-126 AUCs in series 1 were
at best 0.56 (95% CI: 0.47-0.66) for MPM vs healthy, while the
same comparison in series 2 resulted in an AUC of 0.76 (95%
CI: 0.64-.088). Assessment of SMRP in series 1 revealed AUCs
of 0.69 (95% CI: 0.59-0.78) differentiating MPM from healthy
individuals and 0.65 (95% CI: 0.54-0.75) separating MPM from
NSCLC, and an AUC of 0.64 (95% CI: 0.55-0.72) for the comparison MPM vs all non-MPM. Aiming to improve the diagnostic
accuracy, we evaluated the performance of combinations of
two or three of the tested biomarkers in series 1. This analysis
revealed the best performing combination for any comparison
to be miR-625-3p + SMRP which achieved AUCs of 0.87 (95%
CI: 0.81-0.92, MPM vs healthy), 0.63 (0.54-0.72, MPM vs NSCLC) and 0.76 (0.69-0.82, MPM vs all non-MPM), respectively.
Adding miR-126 to this combination did not improve diagnostic
accuracy.
PP01.20: TUMOR-INFILTRATING LYMPHOCYTES
AND BAP-1, VEGFR-2, IGF-1R EXPRESSION IN
Luca Ampollini1, Letizia Gnetti2, Matteo Goldoni3 , Nicoletta
Campanini2, Luigi Ventura1, Michela Solinas1, Cesare Braggio1,
Luigi Rolli1, Marcello Tiseo4 , Michele Rusca1, Paolo Carbognani1, Antonio Mutti3 , Enrico Maria Silini2
Department of Surgical Sciences, Thoracic Surgery, University
Hospital of Parma, Parma, ITALY, 2Pathological Anatomy and
Histology, University Hospital of Parma, Parma, ITALY, 3Department of Clinical and Experimental Medicine, University Hospital
of Parma, Parma, ITALY, 4Medical Oncology, University Hospital of
Parma, Parma, ITALY
1
disease strongly related to asbestos exposure. MPM is frequently associated with a prominent inflammatory reaction. In
this study, we investigated whether there was any relationship
between survival and the presence of tumor infiltrating lymphocytes (TILs). The expression of BAP-1 (BRCA1-Associated
Protein 1), VEGFR-2 (vascular endothelial growth factor receptor
2) and IGF-1R (Insulin-Like Growth Factor 1 Receptor) in tissue
samples was also assessed.
Methods: Forty-four cases of MPM were identified. All the biopsies used were obtained at the time of diagnosis. There were
24 males; mean age was 69 years. Twenty-six patients were
smokers and 26 had a certain history of asbestos exposure.
All histological slides were revised for the study; there were 28
epithelioid subtypes, 8 biphasic subtypes and 8 sarcomatoid
subtypes. The presence of TILs was scored as absent, weak,
moderate and strong according to a quantitative assessment
on hematoxylin and eosin slides. The expression of BAP-1,
VEGFR-2 and IGF-1R was analyzed by immunohistochemistry.
The impact of asbestos exposure, tobacco consumption and
histological subtypes on survival were also assessed. The survival analysis was analyzed by Kaplan Meier curve.
Results: TILs were present in 89% of cases (weak 18%, moderate 36%, strong 34%) and were found to be a favorable prognostic factor (p=0.05). Median survival in TILs and non-TILs
patients was 21 months and 4 months, respectively (Figure 1).
Epithelioid MPM was characterized by an increased expression
of VEGFR-2, both in tumor cells (p=0.04) and TILs (p=0.05) and
more frequent inactivation of BAP1 (75%, p=0.05) than biphasic
and sarcomatoid subtypes. IGF-1R was overexpressed in 57%
of the tumors (14 epitheliods and 11 sarcomatoids) and in 23%
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of TILs (7 epitheliods and 3 sarcomatoids). The expression of
VEGFR-2, BAP1 and IGF-1R was not significantly related with
survival. Tobacco (p=0.93) and asbestos (p=0.62) exposures
were also not significantly correlated with survival. Histology
had no effect on survival (p=0.23) although epithelioid MPM
fared better than sarcomatoid and biphasic tumors with a median survival of six months. work better in low-stage disease. Screening of asbestos exposed individuals by non-invasive methods is a desired path to
increase the rate of early detection and cure in the future.
Methods: Serum samples from the HUNT3 biobank, Levanger,
Norway were profiled with Magnetic Resonance Spectroscopy
(MRS metabolomics) and microRNA sequencing (Illumina).
The sample included mesothelioma cases collected 1-3 years
before diagnosis, along with age and sex-matched non-cancer
individuals; the never smokers to ever-smokers ratio was 1:2.
The produced data were analyzed with ad hoc bioinformatics
pipelines.
Results: There were six cases of pleural mesothelioma with
pre-diagnostic serum, three females and three males. Only one
was a never smoker while there was only one current smoker. Among the controls there were seven never smokers, two
current and three former smokers. All cases versus all controls
showed two metabolites significantly different by MRS, that was
overexpressed and down-regulated respectively (Fold change
1.33/0.62, False Discovery Rate adjusted p=0.01/0.001).
MicroRNA sequencing showed four significantly expressed microRNAs in serum, two overexpressed and two downregulated
(False Discovery Rate adjusted p‹0.05). However this signature
could not separate the groups completely. A 10-microRNA
signature did separate the two groups (Figure 1)
Conclusion: The presence of TILs favorably affects survival of
MPM. Epithelioid MPM is characterized by longer survival, higher BAP1 loss and increased VEGFR-2 expression. Histological
markers may improve the prognostic assessment of MPM and
provide mechanistic clues for new therapeutic strategies.
Keywords: VEGFR-2 (vascular endothelial growth factor receptor 2), IGF-1R (Insulin-Like Growth Factor 1 Receptor), tumor
infiltrating lymphocytes, BAP-1 (BRCA1-Associated Protein 1)
Conclusion: In this relatively small pilot study we identified
both metabolites and microRNAs that were significantly
differentially expressed in serum of mesothelioma patients 1-3
years prior to diagnosis. Importantly, we found that a 10-microRNA signature could differentiate the mesothelioma and
non-cancer group. These are preliminary results whose
validation in a larger cohort is still pending. There is hope that
significant biomarkers can soon be discovered for early
detection of mesothelioma within the metabolomics and
microRNA molecular spectre.
PP01.21: SERUM BIOMARKERS IN
MESOTHELIOMA 1-3 YEARS BEFORE DIAGNOSIS:
A PILOT STUDY
Robin Mjelle1, Maria Markaki2, Trygve Andreassen3 , Vincenzo
Lagani2, Ioannis Tsamardinos2, Tone F. Bathen3 , Pål Sætrom4 ,
Kristian Hveem5, Oluf D. Røe1
Department of Cancer Research And Molecular Medicine,
Norwegian University of Science and Technology, Trondheim,
NORWAY, 2Department of Computer Science, University of Crete,
Heraklion, GREECE, 3Department of Circulation And Medical
Imaging, Norwegian University of Science and Technology,
Trondheim, NORWAY, 4Department For Computer And Information Science, Norwegian University of Science and Technology,
Trondheim, NORWAY, 5Department of Public Health And General
Practice, Norwegian University of Science and Technology, Trondheim, NORWAY
1
Keywords: biobank, prospective study, early detection,
pre-diagnostic serum
Objectives: Early detection of mesothelioma could increase
survival and curation rate as low stage and low tumor burden
are positive prognostic factors. Moreover, emerging treatments
as immunotherapy and other biological treatments seem to
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PP01.22: ROLE OF EPHA2 IN MALIGNANT
MESOTHELIOMA
Yi-Hung Carol Tan1, Christopher Villaflor2, Brian M. Won1,
Rajani Kanteti1, Kyle Szeto2, Qudsia Arif3 , Aliya Husain3 , Wickii
T. Vigneswaran4 , Hedy Kindler1, Ravi Salgia5
Medicine, The University of Chicago, Chicago, IL, UNITED
STATES OF AMERICA, 2University of Illinos at Urbana-Champaign,
champaign, IL, UNITED STATES OF AMERICA, 3Department of Pathology, The University of Chicago, Chicago, IL, UNITED STATES
OF AMERICA, 4Loyola University Health System, Maywood, IL,
UNITED STATES OF AMERICA, 5City of Hope, Duarte, CA, UNITED
STATES OF AMERICA
1
Objectives: Overexpression of the ephrin-A1 ligand receptor
EPHA2 has been reported in many cancers and is associated
with tumor growth and metastatic potential. The role of EPHA2
in malignant mesothelioma (MM) remains unknown. We therefore investigated the expression and biology of EPHA2 in MM to
assess whether it could be an appropriate target for therapy.
Methods: To study the expression of EPHA2 in MM, immunohistochemistry and immunoblotting were performed on MM
patient samples and cell lines. Polymerase chain reaction (PCR)
and real-time PCR were used for gene mutation and amplification analysis, respectively. Doxazosin, a small molecule agonist
of the EphA2 receptor tyrosine kinase and cisplatin were used
in cell proliferation assay to study cell viability and wound healing assay was used for cell migration analysis.
Results: EPHA2 was over-expressed in 66.7% (4/6) of MM
cell lines. EPHA2 mutations were found in 28% (11/39) of MM
patients. EPHA2 gene amplification occurred in 10.3% (4/39)
of MM patient samples and in 33% (2/6) of MM cell lines. We
observed over-expression of EPHA2 in 64% (48/75) MM tumor
tissues with IHC staining score greater than 2+. Cells with
EPHA2 mutations had more cell proliferation and migration
than EPHA2 wild-type or normal controls. EPHA2 mutated cells
had greater resistance to cisplatin than EPHA2 wild-type cells
and one specific EPHA2 mutation showed resistance to both
doxazosin and cisplatin.
Conclusion: EPHA2 overexpression or mutation increased cell
proliferation and migration. EPHA2 mutation confers resistance
cisplatin. EPHA2 is a potential biomarker in this disease and
may be an appropriate target for MM therapy.
Keywords: EPHA2, target, biomarker, mesothelioma
PP01.23: ENOX2-BASED EARLY DETECTION OF
Jenette Creaney1, B Hosteler2, Dj Taggart2, Dm Morre2, Arthur
W. Musk3 , Bruce Robinson1, Dj Morre2
Asbestos Related Diseases, The University of Western Australia, Perth, WA, AUSTRALIA, 2MorNuCo Inc, Purdue University
Research Park, West Lafayette, IN, UNITED STATES OF AMERI1
CA, 3Respiratory Medicine, Sir Charles Gairdner Hospital, Perth,
WA, AUSTRALIA
Objectives: ENOX2 (Ecto-Nicotinamide Adenine Dinucleotide
Oxidase Disulfide-Thiol Exchanger 2) is a member of a family of
cell surface proteins that oxidize reduced pyridine nucleotides
[NAD(P)H] and are essential for cell enlargement and growth.
Recently specific isoforms of this protein have been found in
the serum of cancer patients before diagnosis. In this study we
explored if ENOX2 could be detected in mesothelioma patients
sera.
Methods: Archived serum samples from individuals exposed to
asbestos and who developed either benign disease or malignant
mesothelioma as determined from the Wittenoom Cancer Surveillance Program were assayed for the presence of mesothelioma-specific ENOX2 transcript variants using the ONCOblot
Tissue of Origin Cancer Detection Test, which employs 2-D gel
immuno- blot analysis of ENOX2 transcript variants in serum.
Results: Two mesothelioma-specific ENOX2 transcript variants
were detected in the serum of asbestos-exposed individuals 4
to 10 years prior to clinical diagnosis of malignant mesothelioma (average 6.2 years). Either one or both ENOX2 transcript
variants indicative of malignant mesothelioma were absent in
14 of 15 subjects diagnosed with benign pleural plaques either
with or without accompanying asbestosis.
Conclusion: In a population of asbestos-exposed subjects
who eventually developed malignant mesothelioma, ENOX2
transcript variants characteristic of malignant mesothelioma
were present in the serum 4 to 10 years in advance of clinical
symptoms.
Keywords: biomarker, ENOX2, early detection, serum
PP01.24: PROGNOSTIC MICRORNAS IN TISSUES
FROM MALIGNANT PLEURAL MESOTHELIOMA
PATIENTS RECEIVING MULTIMODALITY THERAPY
Michaela B. Kirschner1, Bart Vrugt2, Martina Friess1, Mayura
Meerang1, Peter Wild2, Nico Van Zandwijk3 , Glen Reid3 , Walter
Weder1, Isabelle Opitz1
SWITZERLAND, 2Institute of Surgical Pathology, University Hospital Zurich, Zurich, SWITZERLAND, 3Asbestos Diseases Research
Institute, Sydney, NSW, AUSTRALIA
1
Objectives: In a recent study using two cohorts (N=48 and
N=43) of surgical specimens from malignant pleural mesothelioma (MPM) patients, we identified several microRNAs with
prognostic potential. A combination of six of these microRNAs,
which we termed the miR-Score, provided the best prognostic
accuracy for identifying patients with a survival of >20 months
following surgery. The purpose of the current study is to further
evaluate these microRNAs in surgical and diagnostic tumor
specimens of an independent cohort of MPM patients receiving
multimodality treatment.
Methods: We identified a cohort of 140 MPM patients who
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ABSTRACT BOOK
received chemotherapy followed by extrapleural pneumonectomy (EPP) between 1999 and 2014 at the University Hospital
Zurich. At present microRNA analysis has been carried out in a
small subset of 25 EPP (all chemo-pretreated) and 13 matching
diagnostic (chemo-naïve) specimens. RNA was isolated from
microdissected tumor tissue, and then subjected to microRNA specific RT-qPCR for the six microRNAs of the miR-Score.
Kaplan-Meier log rank analysis was performed to determine
association of the microRNAs with survival. Related-samples
Wilcoxon Signed Rank Test was employed to determine differences in microRNA expression between diagnostic and surgical
specimens.
Results: For five of the six signature microRNAs Kaplan-Meier
analysis in both EPP and diagnostic specimens showed similar
associations between expression levels and survival as previously published. However, miR-21, the microRNA previously
most significantly associated with prolonged survival, did not
reach statistical significance in the current subset (p=0.764).
As a result of the analysis of the individual microRNAs, also
the combined miR-Score was not significantly associated with
survival. This is most likely attributable to the small number of
samples currently available. To evaluate whether chemotherapy
might affect microRNA levels, we compared expression before
and after chemotherapy in 13 matched sample pairs. While
this analysis did not show significant differences in the median
expression between pre- and post-chemotherapy tissue for
any of the microRNAs, on the sample-by-sample investigation
showed reduction in expression of miR-21 after chemotherapy.
For all other microRNAs, while individual samples did show
strong differences between pre- and post-chemo expression, no
conclusive trend regarding the effect of chemotherapy could be
observed.
Conclusion: Preliminary data from an independent series of
MPM specimens (diagnostic and post chemotherapy) show that
despite the small number of investigated samples, the majority
of microRNAs with prognostic potential could be validated. Our
data also suggest that miR-21 levels may be affected by chemotherapy. Ongoing investigation in a larger number of samples is
needed to validate the miR-Score and to provide more insight
into the effect of chemotherapy on microRNA expression, and
how this might be associated with response to therapy.
Keywords: multimodality treatment, prognosis, microRNA
PP01.25: CHEMOTHERAPY AND CHEMOTHERAPY
PLUS TALC PLEURODESIS IN TREATMENT OF
MALIGNANT PLEURAL MESOTHELIOMA: A
RETROSPECTIVE ANALYSIS
Guntulu Ak1, Muzaffer Metintas1, Huseyin Yildirim1, Selma
Metintas2, Ulku Yilmaz3 , Cansel Atinkaya Ozturk4 , Isa Dongel5,
Gokturk Findik6 , Senay Yilmaz1, Akın Ozturk7, Elcin Ersoz4 ,
Derya Kızılgöz3 , Pınar Akın Kabalak3 , Tuba Inal Cengiz3
Lung And Pleural Cancers Research And Clinical Center And
Medical Faculty Department of Chest Diseases, Eskisehir Osmangazi University, Eskisehir, TURKEY, 2Lung And Pleural Cancers
1
Research And Clinical Center And Medical Faculty Department
of Public Health, Eskisehir Osmangazi University, Eskisehir,
TURKEY, 3Pulmonary Oncology Unite, Ataturk Chest Diseases and
Chest Surgery Training and Educational Hospital, Ankara, TURKEY, 4Department of Chest Surgery, Sureyyapasa Chest Diseases
and Chest Surgery Training and Educational Hospital, İstanbul,
TURKEY, 5Medical Faculty Department of Chest Surgery, Suleyman Demirel University, Isparta, TURKEY, 6Department of Chest
Surgery, Ataturk Chest Diseases and Chest Surgery Training and
Educational Hospital, Ankara, TURKEY, 7Department of Pulmonary Oncolgy Unite, Sureyyapasa Chest Diseases and Chest
Surgery Training and Educational Hospital, İstanbul, TURKEY
Objectives: Surgery combined with radiotherapy and/or chemotherapy have been reported to improve median survival in
patients with malignant pleural mesothelioma (MPM). However,
since the majority of patients are ineligible for or refuse surgery,
chemotherapy represents the only therapeutic modality in such
cases. There has been a recent interest in the use of therapeutic options that can be used as adjunct to chemotherapy
in MPM patients. Among those, talc pleurodesis administered
in to pleural fluid for palliative purposes has been proposed to
improve MS in MPM. The present study aimed at retrospectively
assessing the effect of talc pleurodesis on MS in a group of
MPM patients undergoing chemotherapy.
Methods: A total of 248 patients ineligible for or refusing
surgery who were treated with chemotherapy and who were
followed-up for a minimum duration of 12 months between
January 1991 and June 2015 were included. Age, gender, tumor
cell type, disease stage, treatments administered, time of diagnosis and initiation of treatment, response to chemotherapy,
KPS, outcome of pleurodesis, treatment related side effects,
and death certificates were retrieved from the medical database. Patients with missing data or with a follow up duration of
less than 12 months were excluded. Patients were divided into
the two following groups: “chemotherapy alone” or “chemotherapy plus talc pleurodesis”. The two groups were compared with
respect to factors that may have an influence on the prognosis
of MS such as age, gender, stage, KPS, tumor histology, and
treatment modality.
Results: Of the 248 total participants, 134 were male and 114
were female with a mean age of 61.3 years (60.7 in males, and
62.2 in females). Chemotherapy alone was administered in 152
patients, while talc pleurodesis was given additionally in 96.
Histological diagnoses were epithelioid, myxoid, and sarcomatoid in 193, 33, and 10 patients, respectively, while histological type could not be ascertained in 12 patients. There were
19, 42, 96, and 91 patients with stage 1, 2, 3, and 4 disease,
respectively. A total of 155 patients had a KPS of ≤ 80, while
93 had a KPS > 80. The treatment groups were comparable
in terms of gender (p=0.578), distribution of histological cell
types (p=0.604), stage (p=0.669), and KPS (p=0.224). The MS
in the overall patient group, i.e. 248 patients, was 12 months,
while the corresponding figures were 13 and 9 months for
epithelioid and non-epitheloid tumors, respectively (p<0.001 ).
Again MS in stage 1, 2, 3, and 4 patients were 16, 15, 13, and 9
months respectively (p<0.001). Duration of MS was 10 months
and 17 months in those with a KPS of ≤ 80 and > 80, respectively (p<0.001). Chemotherapy alone and chemotherapy plus
talk-pleurodesis were associated with respective MS of 11 and
13 months (p=0.8884). Logistic regression analysis showed that
male gender (p=0.034), advanced stage (p=0.002), non-epitheloid histology (p=0.001), and low KPS (p<0.001) were assoiMig2016.ORG
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ABSTRACT BOOK
ciated with a poorer MS, with no effect of the addition of talk
pleurodesis to chemotherapy on MS (p=0.391). When considering patients with epithelioid tumors only, stage (p=0.007) and
KPS (p<0.001) emerged as significant predictors of MS, while
addition of talc pleurodesis did not appear to have an effect on
MS (p=0.660). In both treatment groups, the side effects were
generally tolerable and no treatment-associated deaths were
observed.
Conclusion: In this retrospective analysis of a limited number
of patients, addition of talc pleurodesis to chemotherapy did
not result in a significant improvement in MS in patients MPM,
although the survival difference between the two groups was 2
months. Thus, a prospective randomized study conducted by
the Turkish Mesothelioma Working Group is currently underway
to further assess the role of talc pleurodesis in these patients.
*This study is supported by General Directorate of Health Researches, Republic of Turkey, Ministry of Health.
Keywords: prognosis, mesothelioma, Talc pleurodesis, chemotherapy
PP01.26: INITIAL RESULTS OF THE MULTICENTRIC
TURKISH MALIGNANT PLEURAL MESOTHELIOMA
DATABASE*
Hasan Batirel
Thoracic Surgery, Marmara University Faculty of Medicine, Istanbul, TURKEY
Objectives: Malignant Pleural Mesothelioma (MPM) is a
disease with poor prognosis. Environmental and occupational
asbestos exposure are the main factors for MPM. We started
an online MPM database in 2014. In this study we analyzed the
initial results of this database.
Methods: During 2014-2015, cases with a histologic diagnosis of MPM were recorded in an online MPM database by the
Turkish Mesothelioma Study Group. 301 cases were recorded.
Dual submissions, patients without a date of diagnosis, followup
data and date of diagnosis earlier than 1.1.2013 were excluded.
Eventually 204 cases were evaluable. Demographic, histologic,
clinical and pathologic stage, treatment and survival data were
analyzed. Kaplan-Meier survival and uni-, multivariate analyses
were performed.
Results: Average age was 62 ± 11 (76 females). Histologies
were epithelioid (n=136), biphasic (n=34), sarcomatoid-desmoplastic (n=24) and subtype not identified (n= 9). Clinical T
and N stage was available in 124 and 52 patients respectively.
Pathologic T and N stage was available in 54 and 36 patients
respectively. 80 patients underwent treatment that included
surgery. Mean follow-up was 12.4 ± 8.1 months. Overall median
and 2 year survivals were 14.6 months and 35% respectively.
Clinical M1 disease patients (n=18) had a median survival of
7.1 months. There was no difference in median survival due
to gender or side (Male/Female 14.6/14.5 months, p=0.92;
Right/Left 13.4/18.5 months, p=0.51). Survival comparisons
are shown in Table 1. Histology, Clinical T and N stage and
treatment were significant (p<0.001, p=0.005, p=0.002 and
p<0.001 respectively). Multivariate analyses showed histology
(p=0.003, OR 8.7) and type of treatment (p=0.002, OR 9.5) as
significant factors. Criteria (n)
Median
Survival
(months)
2-yr
Survival
(%)
p-Value
Epithelioid (136)/
Biphasic (34)/
Sarcomatoid (24)
18.5/13.3/
7.1
42/22/0
<0.001
Asbestos History
Present (n=106)
/Absent (n=98)
12.9/18.5
Clinical
T1(n=8)/2(n=44)
/3(n=36) /4(n=36)/
X(n=80)
12.9/19.8/
18.8/8.8/
16.8
29/37/
42/11/43
0.005
Clinical N0(n=32)
/2(n=20)/X(n=152)
24.9/8.1/
14.3
56/0/33
0.002
Pathologic
T1(n=5)/2(n=15)/
3(n=27)/4(n=7)
16.9/16.1/
21.2/NR
Treatment Including
Surgery (n=80)/
Without Surgery
(n=91)/
Supportive (n=27)
19.8/12.4/
4.2
46/28/19
<0.001
Surgery with MCR
(n=51)/
Without MCR(n=29
21.2/15.8
50/47
0.58
32/40
0.093
0.39
Conclusion: Turkish MPM cohort which is mainly based on
patients with environmental asbestos exposure with similar
demographic characteristics and survival rates as in literature.
Treatments that include surgery were associated with a prolonged survival which is likely due to a selection bias. Histologic
subtype identification is very important for prognostic stratification. Turkish Mesothelioma Working Group Contributors:
Muzaffer Metintas, Hasan Fevzi Batirel, Sedat Altin, Cansel
Atinkaya Ozturk, Berna Oksuzoglu, Ulku Yilmaz, Figen Deveci, Adil Zamani, Adnan Sayar, Nazan Sen, Serdar Berk, Ufuk
Yilmaz, Dilek Ernam, Pınar Akin Kabalak, Volkan Kara, Derya
Ozaydin, Mehmet Ali Bedirhan, Tuba İnal Cengiz, Ibrahim
Dincer, Isa Dongel, Talat Kilic, Zehra Seyfikli, Mehmet Bayram,
Erdogan Cetinkaya, Ali Kadri Cirak, Gamze Kirkil, Celalettin
Kocaturk, Muzaffer Metin, Berna Akinci Ozyurek, Umran Toru. *
This study was was supported by General Directorate of Health
Researches, Ministry of Health, Republic of Turkey.
Keywords: chemotherapy, Survival, Surgery, mesothelioma
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ABSTRACT BOOK
PP01.27: FOURTH LINE CHEMOTHERAPY
WITH DOCETAXEL FOR MALIGNANT PLEURAL
MESOTHELIOMA (MPM)
PP01.28: PHASE I TRIAL OF LENTIVIRAL CARTMESO CELLS IN PROGRESSIVE MALIGNANT
PLEURAL MESOTHELIOMA (MPM)
Jens B. Sørensen, Vinicius De Lima
Andrew R. Haas, Gabriela Plesa, Drew A. Torigian, Edmund
Moon, Mark O’Hara, Gregoary Beatty, Janos L. Tanyi, Annemarie Nelson, Naseem Kerr, Maureen Mcgarvey, Simon Lacey, Jos
Melenhorst, Steven Albelda, Carl June
Dept. Oncology, Finsen Centre/National University Hospital,
Copenhagen, DENMARK
Objectives: Docetaxel has revealed activity in malignant pleural mesothelioma (MPM) with response rates from 5%-23%
when used as 1st line treatment (Vorobiof DA et al. 2002, Belani
CP 2004, Sørensen JB 2008). There are no well-defined 3rd- or
even 4th-line treatments for advanced MPM, but we use routinely carboplatin/gemcitabine/liposomized doxorubicin (CCG
regimen) as 3rd line because of the activity previously reported
(de Lima, 2015. Hence, Docetaxel was explored as a possible
4th line in MPM.
Methods: Patients had histologically verified MPM, progression after 1st line platinum-pemetrexed, 2nd line vinorelbine,
and 3rd line carboplatin/gemcitabine/liposomized doxorubicin
(CCG regimen), and PS 0-2. The explored 4th line regimen was
docetaxel 75 mg/m2 day 1 q3wks, maximum 6 cycles. CT-scans
were done after every second cycle.
Results: Out of 564 MPM patients who received 1st line
platinum/pemetrexed 2010-2015 13 patients (2.3%) received
later 4th line docetaxel. These 13 patients had median age 65
years, 92% were males, 15% PS 2, 46% epitheloid subtype
and 54% biphasic, and all had stage IV disease. Median time
from diagnosis to start of 4th line docetaxel was 21.5 months.
31% of patiens had previous palliative radiotherapy and 39
had palliative pleurectomy. Median treatment duration was
2 cycles (range 1-6 cycles). Treatment was postponed due to
hematologic toxicities in 8% and dose reduction was necessary
in 5 paatients (39%)(3 pts due to hematologic toxicity, one due
to neutoxicity and one of other causes). A total of 4 CTC grade
3 events occurred (allergic reactions 2 cases, anemia 2 cases)
and one grade 4 event (neutropenia). No toxic deaths occurred.
There were no objective responses and disease control rate
(DCR) was 23%. Medians of progression free survival (PFS) and
OS were 1.5 and 4.6 months from start of 4th line treatment,
respectively.
Conclusion: 4th line treatment with docetaxel could be safely
administered to heavily pretreated advanced MPM starting
nearly 2 years after initial diagnosis. However, docetaxel did not
have noteworthy activity in this late stage of the disease. More
efficacious salvage regimens are sorely needed.
Keywords: Docetaxel, 4th line treatment, chemotherapy
AMERICA
Objectives: Epithelial MPM expresses the tumor antigen
mesothelin. Due to mesothelin’s limited expression profile in
normal tissues, it is a potential ideal target for mesothelin-directed cellular-based immunotherapies. We performed a phase
I clinical trial of lentiviral CART-meso cells in patients with
progressive MPM.
Methods: Five patients with epithelial MPM underwent a 10
liter apheresis for peripheral blood mononuclear cell isolation.
Ex vivo transfection with a lentiviral chimeric anti-mesothelin
immunoreceptor SS1 fused to the 4-1BB and CD3ζ signaling
domains was performed.
In a standard 3+3 phase I dose escalation format, patients
received 1e7 or 1e8 CART-meso cells intravenously without or
with 1.5 gm/m2 cyclophosphamide as a conditioning regimen
two to four days prior to CART-meso cell infusion. Safety was
assessed via standard CTCAE patient assessment and biochemical evaluation. A variety of immunologic and biologic correlates
were assessed including CART-meso cell persistence and
trafficking, human anti-mouse/CAR antibody (HAMA/HACA)
development, and modified RECIST criteria response.
Results: CART-meso cells without or with cyclophosphamide
were administered without any evidence of on-target off-tumor
toxicity to pleura, peritoneum or pericardium. One serious
adverse event occurred in the cyclophosphamide group due to
neutropenic fever requiring hospitalization for intravenous antibiotics. By Q-PCR, CART-meso RNA peaked between day 10-14
then trended toward undetectable by day 28. As expected, a
nearly 10-fold higher CART-meso RNA level was detected at day
10-14 at 1e8 cells compared to 1e7 cells. Moreover, at both 1e7
and 1e8 cells, cyclophosphamide increased RNA level 10-fold
over the dose without cyclophosphamide. One of the 5 patients
infused with CART-meso cells had malignant ascites which was
tapped post CART-meso cell infusion. CART-meso construct
was detected in the ascites at days 9 and 17, but not day 28. Interestingly, this patient also had the highest level of mesothelin
surface and cytoplasmic expression. Low level HAMA expression was present in one patient and interestingly the patient
with malignant ascites had detectable HACA in their peritoneal
fluid. By modified RECIST criteria, 4 of 5 patients had stable
disease at one month, but all patients had progressive disease
by 3 months. The patient with malignant ascites remains alive
10 months post CART-meso infusion on gemcitabine therapy.
Conclusion: We were able to demonstrate the safety of a
lentiviral murine CART-meso cellular therapy without and with
cyclophosphamide in MPM patients. CART-meso cells demonstrated expansion and persistence that was enhanced with
cyclophosphamide conditioning and trafficking was detected in
the one patient. Despite no dramatic responses, these results
demonstrate safety regarding no on-target off-tumor toxicity
and a fully human CART-meso construct is in development
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ABSTRACT BOOK
and awaits a phase I trial to evaluate safety, biocorrelates and
efficacy. Ultimately, combinatorial therapies to improve trafficking and to modify the tumor microenvironment may further
enhance CART cell therapies in MPM and other solid tumors.
Keywords: Cyclophosphamide, T cells, chimeric antigen receptor, phase I trial
PP01.30: MESOTHELIOMA MORTALITY IN
AUSTRALIAN AND ITALIAN-BORN WORKERS
EXPOSED TO BLUE ASBESTOS AT WITTENOOM,
WESTERN AUSTRALIA
Nicholas De Klerk1, Alison Reid2, Enzo Merler3 , Susan Peters4 ,
Peter Franklin4 , Vittoria Bressan3 , Arthur W. Musk4
Telethon Kids Institute, University of Western Australia, Subiaco,
WA, AUSTRALIA, 2Curtin University, Perth, WA, AUSTRALIA, 3Mesothelioma Register of the Veneto Region, Padua Local Health
Unit, Padova, ITALY, 4School of Population Health, University of
Western Australia, Perth, WA, AUSTRALIA
1
PP01.29: DOES MALIGNANT PLEURAL
MESOTHELIOMA WITH METASTASES AT
FIRST PRESENTATION REALLY BENEFIT FROM
CHEMOTHERAPY?
Hala Aziz Shokralla1, Mohamed Rahouma2
Medical Oncology, national cancer institute, cairo, EGYPT, 2Surgical Oncology, national cancer institute, cairo, EGYPT
1
Objectives: Our study aims to evaluate different clinico-pthological characteristics of metastatic cases of MPM at presentation and the possible benefits from active therapy from a single
Institution practice data.
Methods: We retrospectively analyzed patients with locally
advanced or metastatic MPM, treated at the Department of
Medical Oncology –national cancer institute in Egypt. Data
on age, gender, smoking history, asbestos exposure, performance status, tumor stage, histology, type of treatment
(Raltitrexed-Bortezomib-Gemcitabine-Vinorelbine with platinum
containing agents), response to treatment and routine laboratory tests including complete blood count panel, date of death or
censored status were collected. Mean Progression free survival
and overall survival were estimated.
Results: 114 patients had MPM. Fifteen cases present as
metastatic disease (13%). Nine patients (60%) were men. The
median age of patients was 52 years (range; 19-73) and mean
pre-treatment weight was 72.6 kg. All cases were in performance status –I at presentation apart of five cases (two with
PS-II and three with PS-III). Thirteen (87%) patients reported
asbestos exposure. Dyspnea and Chest pain were the most
prevalent symptoms (94%). Only one case had no pleural
effusions at presentation, thickening was obvious in all cases,
five cases had no mediastinal nodes (~34%) Eight cases had
osseous deposits, five had hepatic focal lesions, two had
contralateral lung metastasis, only one case had axillary nodes
and another one had malignant ascites. Twelve cases received
platinum containing combination, eight cases experienced
progressive disease (~67%). The mean overall survival (OS)
and Progression free survival (PFS) were 13.6 months and 7.8
months, respectively.
Conclusion: Our results suggest that mesothelioma may
present rarely with wide spread disease; most of them didn’t
respond to platinum containing therapy. Further studies may
confirm possible benefit of chemotherapy compared to supportive therapy alone.
Objectives: To compare mortality from malignant mesothelioma and other diseases among Italian-born and Australian-born
workers employed at Wittenoom, Western Australia, using rates
from the general Western Australian and Italian populations.
Methods: The minesite operated from 1943 and 1966 and
work histories were based on employment records from Australian Blue Asbestos (ABA) who operated the Wittenoom mine.
There were over 50 different nationalities recorded with Italians
forming the largest migrant group. British and Australians were
similarly recorded and not distinguishable from each other.
Italians were identified both through company records and subsequent verification of all records and attribution on the basis
of full name. Follow-up from 1943 to 2009 was done in both
Australia and Italy for death and cancer incidence. Asbestos
exposure was based on duration of employment and various
industrial hygiene surveys which allocated specific levels to
different jobs. No information on overtime or extra work periods
was available. SMRs were based on both Australian and Italian
rates and Cox regression models were used to examine the
separate and combined effects of exposure and ‘nationality’ on
mesothelioma mortality.
Results: The mesothelioma mortality rate, per 100,000, was
higher among the Italian-born workers (181.4, 95%CI 145.7225.8) than the Australian/UK-born workers (119.6, 95%CI
103.2-138.5). Within both groups of workers the risk of mesothelioma increased with an increasing level of cumulative exposure. Comparing the risk between Italian and Australian/UKborn workers by category of exposure showed a greater than
twofold increased risk of mesothelioma among Italian workers
in the lowest exposure category (<10 f/ml years), compared
with Australian/UK-born workers. The risk of mesothelioma was
not statistically different between Italian or Australian/UK-born
workers in either the medium (10-50 f/ml years) or high (>50 f/
ml years) exposure categories.
Conclusion: 30% of Italian immigrants who left Wittenoom
have returned to Italy, most after at least two years of migration. Their time at Wittenoom has deeply affected their health
with high risks for asbestos-related disease. Mortality from
asbestos disease was significantly more marked in the Italians
than in the British/Australians, and is perhaps indicative of
discrimination against migrants.
Keywords: crocidolite, migrant health, Malignant mesothelioma
Keywords: metastatic-MPM-presentation-therapy
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PP01.31: PRELIMINARY REPORT OF AN
OBSERVATIONAL CLINICAL REGISTRY ON
MALIGNANT PLEURAL MESOTHELIOMA (MPM) IN
ITALY (REGCLIN)
Federica Grosso1, Annalisa Roveta2, Paolo Pedrazzoli3 , Francesco Valentino3 , Daniela Degiovanni4 , Alessandra Bearz5,
Federico Rea6 , Francesco Cognetti7, Armando Santoro8 , Tiziana
Cena9, Alberto Muzio10, Gianmauro Numico1, Carmine Pinto11,
Massimo D’Angelo12, Giorgio V. Scagliotti13 , Corrado Magnani9
Oncology Unit, SS. Antonio e Biagio e C. Arrigo, Hospital, Alessandria, ITALY, 2Ssa Sviluppo E Promozione Scientifica, SS. Antonio e Biagio e C. Arrigo, Hospital, Alessandria, ITALY, 3Oncology
Unit, Fondazione IRCCS Policlinico S. Matteo, Pavia, ITALY, 4Ss
Hospice Monsignor Zaccheo/ Uocp, Santo Spirito, Hospital,
Casale Monferrato, ITALY, 5Oncology Unit, Centro Riferimento Oncologico IRCCS, Aviano, ITALY, 6Divisione E Cattedra Di Chirurgia
Toracica E Centro Trapianto Di Polmone, Azienda Universitaria
Ospedaliera, Padova, ITALY, 7Oncology Unit, Istituto Nazionale
Tumori IRCCS Regina Elena, Roma, ITALY, 8Oncology Unit, Istituto
Clinico IRCCS Humanitas, Rozzano, ITALY, 9Medicina Traslazionale, Università degli Studi del Piemonte Orientale, Novara, ITALY, 10 Oncology Unit, Santo Spirito, Hospital, Casale Monferrato,
ITALY, 11Oncology Unit, Azienda Ospedaliera Arcispedale - IRCCS
Santa Maria Nuova, Reggio Emilia, ITALY, 12Centro Sanitario
Amianto, Azienda Sanitaria Locale AL, Casale Monferrato, ITALY, 13Dipartimento Di Oncologia Medica, Università di Torino-San
Luigi Hospital, Torino, ITALY
Acknowledging the current incompleteness of the data it can
be currently extrapolated that, a high proportion of patients
received active treatment and a significant percentage of them
was included in experimental studies. The overall survival
compares favorably with historical data. While continuing to
register new cases, we are now planning to expand this project
also through the integration with the epidemiological registry
ReNaM (Registro Nazionale Mesotelioma).
Keywords: Malignant pleural mesothelioma, Registry, Database, Observational study
1
Objectives: The present study, supported by an Italian CCM
(Centro per il Controllo delle Malattie) project , provides information about recruitment, clinical characteristics, treatment
modalities, and outcomes of a large series of MPM patients
included in the Italian observational protocol REGCLIN. Data
were collected in a web-based registry of patients treated in 10
participating referral Institutions. Here we report on the preliminary analysis of the data collected so far.
Methods: The most relevant clinical and treatment related
variables for each patient were registered in the web database
from each participating center. The collected data was then
analyzed using SAS (9.2 v.).
Results: Since January 2010 to November 2015, 359 MPM patients, 249 males (69%) and 110 females (31%), with a median
age 70 (IQR 64-77; range 29-90) were included. Hystology was
epithelioid in 261 (73%), biphasic in 49 (14%), sarcomatous in
36 (10%) cases and missing in 13 (3%). Diagnosis was obtained
through pleuroscopy/thoracoscopy in 304 (85%) and through
CT guided biopsy in 31 (9%), and through other modalities in 24
(6%) cases. Data about treatment modalities were available for
277 (77%) patients: 262 (94%) received chemotherapy, representing the only treatment modality for 163 (59%). Surgery was
performed in 71 (26%) patients, always associated with chemotherapy; of these, 37 (13%) received multimodal approaches
including also radiotherapy. Sixty-nine patients (25%) were
treated in the context of clinical trials. The follow-up analysis is
ongoing: it is currently available for 258 (72%) patients and, to
date, overall survival (OS) is 15,7 months (IC90 13,8 – 18,3).
Conclusion: This study demonstrates the feasibility of a webbased multi-institutional clinical and pathological registry that
allows data sharing about diagnosis and treatment of MPM.
PP01.32: SLOWLY PROGRESSIVE MALIGNANT
PLEURAL MESOTHELIOMA IN AGED PATIENTS
Mizue Hasegawa1, Asako Okabayashi1, Akitoshi Sato1, Naoko
Yokohori1, Hideki Katsura1, Toshiko Kamata2, Eitetsu Koh2,
Yasuo Sekine2, Di Wu3 , Kenzo Hiroshima3
Respiratory Medicine, Tokyo Women’s Medical University,
Yachiyo Medical Center, Yachiyo, JAPAN, 2Thoracic Surgery, Tokyo
Women`s Medical University, Yachiyo Medical Center, Yachiyo,
JAPAN, 3Pathology, Tokyo Women`s Medical University, Yachiyo
Medical Center, Yachiyo, JAPAN
1
Objectives: Malignant pleural mesothelioma (MPM) predominantly affects men aged 50-70 years. Without treatment, it is
associated with a poor median survival, ranging from 4 to 12
months. Management for the aged patients is not fully elucidated since invasive examinations or treatments may be intolerable. We retrospectively evaluated the clinical and pathological
features of MPM in aged patients diagnosed at our institution.
Methods: Two patients were diagnosed as MPM over the age
of eighty from March 2010 to December 2015 at our institution.
Medical records were analyzed retrospectively.
Results: Case 1; A 90 year-old man admitted to our hospital for
fatigue and appetite loss. Chest X-ray and computed tomography (CT) revealed massive right pleural effusions and bilateral
pleural plaques without remarkable thickening of pleura or
mass lesion. Examination of pleural effusion showed elevation
of hyaluronic acid with 170,000ng/ml, but cytological examination could not address diagnosis of MPM. After drainage of
pleural effusion, palliative care was continued without assertive examination or treatment. After 8 months, patient died of
pneumonia without increase of pleural effusion or progression
of pleural lesions. Autopsy revealed slightly thickened pleura
without obvious mass lesion. Microscopic findings of pleura
revealed proliferation of round to oval cells with acinar formation. Immunohistochemical stainings were positive for CAM5.2,
carletinin, D2-40, and negative for CEA, TTF-1.Final diagnosis
was epithelioid MPM. Case 2; An 87 year-old man admitted
to our hospital with hydro-pneumothorax. Chest CT revealed
right hydro-pneumothorax with minimal pleural thickening or
mass. Cytological examination of pleural effusion could not
address diagnosis of MPM. Video-assisted thoracic surgery
was performed for the treatment of refractory pneumothorax.
Pathological findings of pleura revealed proliferation of round to
oval cells with tubulopapillary pattern. Immunohistochemical
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stainings were positive for carletinin, D2-40, and negative for
CEA, TTF-1. Final diagnosis was epithelioid MPM. Palliative care
was continued until 5 months after when he died of pneumonia
without marked progression of MPM lesions.
and 31/12/2005, therefore MPM account for a total of 0.19%
of all cases. It seems that second and third lines are useless.
New studies and/or new drugs are mandatory for this disease
category.
Conclusion: We presented 2 cases of MPM in aged. Both patients were epithelial type with slow progression of MPM lesions
and died of disease other than MPM. Invasive examination or
treatment may not be helpful for aged patients with suspect or
under diagnosis of MPM.
Keywords: mesothelioma, asbestos
Keywords: mesothelioma, aged, slow progression
PP01.34: BRAIN METASTASES IN MALIGNANT
PP01.33: A COHORT OF MALIGNANT PLEURAL
MESOTHELIOMA TREATED AND FOLLOWED IN
THE LAST TEN YEARS AT INSTITUTO ONCOLÓGICO
HENRY MOORE
Mariana Abal, Ernesto Gil Deza, Claudia Acuña, Carlos Garcia
Gerardi, Gabriela Malcervelli, Dario Niewiadomsky, Flavio
Tognelli, Eduardo Morgenfeld, Felipe Gustavo Gercovich
Instituto Oncologico Henry Moore, Buenos Aires, ARGENTINA
Objectives: The main objective of this paper is to analyze the
clinical features and outcome of pt diagnosed with MPM who
were treated at our institution during the last 10 years.
Methods: We reviewed the clinical records of 50 pt diagnosed
with MPM between 2005 and 2015. All the pt were treated and
followed at our institution from the diagnosis. We reviewed the
occupational exposure to asbestos, personal and family history,
comorbidities, tumor stages, PS at presentation, pharmacological and/or invasive interventions. The tumor response was evaluated according to RECIST 2.0. Overall survival was registered
from diagnosis until death or to the last pt’s visit.
Results: Fifty pt were studied, the average age was 65 years
old (range 44-85). Sex: male 28 pt (56%) / female 22 pt (44%).
Occupational asbestos exposure was confirmed in 3 pt and suspected in 9 pt. No pt had a history of relatives diagnosed with
MPM. Smoking was present in 22 pt. Stages: I: 3 pt (6%) , II: 7
pt (14%) , III: 18 pt (36%) and IV: 22 pt (54%). ECOG: 0-1: 27 pt
(54%) , 2 : 20 pt (40%) , 3 :3 pt (6%) .In 45 pt (90%) there were
no other concomitant neoplasias. The most frequent histologic
subtype was Epithelioid in 36 pt (72%). Five pt underwent a
surgical procedure (1 pt decortication ; 1 pt pleurectomy and
pericardiectomy ; 1 pt tumor resection from chest wall and 2 pt
pleurodesis ). Radiotherapy was indicated in 3 pt with palliative
intent. Chemotherapy was the first therapeutic treatement
in 40 pt (80%) . All pt received a first line of Platinum and
Pemetrexed, 12 pt received a second line with gemcitabine or
vinorelbine, only one pt received a third line. Eight out of 40 pt
(20%) treated with the first line had a partial response and they
had a median survival of 20 months (4-64) . No pt responded
to the second line of treatment. Median overall survival for the
total cohort was 11.3 months (range 1-64 m).
Nobukazu Fujimoto1, Tomoko Yamagishi2, Yosuke Miyamoto2,
Michiko Asano2, Yasuko Fuchimoto2, Sae Wada2, Shinji Ozaki2,
Hideyuki Nishi2, Takumi Kishimoto2
Medical Oncology, Okayama Rosai Hospital, Okayama, JAPAN, 2Okayama Rosai Hospital, Okayama, JAPAN
1
Objectives: The brain is a rare site of metastasis in malignant
pleural mesothelioma (MPM), and its clinical features and prognosis remain unclear. The aim of this study was to investigate
the incidence, prognosis, and risk factors for brain metastases
(BM) in MPM patients.
Methods: The study included 150 consecutive patients with
histologically proven MPM who were seen between July 1993
and October 2014 at Okayama Rosai Hospital, Japan. Baseline
demographic and clinicopathological variables were collected
retrospectively from patients’ medical records. These included
age at initial diagnosis, gender, histological subtype, clinical
stage, and baseline Eastern Cooperative Oncology Group
(ECOG) performance status (PS).Diagnosis of BM in MPM was
based on magnetic resonance imaging (MRI) or contrast-enhanced computed tomography (CT) scans. Routine brain imaging was performed at the diagnosis but not during the follow-up
period, unless BM was suspected. Leptomeningeal metastases
were not included in the actuarial incidence of BM in this study.
Results: The median follow-up time was 11 months (range
0–154.0 months). A total of eight patients (5.3%) developed
BM during the course of their illness. Multivariate analysis
identified age < 65 years (odds ratio [OR] = 5.83, p = 0.038)
and International Mesothelioma Interest Group stage IV (OR
= 1.69, p = 0.040) as independent factors related to increased
risk of developing BM. The 1-and 2-year cumulative rates of BM
were 4.0% (95% confidence intervals [CI] 1.4-8.5%) and 5.3%
(95%CI 2.3–10.2%), respectively. Our study showed that the
overall survival (OS) of patients with BM was worse than that of
patients without BM (median OS 6.5 versus 11.0 months, p =
0.037).
Conclusion: The prognosis for BM in MPM patients is poor.
Clinicians should perform careful screening for BM, especially
in patients with risk factors.
Keywords: Brain metastases, Malignant pleural mesothelioma,
asbestos
Conclusion: The Instituto Oncológico Henry Moore in Buenos
Aires, Argentina registered 26,734 new pt between 1/1/2005
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PP01.35: MALIGNANT PLEURAL MESOTHELIOMA
IN PATIENTS OLDER THAN 45 YEARS
PP01.36: A MESOTHELIOMA CLUSTER AMONGST
EXPERIMENTAL PHYSICISTS
Fatma M.A. Aou El-Kasem1, Rabab M. Gaafar2, Hala Aziz
Shokralla2, Abdel-Rahman M. Abdel-Rahman3 , Mohamed
Rahoma3
Steven Kazan, William F. Ruiz
Medical Oncology, National Cancer Institute, Cairo,
EGYPT, 2Medical Oncology, NATIONAL CANCER Institute, CAIRO,
EGYPT, 3Surgical Oncology, National Cancer Institute, Cairo,
EGYPT
Kazan McClain Satterley & Greenwood, Oakland, CA, UNITED
STATES OF AMERICA
1
Objectives: A cancer registry was analyzed to determine if the
clinicopathologic characteristics, treatment modalities, and
prognosis of malignant pleural mesothelioma (MPM) patients
>45 years of age at diagnosis differ.
Methods: Retrospective review of patients with MPM presented to medical oncology department, National Cancer Institute,
Cairo University, Egypt;diagnosed between 2007 till 2012. Data
regarding demographics, presentation symptoms, histology,
tumor staging, treatment modality, and survival were obtained
from all patients. Pearson’s chi(2) test and the Kaplan-Meier
method with a log-rank test were used for statistical analysis.
There were 114 cases of MPM diagnosed during this period
2007 and 2012. Among them, 56 patients were > 45 years old .
These patients were selected for our study.
Results: We found that lymph node metastasis (0.047),
progression after initial response (0.009) and WBC (0.002)
significantly affecting PFS of MPM patients older than 45 years.
Median PFS= 8 months. Median OS=17 months. Median age
was 55 years. PS was 0/ 1 in 39 (69%)cases. Thirty four (60%)
cases were males. Fourty-three (76%) cases had asbestosis.
Twenty-seven (48%) cases had chronic disease. Twenty-three
(41%)cases were smokers. Dyspnea was presenting symptom
in 52 (92%)cases. Fourty-nine(87%) cases were complaining
of chest pain. Thirty-five (62%)were complaining of fatigue. Anorexia was present in 14 (25%) cases. Fourty-nine(87%) cases
had effusion. Pleural thickening was documented in 53(94%)
cases. Twenty-six (46%) cases had mediastinal lymph nodes.
Nine (16%) cases had pulmonary metastasis. T1,2,3 represented in 45 (80%) cases. Fifty (89%) cases received platinum containing chemotherapy out of them 42 cases were responders.
Thirty (53%) cases were epithelioid mesothelioma. Twenty-nine
(51%)cases were grade 2&3 of differentiation.
Conclusion: Lymph node metastasis (0.047), progression after
initial response (0.009) and WBC (0.002) significantly affecting
PFS of MPM patients older than 45 years. Early diagnosis and
proper suitable treatment for old MPM patients are recommended. Bigger number of patient is recommended.
Objectives: To determine the cause of mesothelioma in
three renowned physicists who collaborated on experimental
research during the 1960s and 1970s and died of mesothelioma
between 2012 and 2015.
Methods: Eugene Commins, Hyatt Gibbs and Melvin Simmons
each hold doctoral degrees in physics, contributed substantially to their field and received numerous honors. The occupational and asbestos exposure information presented here
was obtained from the public record including available death
certificates,[1] obituaries and information obtained from Mr.
Commins before his death.
Results: Vital Statistics: Eugene Commins was born 1932. He
obtained a Ph.D. in Physics from Columbia University in 1958.
He worked as a physics professor for 57 years. He died in 2015
of Advanced Metastatic Malignant Mesothelioma, Biphasic,
with Bone Metastasis. Hyatt Gibbs was born 1943. He obtained
a Ph.D. in Physics from University of California – Berkeley in
1965. He worked as a research physicist for 43 years. He died
in 2012 of Malignant Pleural Mesothelioma. Melvin Simmons
was born 1943. He obtained a Ph.D. in Physics from University
of California – Berkeley in 1968 and worked as a research physicist thereafter. He died in 2014 of Malignant Mesothelioma.
Occupational Exposure Data[1]:
Professor Commins was a doctoral-level physics professor
at UC Berkeley from 1960-2010. A member of the National
Academy of Sciences and the American Academy of Arts
and Sciences and fellow of the American Association for the
Advancement of Science and the American Physical Society,
his students include Nobel laureate Steven Chu. Dr. Gibbs and
Dr. Simmons were his students. Professor Commins and his
students conducted experimental research that led to important advancements and publications in physics.
During laboratory work during the 1960s and 1970s glass
columns were used. Over time, the columns broke or became
too thin for continued use. Researchers repaired and remade
columns using glass blowing techniques. Asbestos paper and
tape were soaked in water and wrapped around portions of the
glass to allow handling. When the repairs were finished, the
dried asbestos products were scraped into a trash can. Tongs
with asbestos sleeves and asbestos gloves were used to handle
hot glass and other high temperature materials. During circuit
board soldering, transistors and other components were covered with torn-off asbestos tape to insulate the components. Conclusion: The subjects here were exposed to the same
asbestos products in the same laboratory during the same
time and died of mesothelioma within a three year period, five
decades later. This data supports a prior mesothelioma cluster
in an occupation not traditionally associated with asbestos
exposure, and serves as a reminder that inhalation of asbestos
fibers, not job title, increases mesothelioma risk. [1] Dr. Gibbs
died in France; his death certificate was not obtained.
[2] Professor Commins reported that he cut asbestos ceiling tile
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for two weeks in the 1950s, and repaired drywall at his home on
two occasions in the 1960-70s. We found no information that
Dr. Gibbs or Dr. Simmons had other occupational or non-occupational exposures to asbestos.
with good PS. Keywords: mesothelioma, Physicists, laboratory asbestos
exposure, cancer cluster
PP01.37: SURVIVAL IN GOOD PERFORMANCE
MALIGNANT PLEURAL MESOTHELIOMA PATIENTS;
PROGNOSTIC FACTORS AND PREDICTORS OF
RESPONSE
M Rahouma1, Hala Aziz Shokralla2, Rabab M. Gaafar2, Galal
Ghaly1, Abdelrahman Mohamed1
Surgical Oncology, National Cancer Institute, Cairo,
EGYPT, 2Medical Oncology, National Cancer Institute, Cairo,
EGYPT
1
Keywords: Malignant pleural mesothelioma (MPM), performance status, prognostic factors and predictors of response to
chemotherapy
Objectives: Malignant pleural mesothelioma (MPM) has a poor
prognosis in general, we sought to evaluate prognostic factors
and predictors of response to chemotherapy in good ECOG-performance status (PS=0-1) patients.
Methods: We retrospectively reviewed our database and
enrolled 82 patients with histologically confirmed MPM and
PS=0-1 between 2012-2014 Age, weight, gender, smoking
status, comorbidities, asbestosis, different symptoms, Tumor,
Nodal(N)and Metastasis stages, response, different laboratory
values, including pretreatment haemoglobin(Hb), neutrophil/
lymphocyte ratio and pathology. Survival was analyzed using
Cox regression in univariate and multivariate analysis. Kaplan–Meier survival curves were obtained and compared by
log–rank. Logistic regression was used to determine factors
predicting response
Results: Eligible patients were 82(Median age 45years, median
body Weight 77 Kg, Hb=12 g/dl, platelets =372 x 109 /L , leukocyte=9.7x 103 /µL, neutrophils=6.1 cells/µL, lymphocytes=1.89
cells/µL, neutrophils-lymphocytes ratio (NLR)=3.6 pretreatment). Forty three were men, thirty cases were smoker, and 65
had asbestosis. twenty three present with chronic disease. all
cases received platinum based chemotherapy;55(67.07%) were
responder(whether SD or PR). Pathology were 49(59.8%) epithelial type, 17(20.7%) mixed type and 16(19.5%)sarcomatoid
type. Median overall survival were 17 months (95%CI=14.1119.90). Median progression free survival(PFS) were 9
months(95%CI=6.97-11.03). Significant decrease in PFS were
observed among advanced nodal (N) disease (median PFS in
N0 and N+ were 10 and 5 months respectively), non-responders(p=0.012), lower NLR(p=0.026) and epithelial pathology
type (p=0.062). Multivariate analysis(MVA) demonstrated that
advanced N status (p=0.015), non-responder (p<0.001), lower
NLR (p=0.015) and smoking (p=0.07) adversely affecting prognosis. Multivariate analysis shows only absence of metastasis(M0) (p=0.04) were the significant predictor of response
Conclusion: In addition to previously recognized prognostic
factors in MPM, better median survival is evident in patients
PP01.38: NON-EPITHELIAL PLEURAL
MESOTHELIOMA; CRITERIA, PROGNOSTIC
FACTORS AND PREDICTORS OF RESPONSE
Hala Aziz Shokralla1, M Rahouma2, Iman Loay3 , Rabab M.
Gaafar1, Abdelrahman Mohamed2
Medical Oncology, National Cancer Institute, Cairo, EGYPT, 2Surgical Oncology, National Cancer Institute, Cairo, EGYPT, 3Cancer
Pathology Depatement, National Cancer Institute, Cairo, EGYPT
1
Objectives: Malignant pleural mesothelioma (MPM) has a
poor prognosis in general. We conducted this study to evaluate
prognostic factors and predictors of response to chemotherapy
in non-epithelial MPM.
Methods: We retrospectively reviewed our database and
included patients with histologically confirmed non epithelial
MPM 2012-2014. Age, weight, gender, smoking status, performance status(PS), comorbidities, asbestosis, symptoms, Tumor,
Nodal(N)and Metastasis stages, response, laboratory values,
including pretreatment haemoglobin(Hb), neutrophils/lymphocytes ratio(NLR), and pathology. Survival was analyzed using
Cox regression in univariate and multivariate analysis. Logistic
regression was used to determine factors predicting response.
Kaplan–Meier(KM) survival curves were obtained and compared by log–rank.
Results: Enrolled patients were 47(Median age 48years, Weight
80Kg, Hb=12 g/dl, platelets=343x109 /L, leukocytes=11x103 /
µL, neutrophils=6.1 cells/µL, lymphocytes=1.10 cells/µL,
NLR=5.25 pretreatment). Twenty eight were men from which
21 were smokers, 40 had asbestosis. Thirteen cases had
chronic disease, 33(70.2%) were responders. Pathology were
25(53.2%) sarcomatoid type and 22(46.8%) mixed. Median
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overall survival(OS) were 17 months(95%CI=13.70-20.30).
Median OS in N0(65.96%) versus N+(34.04%) were 19 months
versus 9 months respectively(p=0.033) (figure 1). Median progression free survival(PFS) were 8 months(95%CI=5.61-10.39).
Multivariate analysis(MVA) model included variables that had
p value <0.20 on univariate analysis and involved weight,
gender, smoking status, response and lymphocytes and revealed that high body weight [p=0.034(Hazard ratio(HR)=1.03,
95%CI=1.01-1.05)]and absence of response to chemotherapy(progressive disease) [(p=0.048 (HR=0.048,CI=0.24-0.99)]
adversely affecting prognosis. There were no difference in
OS(p=764) or PFS(p=0.676) between sarcomatoid and mixed
pathology using unadjusted KM curves. Predictors of response,
using Logistic regression, revealed that presence of asbestosis(
p=0.038) was the only significant predictors of poor response to
chemotherapy.
Keywords: prognostic factors and predictors of response to
chemotherapy, Asbestosis, Non-epithelial malignant pleural
mesothelioma (MPM), Body weight
PP01.39: DOES PRETREATMENT BODY WEIGHT
HAVE ANY SIGNIFICANT IMPACT ON SURVIVAL IN
PLEURAL MESOTHELIOMA IN YOUNG AGE GROUP?
M Rahouma1, Hala Aziz Shokralla2, Rabab M. Gaafar2,
Mohamed Kamel1, Abdelrahman Mohamed1
Surgical Oncology, National Cancer Institute, Cairo,
EGYPT, 2Medical Oncology, National Cancer Institute, Cairo,
EGYPT
1
Conclusion: Better OS in non-epithelial mesothelioma type
was observed among N0 stage reflecting importance of early
detection. Worse PFS was observed among those with heavy
body weight suggesting disadvantages of overeating. Asbestosis
carries a poor prediction to chemotherapy among our cohort. Multivariate analysis of PFS and MVA for predictors of
response MVA of PFS
P value
HR
95% CI
(Upper)
95% CI
(Lower)
Weight
0.034
1.03
1.01
1.05
Gender
0.154
2.01
0.77
5.27
Smoking status
0.369
1.56
0.59
4.12
Response
0.048
0.49
0.24
0.99
Lymphocytes
0.070
1.51
0.97
2.36
MVA for predictors of response
P value
OR
95% CI
(Upper)
95% CI
(Lower)
PS
0.091
0.011
0.01
1.43
Asbestosis
0.038
0.05
0.01
0.85
Chest pain
0.080
0.09
0.01
1.35
Leukocytes
0.269
0.85
0.64
1.13
Objectives: Nutritional status has been associated with long
term outcomes in cancer patients. We investigated whether
the body weight (BW), as an indicator of the nutritional status,
affects the survival in malignant pleural mesothelioma (MPM)
patients in young age group.
Methods: We retrospectively reviewed our database and enrolled patients with histologically confirmed MPM and aged 45
years or below in the period between 2012-2014 who received
chemotherapy. Age, weight, gender, smoking status, presence
of chronic diseases, asbestosis, different symptoms, Tumor(T),
Nodal(N) and Metastasis(M) stages, response, different
laboratory values, including pretreatment hemoglobin(Hb),
neutrophils/lymphocytes ratio(NLR), and pathology. Survival
was analyzed using Cox regression in univariate and multivariate analysis. Kaplan–Meier survival curves were obtained and
compared by log–rank.
Results: Eligible patients were 58 (Median age 39 years, median body Weight 77.5 Kg, pretreatment haemoglobin 12 g/dl,
platelets 367 x 109 /L, leukocytes 10.7 x 103 /µL, neutrophils 6
cells/µL, lymphocytes1.25 cells/µL, NLR 4.5). Thirty-two cases
(55.2%) were men, nineteen were smokers, 43 had asbestosis
and only six cases presented with chronic disease. All cases
received platinum based treatment; 17 cases (29.31%) were
responder (whether stable disease or partial responders).
Pathology were 37 (63.8%) epithelial type, 13(22.4%) were
sarcomatoid type and only eight (13.8%) were mixed type.
Median overall survival were 16 months (95% Confidence
Interval (CI)=13.55-18.45). Median progression free survival
(PFS) were 8 months(95%CI=6.17-9.84). PFS were adversely
affected by Weight, as a continuous variable [p=0.025, Hazard
Ratio(HR)1.02, CI:1.01-1.04] and advanced T stage (see figure).
Conclusion: Although BW may reflects the nutritional status,
that has been associated with long term outcomes in cancer
patients, but it is associated with poor PFS in MPM.
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was applied. In 56 patients (54%) chemotherapy was applied
(most common chemotherapy regiemens were pemetrexedcisplatin and gemcitabin-cisplatin). Radiotherapy was applied
in 10 patients. Median overall survival (OS) was in all group of
patients 11 months. Better OS was observed in epitheliod type
compared to sarcomatoid type (11 vs 5,5 months). In patients
with mixed type mean OS was 12,5 months. OS was significantly influenced by stage. OS was significantly better in stage III
compared with stage IV (17,5 vs 9 months).
Conclusion: Our data showed that surgery and radiotherapy
are not preferd choice of treatment in our institution. Majority of
patient were treated with chemotherapy alone. OS of our data
showed comparable results with previously published data.
Showing improvent in OS in stage III compared with stage IV
patients. The study emphase need for earlier diganosis of MPM
patients which could lead to better outcome in MPM patients.
Keywords: prognostic factors and predictors of response to
chemotherapy, Malignant pleural mesothelioma (MPM), Body
weight
PP01.40: MALIGNANT PLEURAL MESOTHELIOM: A
SINGLE-CENTER EXPERIENCE IN CROATIA
Marko Jakopovic1, Luka Brcic2, Tomislav Dujmovic3 , Goran
Glodic3 , Anton Mazuranic3 , Branka Cucevic4 , Suzana Kukulj4 ,
Sanja Plestina4 , Mihovil Roglic4 , Zoran Janevski4 , Ivica
Mazuranic4 , Silvana Smojver-Jezek4 , Sven Seiwerth3 ,
Miroslav Samarzija4
Department For Respiratory Diseases Jordanovac, University
Hospital Center Zagreb, Zagreb, CROATIA, 2University of Graz,
Graz, AUSTRIA, 3Zagreb Medical School, Zagreb, CROATIA, 4University Hospital Centre Zagreb, Zagreb, CROATIA
1
Objectives: Malignant pleural mesothelioma (MPM) is a
rare malignancy usually caused by asbestos exposure. In this
retrospective study, we aimed to analyse demographic, clinical,
and pathological data and treatment-related outcomes in MPM
patients diagnosed and treated in our institution.
Methods: In this retrospective analysis we included 104 MPM
patients diagnosed at Department for Respiratory Diseases
Jordanovac, University Hospital Center Zagreb, Croatia.
Results: Between years 1999 and 2012, 104 patients were
diagnosed with MPM (91 males, 13 females, mean age at diagnosis was 62). Epitehelial type was present in 74 patients (71%),
sarcomatoid in 5 patients (4,8%), mixed type in 1 patient (1%),
and in 21 (20%) the subype could not be determend. The disase
was stage as IV in 70 patients (67%), stage III in 27 patients
(26%), and as stage II in 4 patients (3%). The most frequent
metastatic sites were mediastinum, thoracic wall, diaphragm
and lungs. Out of 104 patients only 15 patients (14%) were
surgically treated. Surgical procedures included decortication and pleurectomy. No extra-pulmonary pleurectomy was
performed. In 36 patients (35%) onyl best supportive care
Keywords: malignant pleural mesothelioma, overall survival,
chemotherapy
PP01.41: MESOTHELIOMA INCIDENCE IN
LOMBARDY, ITALY: TIME PATTERNS AND FUTURE
PROJECTIONS
Dario Consonni1, Sara De Matteis2, Barbara Dallari1, Luciano
Riboldi1, Pier Alberto Bertazzi1, Carolina Mensi1
Department of Preventive Medicine, Fondazione IRCCS Ca’
Granda - Ospedale Maggiore Policlinico, Milan, ITALY, 2National
Heart & Lung Institute, Occupational & Environmental Medicine,
Imperial College London, London, UNITED KINGDOM
1
Objectives: Measuring malignant mesothelioma (MM) or
pleural cancer incidence/mortality is a useful means to monitor
asbestos-related diseases occurrence and to identify sources
of asbestos exposure. Using data of the MM registry of the
Lombardy Region, North-West Italy, the most populated and
industrialised Italian region, we analysed asbestos exposure
and time patterns in the period 2000-2012 and made future
projections for the period 2013-2029.
Methods: We selected all incident cases of MM with first
diagnosis between 2000 and 2012. We examined time trends
using standardised rates and Poisson regression. We fitted
categorical Poisson age-cohort models using 5-year categories
for age at diagnosis (reference: 70-74 years) and birth cohort
(reference: cohort 1920-1924). The gender-specific age and
cohort regression coefficients were then applied to population
data to calculate projections of the numbers of MM cases and
their 90% confidence intervals (CI) in the years 2013 to 2029.
Data management and statistical analyses were performed with
Stata 13.
Results: In 2000-2012 we recorded 4,435 MM cases, 2,846 in
men and 1,589 in women. Occupational asbestos exposure was
more frequent in men (73.6%) than in women (38.2%). The average number of MM cases per year was still increasing (+2.6%
in men, +3.3% in women). A maximum of 416 MM cases (266
men, 150 women) is expected in 2019. We forecast there will be
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6,809 more cases (4,379 in men, 2,430 in women) in the period
2013-2029, for a total of 11,244 MM cases (7,225 in men, 4,019
in women) in 30 years.
Conclusion: This study documented a high MM burden in
both genders in the Lombardy Region, reflecting extensive
occupational (mainly in men) and non-occupational (mainly in
women) asbestos exposure in the past. Incidence rates are still
increasing and a downturn of MM occurrence is expected to
occur after 2019. Documenting mesothelioma occurrence may
help to increase awareness of dangers of asbestos exposure
in countries that still use it but where its health effects are still
overlooked.
Keywords: mesothelioma projections, mesothelioma incidence, asbestos, mesothelioma registry
patients are recommended.
Keywords: mesothelioma, young, clinicopathological,PFS
PP01.43: INCREASING AGE AT DIAGNOSIS
IN THE AUSTRALIAN MALIGNANT PLEURAL
MESOTHELIOMA POPULATION: WHAT ARE THE
POTENTIAL IMPLICATIONS?
Matthew J. Soeberg1, James Leigh1, Tim Driscoll2, Bruce Armstrong2, Jane Young2, Nico Van Zandwijk1
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA, 2Sydney School of Public Health, The University of Sydney,
Sydney, NSW, AUSTRALIA
1
PP01.42: CLINICO PATHOLOGICAL
CHARACTERISTICS OF YOUNG EGYPTIAN
MALIGNANT PLEURAL MESOTHELIOMA PATIENTS
Fatma M.A. Aou El-Kasem1, Abdel-Rahman M. Abdel-Rahman2,
Mohamed Rahoma2
Medical Oncology, National Cancer Institute, Cairo, EGYPT, 2Surgical Oncology, National Cancer Institute, Cairo, EGYPT
1
Objectives: A cancer registry was analyzed to determine if the
clinicopathologic characteristics, treatment modalities, and
prognosis of malignant pleural mesothelioma (MPM) patients <
45 years of age at diagnosis differ. Tables ,pictures, images are
not applicable
Methods: There were 114 cases of MPM diagnosed during
this period 2007 and 2012. Among them, 58 patients were <
45 years old . These patients were selected for our study. Data
regarding demographics, presentation symptoms, histology,
tumor staging, treatment modality, and survival were obtained
from all patients. Pearson’s chi(2) test and the Kaplan-Meier
method with a log-rank test were used for statistical analysis
Results: We found that weight , T4 and neutrophil lymphocytes
ratio significantly affecting PFS of MPM patients younger than
45 years. Median PFS= 8 months. Median OS=16 months. Our
patients’ age ranged from 19 to 45 years. PS was 0/ 1 in 43
cases. Thirty two cases were males. Six cases had Asbestosis. Nineteen cases were smokers. Dyspnea was symptom in
53 cases. Fourty-nine cases were complaining of chest pain
,Fourty-one were complaining of fatigue, anorexia was present
in 21 cases, fifty one cases had effusion, Pleural thickening was
documented in 57 cases, Twenty-four cases had mediastinal
lymph nodes. Three cases had pulmonary metastasis, T1,2,3
represented in 50 cases. Fifty-two cases received platinum containing chemotherapy out of them 17 cases were responders.
Thirty-seven cases were epithelioid mesothelioma, thirty-two
cases were grade 2&3 of differentiation
Objectives: Australia is known to have had one of the highest
per-capita asbestos consumption rates, yet there are few contemporary reports on malignant mesothelioma trends.
Methods: Data on 10,930 people with malignant pleural mesothelioma and 640 people with malignant peritoneal mesothelioma diagnosed in Australia during 1982-2009 were analysed.
Observed incidence rate trends were quantified. Using age-period-cohort analyses, age-specific incidence rates were projected
up to 2030 using observed incident cases during 1982-2012.
Results: During 1982-2009, acceleration in malignant pleural
mesothelioma age-standardised incidence rates were highest
for women and those aged 75 years and above, with average
annual percentage changes of +4.9 (95% CI 3.6, 6.2) and +7.2
(95% CI 5.4, 9.0) respectively. Age-standardised incidence
rates for men with malignant pleural mesothelioma aged 0-64
years decelerated rapidly during 2003-2009, an average annual
percentage change of -5.1% (95% CI -7.6, -2.5). Overall, male
age-specific malignant pleural mesothelioma incidence rates in
the 65-74 year age group during 2010-2030 are projected to decline with rates projected to increase for older men and women
with malignant pleural mesothelioma.
Conclusion: In Australia, there is a marked increase over time
in people aged 75 years or more diagnosed with malignant
pleural mesothelioma. In this presentation, we explore Australian and international data on malignant pleural mesothelioma
case-series to investigate the potential treatment implications
of this shift over time in malignant pleural mesothelioma age
group distributions.
Keywords: Australia; malignant pleural mesothelioma; incidence; trends
Conclusion: Weight , T4 and neutrophil lymphocytes ratio
significantly affecting PFS of MPM patients younger than 45
years.early diagnosis and agreesive treatment for young MPM
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PP01.44: STUDY ON THE EVOLUTION OVER TIME
OF THE RISK OF MESOTHELIOMA AND LUNG
CANCER AMONG FORMER ASBESTOS-EXPOSED
WORKERS
Corrado Magnani1, Laura Ancona2, Antonio Baldassarre3 ,
Vittoria Bressan4 , Tiziana Cena1, Elisabetta Chellini5, Francesco
Cuccaro6 , Daniela Ferrante1, Patrizia Legittimo7, Ferdinando Luberto8 , Alessandro Marinaccio9, Stefano Mattioli7, Simona Menegozzo10, Enzo Merler4 , Lucia Miligi5, Dario Mirabelli11, Marina
Musti3 , Enrico Oddone12, Venere Pavone13 , Patrizia Perticaroli14 ,
Aldo Pettinari14 , Roberta Pirastu15, Alessandra Ranucci1, Elisa
Romeo2, Orietta Sala16 , Corrado Scarnato13 , Stefano Silvestri17
Dep. Translational Medicine, University Eastern Medicine, Novara, ITALY, 2Department of Epidemiology, Lazio Regional Health
Service, Rome, ITALY, 3Interdisciplinary Department of Medicine,
Occupational Medicine “B. Ramazzini”, University of Bari, Bari,
ITALY, 4Mesothelioma Register of the Veneto Region, Padua Local
Health Unit, Padova, ITALY, 5Occupational & Environmental
Epidemiology Unit-Cancer Research & Prevention Institute (ISPO),
Florence, ITALY, 6Azienda Sanitaria Locale BAT (Barletta, Andria,
Trani), Unità Operativa Epidemiologia e Statistica, Barletta,
ITALY,7Department Medical and Surgical Sciences, University of
Bologna, and Unit of Occupational Medicine, S.Orsola-Malpighi
University Hospital, Bologna, ITALY, 8Inter-institutional Epidemiology Unit, AUSL Reggio Emilia and Arcispedale Santa Maria Nuova, IRCCS, Reggio Emilia, ITALY, 9Italian Workers’ Compensation
Authority (INAIL), Department of Occupational and Environmental Medicine, Epidemiology and Hygiene, Unit of Occupational
and Environmental Epidemiology, Italian Mesothelioma Register,
Rome, ITALY, 10National Cancer Institute IRCCS Fondazione
Pascale, Napoli, ITALY, 11Unit of Cancer Epidemiology, CPO
Piemonte and University of Turin, Torino, ITALY, 12Department of
Public Health, Experimental and Forensic Medicine, University of
Pavia, Pavia, ITALY, 13Epidemiology Unit, Local Health Authority,
Bologna, ITALY, 14Prevention Department, ASUR Marche, Senigallia, ITALY, 15Department of Biology and Biotechnologies Charles
Darwin, Sapienza Rome University, Rome, ITALY, 16A.R.P.A. Emilia
Romagna, Sezione Provinciale di Reggio Emilia, Reggio Emilia,
ITALY, 17Cancer Prevention and Research Institute (ISPO), Firenze,
ITALY
1
Objectives: According to the IARC, asbestos is carcinogenic
to humans, and exposure to asbestos results in mesothelioma,
lung cancer, ovarian and laryngeal cancer with sufficient evidence. It may also lead to gastric, colorectal or pharyngeal cancer with more limited evidence. The possible reduction in risk
after cessation of exposure and after time since first exposure
more than 40 years is still matter of debate. The cessation of
the use of asbestos in Italy in 1992 has lead to a situation similar to a natural experiment that measures large-scale trends in
the risk of disease among former asbestos-exposed workers.
Methods: The study included a pool of Italian cohorts of
asbestos exposed employed in plants located in different Italian
regions that have already been the subject of epidemiological study. The follow up was updated up to 2010 or later. The
main production sectors are: asbestos cement, construction
and maintenance of rolling stock, shipbuilding. We computed
SMRs for the major causes of death. The number of deaths
expected in the cohort was estimated from age and sex-specific
mortality rates of the participating Italian regions, provided by
ISTAT (Rome, Italy) for the period 1970-2012. The incidence of
mesothelioma will be detected using a record linkage to the
National Mesothelioma Registry (ReNaM). A pooled analysis is
performed in order to obtain information on the variation in the
risk of malignant mesothelioma and the risk of death from other
malignancies. In order to obtain a quantitative estimation of
the exposure level to asbestos of the subjects of the cohorts, an
index that takes into account the fraction of exposed to asbestos, direct or indirect use of asbestos and exposure level was
computed. Information about these aspects has been provided
for each cohort.
Results: Pooled cohorts study includes 54,409 subjects, of
which 14,743 have worked in the production of cement - asbestos. The dataset includes 48,355 men and 6,054 women. At the
end of follow-up, 55.3% of the subjects were alive, 43.0% had
died, and 1.7% were lost to follow-up or had moved abroad. The
cause of death was known for the 94% of deceased subjects.
Considering the follow up period after 1970, the cohorts contributed altogether about 1,430,000 person-years for men and
184,000 for women. Both genders showed increased mortality
for all causes (p<0.001), all malignancies (p<0.01), pleural
and peritoneal malignancies (both p<0.01) and lung cancer
(p<0.01). In women, ovarian malignancies were more frequent
than expected (p<0.05). No statistically significant increase was
found for laryngeal cancer.
Conclusion: In addition to the main objective of assessment
of risk over time, it is expected that the study will allow further
results, such as assessing the risk for those malignancies still
under discussion and studying the risk for women. The working
group: M.N.Ballarin4 , F.Barone-Adesi18 , C. Brentisci11, B.Cortini5, S.Curti7, M.Gangemi11, P.Girardi4 , F.Gioffrè4 , G.Gorini5,
L.Mangone8 , F.Marinelli7, P.Marinilli13 , C.Panato4 , F.Roncaglia8 ,
C.Storchi8 , A.Stura11, S.Tunesi1, M.Vicentini8 , S.Verdi5, A.M.
Nannavecchia19, L.Bisceglia20 1-17 as Authors’ affiliations 18Department of Pharmaceutical Sciences, University of Eastern
Piedmont, Novara, Italy 19IRCCS Giovanni XXIII Oncologico Bari,
Puglia, Italy 20 Agenzia Regionale Sanità Puglia, Bari, Italy
Keywords: asbestos, mesothelioma, latency
PP01.46: MORTALITY/HOSPITALIZATION FROM
PLEURAL MESOTHELIOMA ASSOCIATED WITH
ENVIRONMENTAL EXPOSURE TO FLUOROEDENITE IN BIANCAVILLA
Susanna Conti1, Valerio Manno1, Giada Minelli1, Caterina Bruno2, Lucia Fazzo3 , Pietro Comba3
Unit Of Statistics, Istituto Superiore di Sanità, Rome, ITALY, 2Environment And Prevention, Istituto Superiore di Sanità, Roma, ITALY, 3Environment And Prevention Department, Istituto Superiore
di Sanità, Roma, ITALY
1
Objectives: Fluoro edenite, a fibrous amphibole present in
the sole and building materials of Biancavilla, a small town in
Sicily (ITALY) was allocated in 2014 by the International Agency
for Research on Cancer (IARC) to the Group 1 (“the agent is
carcinogenic to humans”) as cause of mesothelioma. This evaluation was based on epidemiological studies coordinated by
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ABSTRACT BOOK
Istituto Superiore di Sanità. The aim of our research is to study
mortality and hospitalization from mesothelioma among residents in Biancavilla, using current data at the best of updating
in Italy, available at the Unit of Statistics of Istituto Superiore di
Sanità.
Methods: The most recent national official mortality and hospitalization data (2006-2012) were analyzed. Mortality data are
codified according to the International Classification of Disease
(ICD), 10th Revision, which, introducing the morphology of malignant neoplasms, has specific codes for Pleural and Peritoneal
Mesothelioma. Hospitalization data are yet codified according
to the ICD Clinical Manifestation 9th Revision, which has only
a topographic classification of neoplasms, therefore, hospitalizations due to malignant neoplasm of pleura/peritoneum were
investigated. Standardized Mortality Ratio (SMR), Standardized
Hospitalization Ratio (SHR) and their 90%Confidence Intervals
(CI) were estimated by gender and age, being the population
of Sicily the reference. To control confounding from social and
economic factors, the SMRs and SHRs were adjusted by a
deprivation index.
Results: Among residents in Biancavilla there was a statistically significant death excess from Pleural Mesothelioma: SMR
632 (CI 409-978), based on 14 deaths during the study period,
both among men and women. When SMR estimates were stratified by age, very high figures were shown in the younger age
groups: SMR 1741, CI 576-5262 in subjects less than 50 years
old and SMR 3229, CI 720-14474 in subjects less than 40 years
old. The sex ratio (M/W) of deaths from pleural mesothelioma
was 0.75 overall, 0.50 among younger subjects. Also hospitalizations due to malignant pleural cancer (18 cases) showed
excesses, overall (SHR 399, CI 271-587) and in particular
among younger persons (aged less than 50 years): SHR 1078,
CI: 484-2400; the sex ratio was 1.25. Neither deaths nor hospitalizations from peritoneal mesothelioma were observed.
Conclusion: The observed sex ratio (close/less than one) corroborates the hypothesis of an environmental exposure, rather
than occupational; moreover, this may also reflect a high level
of fiber exposure for women, who are often engaged in activities
such as sweeping of floors, balconies and sidewalks located in
front of the houses. The latency period of Pleural Mesothelioma
caused by exposure to fluoro-edenitic fibres has not yet been
studied, but if it were similar to the latency of Pleural Mesothelioma due to asbestos (20 - 40 years) the higher excesses
of deaths and hospitalizations that we observed among young
subjects would suggest early exposure, in teenage/childhood
years. Major clean-up interventions were performed in Biancavilla after its recognition as a National Priority Contaminated
site (2002) but our results suggest that particular attention
should be paid on community needs in term of early diagnostic
procedure and medical care, for both genders.
PP01.47: SURVIVAL AND EXPECTED YEARS OF
LIFE LOST OF MALIGNANT MESOTHELIOMA:
ANALYSIS OF 105 CASES IN TAIWAN, 1977-2015
Lukas J. Lee1, Ting-Hui Wu2, Yu-Yin Chang1, Jung-Der Wang3
National Institute of Environmental Health Sciences, National
Health Research Institutes, Zhunan Town, TAIWAN, 2Department
of Oncology, National Taiwan University Hospital, Taipei, TAIWAN, 3Department of Public Health, College Of Medicine, National Cheng Kung University, Tainan, Taiwan, College of Medicine,
National Cheng Kung University, Tainan, TAIWAN
1
Objectives: Malignant mesothelioma (MM) is a rare cancer
with limited information on survival. We investigated clinical
factors associated with survival and estimated life years lost
based on a patient series in Taiwan.
Methods: We retrospectively reviewed medical records of
patients diagnosed with MM at the National Taiwan University Hospital from 1977 to August 2015, with follow-up of
vital status through 31 October 2015. Assuming a constant
excess hazard, we extrapolated lifetime survival function by a
semi-parametric method. For each MM patient, we simulated
age- and gender-matched referents based on the vital statistics of Taiwan to estimate expected years of life lost (EYLL) as
an indicator of health gap. For recognizing prognostic factors
associated with pleural MM, we performed univariate analyses
using Kaplan–Meier survival functions with log-rank tests. Then
multivariate Cox regression models were performed to identify
significant risk predictors.
Results: A total of 105 cases of MM were included. The mean
age at diagnosis was 56.7±14.0 years. The EYLL due to MM was
20.2 years. There were 82 pleural MM, 17 peritoneal MM, 4 diffuse MM, and 2 testicular MM. The overall median survival for
pleural MM was 14.8 months. Cox regression models revealed
that age less than 65 years, clinical stages I~II, ECOG (Eastern
Cooperative Oncology Group) performance status 0-1, and
surgical operation were associated with longer survival.
Conclusion: Substantial life years lost resulted from MM was
found. Age less than 65, early stages, good performance status,
and surgical operation were independent prognostic factors of
pleural MM.
Keywords: prognostic factor, Malignant mesothelioma, expected years of life lost
Keywords: Fluoro-edenite, mortality/hospitalization, early
exposure, pleural mesothelioma
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ABSTRACT BOOK
PP01.50: INVESTIGATING PALYGORSKITE’S ROLE
IN THE DEVELOPMENT OF MESOTHELIOMA IN
SOUTHERN NEVADA
David Larson1, Amy Powers1, Jean-Paul Ambrosi2, Mika Tanji1,
Andrea Napolitano1, Erin Flores1, Francine Baumann1, Laura
Pellegrini1, Cormac Jennings1, Brenda Buck3 , Brett Mclaurin4 ,
Doug Merkler5, Cleo Robinson6 , Paul Morris1, Meral Dogan7,
A. Umran Dogan7, Harvey I. Pass8 , Sandra Pastorino1, Michele
Carbone1, Haining Yang1
STATES OF AMERICA, 2Aix-Marseille Université, Provence,
FRANCE, 3University of Nevada Las Vegas, Las Vegas, NV,
UNITED STATES OF AMERICA, 4Bloomsburg University of Pennsylvania, Bloomsburg, AL, UNITED STATES OF AMERICA, 5USDA,
Natural Resources Conservation Service, Las Vegas, NV, UNITED
STATES OF AMERICA, 6University of Western Australia, Harry
Perkins Institute for Medical Research, Nedlands, Perth, ACT,
AUSTRALIA, 7University of Iowa, Iowa City, IA, UNITED STATES
OF AMERICA, 8New York Langone Medical Center, New York, NY,
1
Objectives: Similar to asbestos fibers, non-regulated mineral
fibers can cause malignant mesothelioma (MM). Recently,
increased proportions of women and young individuals with
MM were identified in southern Nevada, suggesting that
environmental exposure to carcinogenic fibers was causing the
development of MM. Palygorskite, a fibrous silicate mineral with
a history of possible carcinogenicity, is abundant in southern
Nevada. In this study, our aim was to determine whether palygorskite was contributing to the development of MM in southern
Nevada.
PP01.51: MESOTHELIOMA MORTALITY IN POLAND
BETWEEN 1999 AND 2013
Gabriela Oledzka1, Ewa Wilk2, Agnieszka Skubiszewska1, Anna
Minkiewicz1, Małgorzata Krowczynska2
Department of Medical Biology, Medical University of Warsaw,
Warsaw, POLAND, 2Department of Geoinformatics And Remote
Sensing, University of Warsaw, Warsaw, POLAND
1
Objectives: Many Western countries are currently suffering
from a malignant pleural mesothelioma (MM) the increasing number of cases. Poland belongs to countries with low
incidence rates or insufficient data of morbidity and mortality.
In Poland, after asbestos was banned in 1997, the problems
caused by asbestos focused on monitoring the health of workers with prior exposure to this substance and those currently
involved in the demolition of asbestos-containing buildings and
in asbestos removal tasks. In Poland as in various countries of
Central-Eastern Europe, the crude incidence of mesothelioma
appeared to be lower than in Western countries. The aim of this
study was to evaluate the variations of pleural MM incidence in
Poland.
Methods: This article is based on a selective review of the
literature, along with data from the central database of the National Cancer Registry. The evolution of pleural mesothelioma
between 1999 and 2013 was investigated using data collected
in 16 voivodship.
Results: Methods: We studied and compared the toxicity and carcinogenesis of palygorskite fibers vs. crocidolite asbestos using our
established in vitro and in vivo systems.
Results: While palygorskite, in vitro, displayed some cytotoxicity towards HM cells and reduced their viability, the effects were
roughly half of those observed when using similar amounts of
crocidolite asbestos. No Balb/c (0/19) or MexTAg (0/18) mice
injected with palygorskite developed MM, while 3/16 Balb/c and
13/14 MexTAg mice injected with crocidolite did. Lack of MM
development was associated with a decreased acute inflammatory response, as injection of palygorskite resulted in lower
percentages of macrophages (p=0.03) and neutrophils (p=0.02)
in the peritoneal cavity 3 days after exposure. Additionally, compared to mice injected with crocidolite, palygorskite-injected
mice had lower percentages of M2 (tumor-promoting) macrophages (p=0.008) in their peritoneal cavities when exposed to
fiber for several weeks.
The evolution of the MM incidence rates in men and women
between 1999 and 2013 in Poland are described in Fig 1. The
incidence rate for 100.000 person-year in men was 0.18 in 1999
and was estimated to be 0.65 in 2013. The incidence rate for
100.000 person-year in women increased between 0.06 in 1999
and 0.23 in 2013. Conclusion: Our study indicates that palygorskite found in the
environment in southern Nevada does not cause MM in mice,
seemingly because palygorskite, in vivo, fails to elicit inflammation that is associated with MM development. Therefore, palygorskite is not a likely contributor to the MM cases observed in
southern Nevada.
Keywords: Palygorskite, environment, mesothelioma, Nevada
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ABSTRACT BOOK
lioma for men and women in 2013. Very few MMs occurred
before the age of 45 years, and, in both genders, MM steadily
increased at each subsequent age, up to age 60.
Conclusion: The incidence of mesothelioma has increased
off since 2012. Our results suggest that in Poland the national
incidence of mesothelioma is not expected to drop in the next
few years. Acknowledgements: This study was supported by the
Ministry of Economy, Programme for Asbestos Abatement in
Poland 2009–2032.
Keywords: asbestos; malignant pleural mesothelioma, incidence
PP01.52: RELATIONSHIP BETWEEN
HISTOLOGICAL TYPE AND PLEURAL EFFUSION IN
Taiichiro Otsuki, Koji Mikami, Takayuki Terada, Kozo Kuribayashi, Takashi Nakano
Internal Medicine, Hyogo College of Medicine, Nishinomiya,
JAPAN
Objectives: Pleural effusion is important for the diagnosis
and treatment of malignant pleural mesothelioma (MPM), but
an analysis of the clinical pathology is not always performed.
The cytology positive rate for pleural effusion in MPM is lower
than that in lung cancer (approximately 25–30%). This study
analyzed the relationship between histological type and pleural
effusion for elucidating the clinical findings of the latter in MPM.
Methods: This study is a retrospective analysis of 143 patients
(115 males and 28 females) who received a pathological diagnosis of MPM in our hospital from 2011 to 2014. Parameters
analyzed include the existence of pleural effusion upon the
patient’s initial visit, histological type , and clinical staging.
Results: Histological type was epithelioid in 106 cases, biphasic in 15, and sarcomatoid in 22. There were 14 cases with
no pleural effusion (epithelioid in four cases, biphasic in two,
and sarcomatoid in eight). These cases were trending to the
progression stage (IMIG stageⅠ/Ⅱ/Ⅲ/Ⅳ =1/1/2/9). The cytology
positive rate for effusion in the epithelioid-type MPM was higher
than in the other types of MPM (epithelioid/biphasic/sarcomatoid=34%/8.0%/7.1%).
Conclusion: It has been theorized that many mesothelioma
cells in pleural effusion are cells peeling from the epithelioid
type and the epithelioid type component in the biphasic type.
Therefore, it is suggested that the cytology positive rate for effusion in epithelioid-type MPM is higher than in the other types.
One of the reasons for having fewer cases with pleural effusion
in sarcomatoid-type MPM may be the progression of staging at
the first visit in the sarcomatoid-type.
PP01.53: PATTERN OF MALIGNANT PLEURAL
MESOTHELIOMA IN EGYPTIAN PATIENTS
Fatma M.A. Aou El-Kasem1, Abdel-Rahman M. Abdel-Rahman2,
Amr Demery2, Mohamed Rahoma3 , Rabab M. Gaafar1, Hala Aziz
Shokralla1, Maha Yehia1, Hisham Wahba4
Medical Oncology, National Cancer Institute, Cairo, EGYPT, 2Surgical Oncology, National Cancer Institute, Cairo, EGYPT, 3Surgical
Oncology, National Cancer Institute, Cairo, EGYPT, 4Radiology,
National Cancer Institute, Cairo, EGYPT
1
Objectives: A cancer registry was analyzed to determine the
clinico-pathologic characteristics affecting 194 malignant pleural mesothelioma (MPM) patients referred to National Cancer
Institute, Cairo University , Egypt from 2012 – 2015.
Methods: Retrospective review of patients with MPM presented to National Cancer Institute, Cairo University, Egypt;diagnosed between 2012 till 2015. Data regarding demographics,
histology, tumor staging and CT finding were obtained from all
patients. Pearson’s chi(2) and Fisher’s Exact tests were used for
statistical analysis.
Results: There were 194 cases of MPM referred to our Institute
during this period 2012 and 2015. We found that chest wall
invasion and pericardial infilteration ( p= 0.005), Transdiaphragmatic.extension (p= 0.016), presence of metastasis (p= 0.011)
and invasion of mediastinal structures (p=0.05) are significantly
correlated. Also, sex difference was statistically correlated with
pericardial infilteration ( p=0.026) and Transdiaphragmatic
extension (p= 0.021). Our patients’ age ranged from 15 to 76
years. Median age was 53 years. Ninity-five (49%)cases were
males. One hundred and nineteen (61.3%)cases were right sided. Pleural thickening was nodular in 131 (69.7%)cases, diffuse
in 46 (23.7%) cases and mass in 11 (5.7%) cases. Inter-lobar
fissure was thickened in 57 ( 29.4%) cases. Mediastinal Pleura
was affected in 72 (37.1%%) cases. Eighty-seven (44.8%) cases
had effusion. Ossification & calcification was detected in 8 (
4.1%) cases. Contraction of hemithorax was identified in 77
(39.7%) cases. Chest wall invasion was in 18 ( 9.3%) cases.
Pulmonary nodules were detected in 37 ( 19.1%). Metastases
were detected in 9 ( 4.6%) cases.
Conclusion: There is statistical significant correlation between
Chest wall invasion and pericardial infilteration ( p= 0.005),
Transdiaphragmatic.extension (p= 0.016), presence of metastasis (p= 0.011)and invasion of mediastinal structures (p=0.05).
Also, sex difference was statistically correlated with pericardial
infilteration ( p=0.026) and Transdiaphragmatic extension (p=
0.021). Early diagnosis and aggressive treatment for MPM
patients with chest wall invasion are recommended because of
high incidence for metstasis.
Keywords: positive rate, Pleural effusion, Malignant pleural
mesothelioma, histological type
iMig2016.ORG
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ABSTRACT BOOK
The number of deaths from mesothelioma in the years 20002012 among men was three times lower than in Italy, twice less
than in Germany and Sweden.
PP01.54: ASBESTOS CONSUMPTION AND
PLEURAL MESOTHELIOMA MORTALITY IN POLAND
IN COMPARISON WITH OTHER EUROPEAN
COUNTRIES
Conclusion: Conclusion The estimated peak of mesothelioma
cases appeared to be delayed in Poland in comparison with
that observed in the United Kingdom, likely due to the peculiar
Poland asbestos consumption curve. Acknowledgements: This
study was supported by the Ministry of Economy, Programme
for Asbestos Abatement in Poland 2009–2032.
Małgorzata Krowczynska1, Ewa Wilk1, Agnieszka Skubiszewska2, Anna Minkiewicz2, Gabriela Oledzka2
Department of Geoinformatics And Remote Sensing, University
of Warsaw, Warsaw, POLAND, 2Department of Medical Biology,
Medical University of Warsaw, Warsaw, POLAND
1
Keywords: asbestos consumption, malignant pleural mesothelioma,
Objectives: Malignant pleural mesothelioma (MM) is an
disease which is almost exclusively due to inhalation of
asbestos fibers. The protracted latent period of MM means
that its incidence has continued to rise across Europe after
the introduction of restrictions on asbestos use. Generally,
the mortality curve for asbestos-related cancers follows the
asbestos consumption curve with a lag of about 50 years. The
incidence of mesothelioma has increased off since 2012 and
steadily increasing in Poland. Like in most other industrial countries asbestos consumption increased in the twentieth century
in Poland. There are no asbestos deposits suitable for industrial exploitation in Poland therefore manufacture of asbestos
products in the years 1960 to 1997 was based on raw asbestos
imported mainly from the Russia and Africa. The decade 1970s
was the period of peak production of asbestos products, with
an annual consumption of about 100 thousand tons of raw
asbestos. We expecting at in Poland, the mortality peak of will
be reached only around 2020. The objectives of the present
study were: (i) compare of asbestos consumption in selected
European countries (ii) compare of the incidence of malignant
pleural mesothelioma (MM) in selected countries among men
and women in the years 2000 to 2012.
PP01.56: BLOOD DNA METHYLATION CHANGES IN
Elisabetta Casalone1, Simonetta Guarrera1, Marta Betti2,
Daniela Ferrante3 , Cornelia Di Gaetano1, Clara Viberti1,
Alessandra Biasi2, Sara Tunesi3 , Caterina Casadio4 , Francesco
Ardissone5, Enrico Ruffini6 , Roberta Libener7, Roberto
Guaschino8 , Ezio Piccolini9, Dario Mirabelli10, Corrado
Magnani11, Irma Dianzani12, Giuseppe Matullo1
Department of Medical Sciences, Human Genetics Foundation
and University of Turin, Torino, ITALY, 2Department of Health
Sciences, University of Piemonte Orientale, Novara, ITALY, 3Department Translational Medicine, CPO-Piemonte and Unit of
Medical Statistics and Epidemiology, Novara, ITALY, 4Thoracic
Surgery Unit, Azienda Ospedaliero-Universitaria‘‘Maggiore della
Carità’’,University of Piemonte Orientale, Novara, ITALY, 5Department of Clinical And Biological Sciences, Chest Surgery,
University of Turin, Orbassano, ITALY, 6Department of Oncology,
University of Turin, Torino, ITALY, 7Pathology Unit, SS.Antonio e
Biagio General Hospital, Alessandria, ITALY, 8Transfusion Centre,
Azienda Ospedaliera Nazionale SS, Antonio e Biagio e Cesare
Arrigo, Alessandria, ITALY, 9Santo Spirito, Hospital, Casale
Monferrato, ITALY, 10Unit of Cancer Epidemiology, CPO-Piemonte
and University of Turin and Interdepartmental Center for Studies
on Asbestos and other Toxic Particulates “G. Scansetti”, University of Turin, Torino, ITALY, 11Department Translational Medicine,
CPO-Piemonte and Unit of Medical Statistics and Epidemiology
and Interdepartmental Center for Studies on Asbestos and other
Toxic Particulates “G. Scansetti”, University of Turin, Novara,
Torino, ITALY, 12Department of Health Sciences, University of
Piemonte Orientale; Interdepartmental Center for Studies on
Asbestos and other Toxic Particulates “G. Scansetti”, University of
Turin, Novara, ITALY
1
Methods: This article is based on a selective review of the literature, along with data from the central database of the WHO
- Cancer Mortality Database.
Results: Objectives: The DNA-methylation status of various tissues
has been shown to be modulated by environmental exposures
and lifestyle. Epigenetic alterations have been also reported
in target MPM tissues suggesting an important role in the carcinogenic process. Evaluating whole blood DNA methylation as
a risk/diagnostic marker for cancer is of particular interest because whole blood DNA analysis is a noninvasive test and could
both reflects some common epigenetic changes in relation to
specific exposure and/or underlying specific immunological
response. We aimed to assess whether epigenome-wide DNA
methylation measured in white cells from whole blood samples
was associated with increased risk of MPM.
iMig2016.ORG
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ABSTRACT BOOK
Methods: We used the Illumina Human Methylation450 array
to measure methylation status in whole blood from a population-based case-control study sampled in four Northern Italian
towns: Casale Monferrato, Torino, Alessandria and Novara.
The analysis was performed on 183 cases and 194 controls for
which the level of asbestos exposure was assessed.
Results: The sample population was randomly split into two
groups: training and test set. In the training set three differentially methylated regions (DMRs) between cases and controls
were found (FDR adjusted p< 0.01), adjusting for gender, age,
asbestos exposure, and white blood cells percentage. We also
detected a global hypomethylation in cases respect to controls
(p = 0.007). We validated the DMRs prediction performance
in the test set using a Recursively Partionated Mixture Model
(RPMM) which clustered the samples in four classes on the
basis of the methylation profile of the three DMRs in the training
data. We assessed the increase in the performance of the methylation classifier by comparing receiver operating characteristic
(ROC) curves and the area under the curve (AUC) of two nested
models. The first included age, sex, centre and exposure as predictors; the second included also global and RPMM methylation
classes. The AUC was 0.75 (95% CI: 0.69-0.83) for the first
model and 0.78 (95% CI: 0.71-0.85) for the second, suggesting
a slight increase in the prediction performance given by the use
of DNA-methylation profiles.
Conclusion: Further statistical analyses are ongoing in order
to identify MPM methylation patterns taking into account
possible interaction between loci and asbestos exposure. We
suggest that epigenome-wide hypomethylation, and specific
genomic region methylation profiles of DNA detected in whole
blood MPM samples may be useful to further improve MPM risk
estimation, in addition to traditional assessment of asbestos
exposure.
Keywords: asbestos, methylation changes, exposure
IN AFRICA
tonnages. South Africa mined all three types of commercially
viable asbestos, viz. chrysotile, amosite and crocidolite, while
the other two countries mined only chrysotile. South Africa was
the global leader in the production of crocidolite asbestos, the
fibre most strongly linked with the development of mesothelioma - it was in this country that the link was first established and
reported in 1960. South Africa was also the only commercial
producer of amosite, the fibre closely implicated in the current
British mesothelioma epidemic. Asbestos has been used in
Africa for at least the past 5000 years (the ancient Egyptians
used it in mummification) but, before the 20th century, its use
was small scale. The 1920s onwards showed massive increases in its use, especially in North Africa, Nigeria and Southern
Africa, but virtually every country south of the equator used
asbestos in measurable tonnages. The largest consumers were
(in decreasing order) Nigeria, Algeria, Egypt, Morocco, Zambia,
Ghana and Tunisia, apart from the producer-countries. African consumption of asbestos exceeded 20,000 metric tonnes
annually between 1960 and 2000. Asbestos is currently banned
in Algeria, Egypt, Gabon, Mozambique and South Africa. Apart
from South Africa, there are few quality epidemiological studies
on mesothelioma in Africa. A common theme is under-ascertainment, especially among blacks. Mesothelioma rates have
been increasing, but may be levelling. A recent analysis of
mesothelioma deaths from the WHO mortality database (1994
– 2008) showed that, out of the 83 countries with data, Africans
died the youngest, and South Africa was in the top 10 for cumulative mesothelioma deaths. It has been reported that South
Africa has the highest rate of mesothelioma in those <50 years.
For environmental mesotheliomas, both Egypt and South Africa
have higher rates in women and children.
Conclusion: On the African continent, population-based mesothelioma rates are available for only South Africa. These rates,
despite being amongst the highest in the world, are likely to be
underestimates of the true burden of disease. Explanations for
this include under-reporting of cases, missed and misdiagnosis
of mesothelioma, competing causes of death, and reduced longevity related partly to the HIV/AIDS epidemic. The longevity of
South Africans is lower than that in other mesothelioma-reporting countries, so a smaller percentage of those exposed survive
to ages where mesothelioma might develop. Scientific efforts in
other African countries need to be encouraged.
Keywords: South Africa, Egypt, asbestos, epidemiology
Gill Nelson1, Jim Tewaternaude2
School of Public Health, University of Witwaterrand, Johannesburg, SOUTH AFRICA, 2School of Public Health, University of
Cape Town, Cape Town, SOUTH AFRICA
1
Objectives: Many African countries have used asbestos after
the 1920s and hence, millions of people have been exposed to
it. We set out to describe the epidemiology of malignant mesothelioma in Africa.
LONG TERM SURVIVORS: A POPULATION BASED
STUDY (LUME STUDY)
Methods: We searched the published and grey medical literature, and consulted experts.
Laura Botta1, Annalisa Trama1, Diego Signorelli2, Claudia
Proto2, Marina Chiara Garassino2, Roberto Foschi1, Riccardo
Capocaccia2, Sandra Mallone3 , Roberta De Angelis3 , Valerio
Gennaro4 , Lucia Benfatto4 , Cecilia Francesca Lando4 , Barbara Dallari5, Dario Consonni5, Carolina Mensi5, Enzo Merler6 ,
Vittoria Bressan6 , Manuela Gangemi7, Carol Brentisci7, Dario
Mirabelli7, Antonella Stura7, Antonio Romanelli8 , Cinzia Storchi8 ,
Elisabetta Chellini9, Francesca Battistini9, Adele Caldarella10,
Results: Although nine countries were listed as producers,
the only producers of significant amounts were South Africa,
Zimbabwe and Swaziland who, together, accounted for 99.8%
of asbestos production on the continent. The three countries
mined, respectively, 47%, 43% and 9% of the continent’s
iMig2016.ORG
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ABSTRACT BOOK
Cristiana Pascucci11, Mario Cocchioni11, Fabrizio Stracci12,
Maria Saba Petrucci12, Valeria Ascoli13 , Italo Angelillo14 , Daniela
Feola14 , Rosario Tumino15, Graziella Frasca16 , Maria Concetta
Giurdanella16 , Giovanna Tagliabue17, Paolo Contiero2, Gemma
Gola18 , Mariangela Corti18 , Francesca Bella19, Anna Clara Fanetti20, Salvatore Sciacca19, Antonio Ziino19, Rosanna Cusimano21,
Rosalba Amodio22, Roberto Piro23 , Pina Candela24 , Tiziana Scuderi24 , Claudia Cirilli25, Lucia Mangone25, Massimo Vicentini25,
Marcello Tiseo26 , Maria Michiara27, Anita Rimanti27, Anna Maria
De Giorgi26 , Paolo Sgargi27, Fabio Falcini28 , Orietta Giuliani28 ,
Rosa Vattiato28 , Francesco Forastiere29, Elisa Romeo29, Mario
Fusco30, Maria Francesca Vitale30, Silvano Piffer31, Roberto Vito
Rizzello31, Gemma Gatta1
Preventive And Predictive Medicine, Evaluative Epidemiology,
Fondazione IRCCS Istituto Nazionale dei Tumori, Milan, ITALY, 2Fondazione IRCCS Istituto Nazionale dei Tumori, Milan,
ITALY, 3Istituto Superiore di Sanità, Rome, ITALY, 4IRCCS, San
Martino, IST-Cancer Inst., Genoa, Italy; Registro Mesoteliomi,
Liguria, Genoa, ITALY, 5Registro Mesoteliomi della Lombardia,
Milan, ITALY, 6Registro Regionale Veneto dei casi di mesotelioma,
Padua, ITALY, 7Registro dei Mesoteliomi del Piemonte, Turin,
ITALY, 8Registro Mesoteliomi dell’Emilia Romagna, Reggio Emilia,
ITALY,9Centro Operativo Regionale Mesoteliomi della Toscana,
Florence, ITALY, 10Registro Tumori della regione Toscana, Florence, ITALY, 11Registro Mesoteliomi delle Marche, Camerino,
ITALY, 12Registro Mesoteliomi dell’Umbria, Perugia, ITALY, 13Department of Radiological, Oncological and Anatomo-Pathological
Sciences, Sapienza University, Rome, ITALY, 14Registro Mesoteliomi della Campania, Naples, ITALY, 15Centro Operativo Regionale
della Sicilia, Ragusa, ITALY, 16Registro Tumori, ASP, Ragusa,
ITALY, 17Fondazione IRCCS Istituto Nazionale dei Tumori, Registro
Tumori di Varese, Milan, ITALY, 18Cancer registry, Province of
Como, Como, ITALY, 19Integrated Cancer Registry of Catania-Messina-Siracusa-Enna, Catania, ITALY, 20 Cancer registry, Province of
Sondrio, Sondrio, ITALY, 21Cancer Registry of Palermo and province; Azienda Provinciale di Palermo, Palermo, ITALY, 22Cancer
Registry of Palermo and province, Palermo, ITALY, 23U.O. Pneumology ASMN IRCCS, Reggio Emilia, ITALY, 24Cancer Registry,
Province of Trapani, Trapani, ITALY, 25Epidemiology Unit, Azienda
Unità Sanitaria Locale, Reggio Emilia; Arcispedale Santa Maria
Nuova- IRCCS, Reggio Emilia, ITALY, 26Oncologia Medica, Azienda Ospedaliero- Universitaria di Parma, Parma, ITALY, 27Cancer
Registry of Parma, Parma, ITALY, 28Cancer Registry of Romagna,
Meldola, ITALY, 29Department of Epidemiology, Lazio Regional
Health Service, Rome, ITALY, 30Cancer Registry, Campania, c/o
ASL Napoli 3 Sud, Naples, ITALY, 31Cancer Registry of Trento,
Trento, ITALY
1
tumour with very poor prognosis, 50% of them dying within 9
months from diagnosis and survival time trends did not show
improvements over the last decades. However, the RARECARE
project observed long term survivors (LS=patients alive >3
years after diagnosis) suggesting the presence of milder phenotypes with a different prognosis and inspiring a dedicated
population-based observational study: long-term survivors in
pleural mesothelioma (LUME).
Methods: We collected from 26 Italian cancer registries all the
MPM newly diagnosed cases in the period 2003-2008 with cito-histological confirmation and with complete follow-up. In order to reproduce a correct representation of the Italian situation
we selected all the LS of the 26 registries and randomly sampled short survivors in each registry. The selected population
data included 2,475 MPM cases retrospectively collected. To
compare the differences detected in each variables distribution
between LS and short survival patients we use a χ2 test. The
Cox model assessed the prognostic value of selected variables.
Based on the results of each univariate model we selected the
variables included in the multivariate model.
Results: In our population the proportion of LS was 11%. The
χ2 test defined that the LS had an higher proportion of young,
female and epithelioid cases compare to short survivors. 55%
of the LS had localized stage vs 44% of the short survivors,
even if the proportion of missing stage is similar. Furthermore,
bimodal/multimodal treatment was more frequent among LS
(27% vs 11%). Interesting, 17% of LS (mean age 72) did not
received any treatment, suggesting the identification of an
indolent subgroup of cases. The Cox model showed age, sex,
histotype, cTNM, treatment as significant prognostic variables
(see the table below). Multivariate model
Variables
HR
p-value
Gender
Male
1
Female
0.86
0.001
0-54
0.74
<0.001
55-64
0.93
0.173
65-74
1
75+
1.29
<0.001
Mesothelioma NOS
1.09
<0.001
Sarcomatoid mesothelioma
1.35
<0.001
Epithelioid mesothelioma
1
Biphasic mesothelioma
1.78
Age class
MPM histotype
<0.001
Clinical TNM
T1/T2, N0/N1 and M0
1
T3, N0/N1 and M0 or T1/T2,
1.18
N2 and M0
0.007
T4 or M1 or N3
1.53
<0.001
Missing
1.15
0.038
Bimodal/multimodal
0.70
<0.001
Chemotherapy only
1
Surgery only
0.89
0.215
None/best supportive care
1.49
<0.001
Missing
1.12
0.275
Treatment
Diagnostic imaging
Only RX
1
Imaging (PET, TAC or RMN)
0.98
0.838
Conclusion: Our large population study confirms the results of
many clinical studies. Comparing with other population-based
studies we collected more detailed clinical variables on stage,
diagnostic procedures and treatment. Gender, age, histotype,
iMig2016.ORG
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ABSTRACT BOOK
type of treatment and stage were associated with prognosis.
Further statistical analysis are ongoing, including hospital of
diagnosis/treatment, immunohistochemistry and specific chemotherapy treatment. We also confirm the presence of MPM
long-term survivors (11%). The identification of their biological
features, which is one of the objectives of the LUME project,
may help clinicians in the definition of treatment.
in Southern Brazil are affected by inaccuracy and miscoding
regarding cancer site and histology. Moreover, medical records
are a poor source of information of industry and occupation.
This preliminary information support the need of a mesothelioma registry in Southern Brazil, and probably also in other parts
of the country.
Keywords: Brazil, cancer registry, mesothelioma incidence
Keywords: Population based study, Long term survivors, epidemiology, Prognostic factors
PP01.60: EPIDEMIOLOGY OF MESOTHELIOMA
IN SOUTHERN BRAZIL: A REALITY STILL TO BE
STUDIED
IN YOUNG PEOPLE
Ahmed El Bastawisy1, Maha Yahia1, M Rahouma2, Omnia
Aboelazm3 , Jaylan Ahmed4
National Cancer Institute, Cairo University, Cairo, EGYPT, 2Cardiothoracic Surgery, Weill Cornell Medicine, New York, AL, UNITED STATES OF AMERICA, 3Medical Biostatistics, National Cancer
Institute, Cairo University, Cairo, EGYPT, 4Clinical Pharmacy,
Baheya Cancer Center, Giza, EGYPT
1
Francisco José Koller , Leila Maria Mansano Sarquis , Luciana
Puchalski Kalinke1, Nen Nalú Alves Das Mercês1, Maria De
Fátima Mantovani1, Dario Consonni2, Carolina Mensi2
1
1
Escola De Enfermagem, Universidade Federal do Paraná, Curitiba, BRAZIL, 2Department of Preventive Medicine, Fondazione
IRCCS Ca’ Granda - Ospedale Maggiore Policlinico, Milan, ITALY
1
Objectives: Brazil is currently one of the largest producer of
chrysotile in the world. Mesothelioma occurrence in Brazil is
apparently low. However, identification of cases of mesothelioma in Brazil is difficult because of miscoding of diagnosis
and cause of death. For this reason, we are implementing a
mesothelioma registry in Curitiba (Paranà). Our aim is to show
preliminary data on mesothelioma occurrence in the Southern
Brazil using existing sources. Southern Brazil includes the
States of Paranà (PR, 10.4 million people), Santa Catarina (SC,
6.7 millions), and Rio Grande do Sul (RS, 11.2 millions).
Methods: We extracted data from the website of the National
Institute of Cancer in Brazil (INCA, https://irhc.inca.gov.br/
RHCNet/), containing data coming from hospital-based cancer
registries (RHC). We selected records of cancer of the pleura
(ICD-10: 38.4) or histology of mesothelioma regarding patients
residing in any of the three States in the period 2001-2013. We
describe clinical characteristics, demographics, and information on occupation.
Results: We identified 199 potential cases. We excluded
records unrelated to mesothelioma (26 adenocarcinomas,
22 lymphomas, 9 carcinomas, 7 sarcomas, and 6 with other
morphology). We also excluded 70 records with a generic definition of “malignant neoplasia”, because of a somewhat unusual
gender/age distribution (several women aged <50 years). Of the
57 remaining cases with histology of mesothelioma, 10 were
epithelioid, 4 fibrous, 1 biphasic, and 42 not otherwise specified. Notably, 14 of them were incorrectly coded as lung cancer
(ICD-10: C34). We found 17 cases (11 M, 6 F) from Paranà, 13
(6 M, 7 F) from SC, and 27 (22 M, 5 F) from RS. Regarding the
occupational activities performed in the last three months,
30 (53%), had no information. The rest of subjects had been
employed in various sector, including construction, metalmechanic, transport, and agriculture.
Conclusion: Existing hospital data regarding mesothelioma
Objectives: malignant pleural mesothelioma ( MPM) is characterized by long latency period between exposure to asbestos
and development of the disease so we hypothesize that MPM in
the young has different characteristics
Methods: This is a retrospective study including all eligible
patients with malignant pleural mesothelioma presenting to
National Cancer Institute, Cairo University during the period
from 2008 to 2013.
Patients were divided into two groups: Group 1: patients aged
≤ 45 years. Group 2: Patients aged > 45 years. Both groups
were assessed regarding different clinic-pathological features
.Primary Objectives: comparison of different epidemiological
features of both groups. Secondary Objectives: Assessment
of clinical response (CR), progression free survival (PFS) and
overall survival (OS) in both groups
Results: 102 Patients were included with median follow up of
14.4 months. Group (1) included 35 patients with mean age 40±
3.65 years (31 to 45 years).Group (2) included 67 patients with
mean age of 58.6± 8.5 years (46 to 87 years). 68% of group
(1) came from endemic areas which is significantly higher than
group (2): (35.8%), p = 0.02. History of Asbestos exposure was
highly significantly different between the 2 groups, 77.1% in
group (1) versus 38.8% in group (2), p < 0.001. Other factors
showed no significant differences between the two groups.
Overall clinical response (CR+PR) was 20% in group (1) versus
17.9 % in group (2). P=0.7. There was a trend towards longer
median PFS in young patients, (19.8 ± 8.4 versus 6.9 ± 1.4
months). p = 0.09. The median OS of young patients is significantly longer (20.6± 6.3 months) than older patients (11.4 ±
3.6).p = 0.05.
Conclusion: Mesothelioma in the young is more sensitive to
asbestos exposure, has better OS and likely a different disease
entity which needs further studies to understand its underlying
biological features.
Keywords: Malignant pleural mesothelioma, young
iMig2016.ORG
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ABSTRACT BOOK
IN AUSTRIA: DATA FROM THE AUSTRIAN
MESOTHELIOMA INTEREST GROUP DATABASE
Mir A. Hoda1, Thomas Klikovits1, Yawen Dong2, Paul Stockhammer2, Madeleine Arns3 , Peter Schenk3 , Wolfgang Pohl4 ,
Christian Geltner5, Michael Studnicka6 , Peter Cerkl7, Martin
Flicker8 , Josef Eckmayr9, Horst Olschweski10, Gertrude Blazek4 ,
Klaus Kirchbacher11, Peter Errhalt12, Bernhard Baumgartner13 ,
Helmut Popper10, Josef Bolitschek14 , Barbara Machan15, Walter
Klepetko1
Division of Thoracic Surgery, Medical University of Vienna,
Vienna, AUSTRIA, 2Thoracic Surgery, Medical University of Vienna, Vienna, AUSTRIA, 3LKH Hochegg, Hochegg, AUSTRIA, 4KH
Hietzing, Vienna, AUSTRIA, 5Pneumology, Klinikum Klagenfurt,
Klagenfurt, AUSTRIA, 6Pneumology, Paracelsus University Salzburg, Wels, AUSTRIA, 7Pneumology, LKH Hohenems, Hohenems,
AUSTRIA, 8LKH Leoben, Leoben, AUSTRIA, 9LKH Wels, Wels,
AUSTRIA, 10Medical University Graz, Graz, AUSTRIA, 11Hospital
Wilhelminen, Vienna, AUSTRIA, 12LKH Krems, Krems, AUSTRIA,13LKH Vöcklabruck, Vöcklabruck, AUSTRIA, 14LKH Elisabethinnen Linz, Linz, AUSTRIA, 15AUVA RZ Tobelbad, Tobelbad,
AUSTRIA
1
but aggressive tumor originating from the pleural cavity with
strong association to previous asbestos exposure. Despite
tremendous efforts in early diagnostics and therapeutic approaches, outcome for MPM patients remains dismal. In order
to determine demographics, diagnostics, therapeutic strategies
and prognosis of MPM patients in Austria, the Austrian Mesothelioma Interest Group (AMIG) was initiated in 2011. We intend
to report on the current data from the AMIG MPM database
Methods: A prospective cohort study starting from January
2011 was conducted. Follow up was completed until July 2015
Results: 203 patients with histologically confirmed MPM were
included. There were 162 male and 41 female patients with a
mean age of 67.1 (SD ± 11.4) years at the time of diagnosis.
Asbestos exposure was confirmed in 103 (50.1%) patients. 158
(77.8%) patients were diagnosed with pleural biopsy, 18 (8.9%)
with pleural puncture and 27 (13.3%) with other approaches. 190
(93.6%) patients had a chest computed tomography (CT), 120
(59.1%) received additional positron emission tomography (PET)/
CT and 20 (9.9%) a PET. Histological subtype revealed epithelioid
in 133 (65.5%), sarcomatoid in 15 (7.4%), biphasic in 28 (13.8%),
desmoplastic in 4 (2.0%) and not otherwise specified in 23
(11.3%) patients. 34 (16.7%) patients received best supportive
care only, 59 (29.1%) chemotherapy (CHT) alone, 3 (1.5%) radiotherapy (RT) alone, 19 (9.4%) CHT/RT, 1 (0.5%) surgery alone
and 72 (35.5%) curative surgery within multimodality treatment.
Median overall survival was 17.9 (95% CI, 14.6-21.3) months. 1-,
3- and 5-year overall survival was 66%, 26% and 19% and was
significantly better in patients undergoing surgery within multimodality treatment (5-year survival 14% vs 35%, p=0.001).
Conclusion: Diagnostic and therapeutic approaches according
to international guidelines are homogenous among different
centers in Austria. Patients undergoing multimodality treatment
including surgery had a favorable outcome.
PP01.63: HISTOPATHOLOGICAL REPORTING OF
MESOTHELIOMA RESECTION SPECIMENS
David A. Moore1, David Francis2, John Le Quesne1, Cathy
Richards1
Histopathology, University Hopsitals Leicester NHS Trust, WW,
UNITED KINGDOM, 2University of Leicester, RH, UNITED KINGDOM
1
Objectives: University Hospitals Leicester NHS Trust is a
leading centre for mesothelioma surgery and as such the histopathology department has developed considerable experience
in reporting resection specimens. The aim of this study was to
collate data from this valuable archive of cases to test the quality of diagnostic reporting for these specimens and to describe
their typical pathological profile in one of the few large cohorts
of these cases held in a single centre.
Methods: All mesothelioma resections from a single surgical
centre over a 5 year period were reviewed and the data was
collated. The immunoprofiles of the tumours were reviewed and
demographic data was also collected.
Results: 218 mesothelioma resections were received in the
pathology department over a 5 year period and were included in
the study. The male to female ratio was 181:37 and the median
age was 67 years. Of the 218 cases received there were 179
extended pleurectomy-decortications, 15 extrapleural pneumonectomies and 23 local resections. 78% of resected tumours
were of the epithelioid mesothelioma subtype. The majority
of cases included pericardium, diaphragm, lung parenchyma
and lymph nodes. On TNM staging over half of all cases were
tumour stage T3 and over half were nodal stage N2. Eight of the
samples (1 in 27) were sufficiently diagnostically challenging to
require external expert pathology review.
Conclusion: Pathology specimens from radical mesothelioma surgery are complex and challenging cases which require
specialist reporting. The data presented represents the first
description of the pathological characteristics from a series of
this scale from a single surgical centre.
Keywords: pathology, staging, resection
PP01.64: INHERITED PREDISPOSITION TO
Marta Betti1, Barbara Pasini2, Elisabetta Casalone3 , Alessandra Biasi1, Anna Aspesi1, Renzo Boldorini4 , Caterina Casadio5,
Enrico Colombo6 , Laura C. Gironi6 , Daniela Ferrante7, Federica
Grosso8 , Simonetta Guarrera3 , Antonella Maffè9, Dario Mirabelli10, Paola Ogliara2, Luisella Righi11, Simonetta Rosato12, Daniela
Turchetti13 , Sara Miccoli13 , Valeria Ascoli14 , Roberta Libener15,
Caterina Dianzani16 , Mauro Papotti11, Corrado Magnani17,
Giuseppe Matullo3 , Irma Dianzani18
Department of Health Sciences, University of Piemonte Orientale, Novara, ITALY, 2AOU Città della Salute e della Scienza di To1
Keywords: database, epidemiology
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ABSTRACT BOOK
rino, SC Medical genetics U and Department of Medical Sciences,
Turin, ITALY, 3Department of Medical Sciences, University of Turin;
Human Genetics Foundation,HuGeF, Turin, ITALY, 4Department
of Health Sciences, Section of Pathological Anatomy, University
of Piemonte Orientale, Novara, ITALY, 5Thoracic Surgery Unit,
Azienda Ospedaliero-Universitaria‘‘Maggiore della Carità’’,
Novara, ITALY, 6Dermatology Clinic, Department of Clinical
and Experimental Medicine, University of Piemonte Orientale,
Novara, ITALY, 7CPO-Piemonte and Unit of Medical Statistics and
Epidemiology, Department of Translational Medicine, University of Piemonte Orientale, Novara, ITALY, 8Division of Medical
Oncology, SS.Antonio e Biagio General Hospital, Alessandria,
ITALY, 9Molecular Genetics and Biology, Santa Croce e Carle,
Cuneo, ITALY, 10Unit of Cancer Epidemiology, CPO-Piemonte
and University of Turin; Interdepartmental Center for Studies on
Turin, Turin, ITALY, 11Department of Oncology, University of Turin
at San Luigi Hospital, Orbassano, Torino, ITALY, 12Department of
Obstetric, Gynecologic and Paediatric, Section of Clinical Genetics, Arcispedale S.Maria Nuova, Reggio Emilia, ITALY, 13Medical
Genetics, Policlinico Sant’Orsola-Malpighi, Bologna, ITALY, 14Department of Radiological, Oncological and Pathological Sciences,
Sapienza University, Rome, ITALY, 15Pathology Unit, SS.Antonio
e Biagio General Hospital, Alessandria, ITALY, 16Department
of Dermatology, “Campus Biomedico”, University of Rome,
Rome, ITALY,17CPO-Piemonte and Unit of Medical Statistics and
Epidemiology, Department of Translational Medicine, University
of Piemonte Orientale; Interdepartmental Center for Studies on
Turin, Novara, ITALY, 18Department of Health Sciences, University
of Piemonte Orientale; Interdepartmental Center for Studies on
Turin, Novara, ITALY
Objectives: BAP1 (BRCA1-Associated Protein 1) germline mutations predispose to a cancer-prone syndrome (MIM#614327)
that includes mesothelioma, cutaneous melanoma, uveal
melanoma and other cancers. This co-occurrence suggests that
these tumors share a common stepwise carcinogenic pathway.
Methods: To evaluate this hypothesis and to better characterize this syndrome, we collected 40 families classified into three
groups: 6 families with both mesothelioma and melanoma, 23
families with melanoma (without mesothelioma) and features of
inherited cancer predisposition and 11 families with mesothelioma (without melanoma) and features of inherited cancer predisposition. BAP1 gene was sequenced in all families and the same
families were also studied for the most common melanoma
predisposition genes (i.e. CDKN2A, CDK4 , TERT, MITF and POT1)
to investigate if these genes may also confer susceptibility to
mesothelioma.
susceptibility and these tumors share key steps that drive carcinogenesis. It also suggests that other genes may be involved
in inherited predisposition to malignant mesothelioma. Exome
analysis has been performed in two multiplex families that did
not show mutations in any of the tested genes.
Keywords: BAP1, CDKN2A, germline mutation
PP01.65: CHARACTERIZATION OF INTERTUMOR
HETEROGENEITY IN MALIGNANT MESOTHELIOMA
Noushin Nabavi, Raunak Shrestha, Yuzhuo Wang, Colin Collins
Urologic Sciences, Vancouver Prostate Centre, Vancouver, BC,
CANADA
Objectives: Malignant mesothelioma is a rare and aggressive
cancer. Here, we analyze the genomic and transcriptomic landscape of 87 pleural mesothelioma tumors to identify clinically
actionable driver genes and dysregulated signaling pathways.
Our longerterm objective is to compare and contrast the molecular profile of pleural mesothelioma with that of peritoneal
mesothelioma to advance the existing and relatively archaic
chemotherapeutic standard of care approaches.
Methods: We used publically available genomic and transcriptomic data from The Cancer Genome Atlas (TCGA) project
consisting of 87 pleural mesothelioma patients. We used our
bioinformatics methods to identify driver genes for mesothelioma and to integrate genomic (single nucleotide variant and copy
number alterations) and transcriptomic (RNAseq gene expression) data with clinically actionable relevance using established
genedrug interaction databases. One of our developed bioinformatics models, HIT’nDRIVE, relates the alterations at the genomic level to downstream changes in the transcriptome level,
where likely effects are propagated through gene interaction
networks. It then prioritize driver genes that dysregulates large
portion of the transcriptome. Next we utilized our bioinformatics method, OptDis, which uses the driver genes to search other
neighbouring genes to the driver in the interaction network (collectively called as subnetworks) such that the transcriptomic
profile of the subnetworks correlate with the sample phenotype.
These subnetworks help elucidate the molecular mechanisms
and pathways dysregulated in the mesothelioma patients.
Results: In two out of six families with both mesothelioma and
melanoma we identified a BAP1 germline nonsense mutation
(c.1153 C>T, p.R385X) and a common pathogenic germline missense mutation (c.301G>T, p.G101W) in CDKN2A. In both cases
BAP1 and CDKN2A proteins were not expressed in the tumor
tissues, respectively, supporting loss of heterozygosity (LOH).
Moreover, a patient with multiple cutaneous amelanotic melanomas carried a BAP1 germline missense mutation (c.1700A>C,
p.D567A) that was predicted to be pathogenic and a duplication
of 5bp (c.-594dupCCCGT) in the BAP1 promoter region. Microsatellite analysis supports LOH in the tumor tissue.
Results: We found a heterogenous inter intratumor molecular
landscape doemalignant mesothelioma. Our analysis identified
15 driver genes with varying alteration frequencies across 87
pleural mesothelioma patients. Somatic mutations in BAP1,
LATS2 and TP53 are among the previously identified driver
genes in pleural mesothelioma. The driver genes in 87 patients
along with their clinical characteristics have been summarized
in the figure below. The common pathways amenable to clinical
targeting fall in antigen processing and presentation, innate
immunity, and pathogen recognition as well as autophagy and
JAKSTAT signaling. We will further confirm these in sequencing
data obtained from peritoneal mesothelioma tumors.
Conclusion: Our study suggests that CDKN2A, in addition to
BAP1, could be involved in the melanoma and mesothelioma
Conclusion: Here, we present the inherent heterogeneity of
pleural mesothelioma at both pathologic and genetic levels.
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ABSTRACT BOOK
We identified commonly altered genes in all mesothelioma
subtypes from copynumber alterations to mutations to expression. We also present the canonical pathways we identified
from these datasets in each pathologic subtype (epithelioid,
sarcomatoid, biphasic, and diffuse malignant) based on our
computational analyses. The common pathways amenable to
clinical targeting fall in antigen processing and presentation,
innate immunity, and pathogen recognition as well as autophagy and STATJAK signaling. These findings infer the need for
a multimodal therapeutic intervention to account not only for
the heterogeneity of individual patient’s pathologic and genetic
profile but also the unique properties of solid tumors and their
microenvironment. Treatment effectiveness does not only depend on identifying the distinct molecular profile of malignant
pleural mesothelioma on an individual patient basis, but also
validation of the biomarkers in cell culture and animal models.
For these purposes, we further aim to obtain mesothelioma cell
lines and establish patientderived xenograft mouse models for
further biomarker identification and validation.
Keywords: pleural mesothelioma, peritoneal mesothelioma,
sequencing, patient-derived xenografts
PP01.66: PATHOLOGY ANALYSIS FOR
MESOTHELIOMA STUDY IN THE UNITED
KINGDOM: CURRENT PRACTICE AND HISTORICAL
DEVELOPMENT
97% were pleural mesothelioma, in line with estimates from
the LSHTM authors that only four per cent of their cases were
peritoneal. The most prominent subtype was epithelioid (64%
of study cases but only 49% of ineligible cases). Biphasic and
sarcomatoid subtypes constituted 10% vs. 16% and 7% vs.
19% of study and ineligible cases, respectively, reflecting earlier deaths in those ineligible. Of recorded immunohistochemical
stains for mesothelial cell origin, Calretinin (95%) and CK 5/6
or CK5 alone (84%) were by far the most common. Calretinin
and CK 5/6 or CK 5 alone were also most sensitive and positive
in 92% of cases having surgical pathology report. 90% of cases
had at least one immunohistochemical marker for possible lung
carcinoma applied, with BER-Ep4 and TTF-1 the most common
at 68% and CEA at 58%. TTF-1 and CEA were positive in one
per cent or less of cases. Keratin markers performed largely
as expected; MNF116 was used much more often than is the
case in Canada and was positive in 136 of 139 (98%). CK7 was
negative in 19% and CK20 positive in 4%. Microscopic description was recorded in 655 of the 748 cases (87.6%), but electron
microscopy was never reported. In 13 cases pathology report
specified a different diagnosis with no mention of mesothelioma. In another 18, more than one diagnosis was given with no
clear diagnosis favored by the pathologist.
Conclusion: Pathology practice showed only minor differences
from those in North America and Australia. Inclusion of nonmesothelioma cases could in theory result in lower odds ratios
for asbestos exposure. The differences between those “eligible”
and “ineligible” were related to shorter survivals.
Keywords: epidemiology, mesothelioma, immunohistochemistry, pathology
Bruce W. Case
Pathology, Epidemiology, School of Environment, McGill University, Montreal, QC, CANADA
Objectives: 799 mesothelioma cases were reviewed to provide
an overview of current pathology diagnostic practice in the
United Kingdom. Results were also aimed at identifying factors
which could affect epidemiological studies of the same cases.
Methods: Investigators at the London School of Hygiene and
Tropical Medicine (LSHTM) ascertained 1732 male and 670
female mesothelioma cases diagnosed between 2005 and 2013
in the process of ongoing study as of May of 2013. Cases were
deemed ineligible for further epidemiological study if they had
died or were too ill to allow direct questionnaire or interview.
Pathology reports were obtained as part of the file for 953
male cases (55%) and 357 female cases (53%). Of these, a
two-thirds sample was evaluated for the current study (860
cases). 61 (7%) of these had been included on the basis of
cytology reports, many of which gave equivocal diagnoses (e.g.,
mesothelioma “possible” or “unlikely”); these were excluded.
Of the remaining 799, with tissue diagnosis reported, 748 had
pathology reports sufficiently detailed for evaluation. Reports
were examined for basis of diagnosis, differences between
study cases and ineligible cases, pathology characteristics, and
immunohistochemical and other tests used.
Results: Cases were born 1925-1977, with 527 (66%) between
1940 and 1949. 566 were male (70.8%) and 233 female.
Males were significantly younger at diagnosis (63 years vs. 67
for women), and born later (1942 vs. 1947) (mean, P<.0001).
PP01.67: VARIATIONS IN COPY NUMBER IN THE
MALIGNANT PLEURAL MESOTHELIOMA GENOME
Marieke Hylebos1, Ken Op De Beeck1, Guy Van Camp2, Jan P.
Van Meerbeeck3
Center for Oncological Research, University of Antwerp, Edegem,
BELGIUM, 2Center of Medical Genetics, University of Antwerp,
Edegem, BELGIUM, 3Thoracic Oncology, Antwerp University Hospital, Edegem, BELGIUM
1
Objectives: Despite improvements in the outcome of malignant pleural mesothelioma (MPM) with the advent of chemotherapy, the median overall survival remains only 1 year. This
poor prognosis leaves ample room for the application of novel
treatment strategies. In several other solid tumors, the identification of actionable genomic alterations has resulted in a
paradigm shift in treatment towards specific targets. In an effort
to identify these actionable targets in MPM, studies already
reported on its genomic background. In this respect, karyotype
analyses and comparative genomic hybridization techniques
demonstrated a complex set of chromosomal copy number variations (CNVs) in most MPMs. These techniques however have a
limited resolution compared to highly sensitive next-generation
sequencing platforms, which allow genome-wide detections in a
high-throughput manner.
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ABSTRACT BOOK
Methods: The genomes of a set of 21 MPMs and matched
normal samples were analyzed using low-pass whole genome
sequencing (LP-WGS). Following DNA-extraction, sequencing
libraries were formed and paired-end sequencing (2 x 100 bp)
was performed on the ‘HiSeq 1500’ (Illumina). To enable the detection of structural variants, we aimed at a mean genome-wide
coverage of 1-2x. The presence of CNVs in the samples was
analyzed using in-house developed analysis pipelines.
Results: Preliminary analyses of the copy number profiles of
the paired samples showed striking differences. Whereas no
CNVs could be observed in the normal samples, copy number
losses and gains were visible in the tumor samples. Moreover,
it was readily detectable that copy number losses occur more
frequently compared to copy number gains. Overall, copy
number losses were most often observed in chromosomes 1p,
3p, 4q, 6q, 9p, 13q and 22q. Beside genes such as BAP1, CDKN2A and NF2, which are often found to be mutated in MPM,
these regions include several other possibly interesting targets.
Conclusion: Using LP-WGS, CNVs were detected in the
MPM-genome. Although the overall pattern of CNVs is heterogeneous and no single CNV was present in all of the samples,
the CNVs seem to cluster in certain regions. Further analysis of
these regions is ongoing and will be presented at the meeting.
Keywords: Copy number variations, Next-generation sequencing, Malignant pleural mesothelioma
PP01.68: HOMOZYGOUS 9P21 (P16/CDKN2A)
DELETION IN MESOTHELIOMA–5 YEAR
RETROSPECTIVE SPECIALIST DIAGNOSTIC
PLEURAL PATHOLOGY AUDIT
Lauren Harries1, Jayne Holme2, Matthew Evison2, Richard
Booton2, Philip Crosbie2, Rajesh Shah3 , Paul Taylor4 , Clare
Hodgson5, Nick Telford5, Paul Bishop1, Anshuman Chaturvedi1
Histopathology Department, University Hospital of South
Manchester, Manchester, UNITED KINGDOM, 2Manchester
Thoracic Oncology Centre, University Hospital of South Manchester, Manchester, UNITED KINGDOM, 3Department of Surgery,
University Hospital of South Manchester, Manchester, UNITED
KINGDOM, 4Oncology Department, University Hospital of South
Manchester, Manchester, UNITED KINGDOM, 5Oncology Cytogenetics, Christie Hospital, Manchester, UNITED KINGDOM
1
Objectives: Detection of homozygous deletion of the p16/CDKN2A gene on chromosome locus 9p21 using fluorescence in-situ hybridisation (FISH) in mesothelial cells has been reported to
improve diagnostic certainty of malignant mesothelioma (MM)
while assessing mesothelial proliferations in morphologically
difficult pathology cases. Histopathology department at University Hospital of South Manchester (UHSM) together with cytogenetics department at Christie Hospital over the previous 24
months (since early 2013) validated and then introduced p16/
CDKN2A FISH analysis, as a supporting technique, into routine
diagnostic practice. This five-year retrospective audit sought
to examine the volume and range of thoracic/pleural specimens reported to have mesothelial proliferations, focussing on
provision of history of asbestos exposure, utilisation of immunohistochemistry (IHC) (including EMA and Desmin) and p16/
CDKN2A FISH analysis and the degree of diagnostic certainty in
reporting of these specimens.
Methods: Electronic search of hospital pathology database
identified all thoracic/pleural specimens (fluid and biopsy)
coded as having a ‘mesothelial’related pathology, over a fiveyear period (2010 to 2015). Pathology reports were reviewed
systematically to ascertain provision of the history of asbestos
exposure, specimen type, IHC use, utilization and results of p16/
CDKN2A FISH and the diagnostic morphological end-point.
Results: 516 (82%) of the 628 specimens identified were
included in final analysis (117 pleural fluid, 381 pleural biopsy,
6 resection, 5 frozen sections, 2 other biopsy, 7 aspirate
cytology). Specimens excluded were those reported to have
non-mesothelial pathology. 38 coronial post-mortems were also
excluded. History of asbestos exposure was not provided with
326 (63%) specimens, while it was available and positive in
166 (32%) and negative in 24.IHC was performed in 455 (88%)
specimens (EMA and/or Desmin used in 81 (15%)). p16/CDKN2A
FISH analysis was attempted on 79 (15%) specimens. Insufficient material in two instances precluded FISH study. There
was test failure with one specimen. Homozygous 9p21 ( p16/CDKN2A) deletion was detected in 53 (67%) of all tests performed.
Remaining 25 specimens tested were negative. Diagnostic bottom-line in the 516 specimens included - confirmed diagnosis of
MM (n=287; 55%); highly suspicious for MM (n=64); suspicious
but not diagnostic of MM (n=51);suggestive of MM (n=12);atypical mesothelial proliferation (n=52); benign mesothelial cells
(n=4); non-diagnostic (n=1); atypia mesothelial versus epithelial
(n=33); MM versus carcinoma not established (n=4); mesothelial benign versus malignant (n=1). Of the specimens reported as
MM, the tumour sub-type was epithelioid (n=199); sarcomatoid
(n=41); biphasic (n=27); other (n=3). No subtype was recorded
in 111 specimens.
Conclusion: Definitive morphological diagnosis of mesothelioma is challenging. There is no diagnostic IHC profile for
mesotheliomas. Identifying tumour infiltration into adjoining
chest wall fat/ organ is the morphological feature widely-accepted to conclusively establish the diagnosis. However, in the
routine clinical scenario, not uncommonly, material available
is either limited superficial biopsy tissue or an initial cytology
sample. New diagnostic biomarkers are therefore needed. In
our experience, detection of homozygous deletion of p16/CDKN2A gene by FISH increased diagnostic certainty, including
for diagnostic cytology specimens. p16/CDKN2A FISH analysis
should be considered in challenging cases of atypical mesothelial proliferations where fat infiltration cannot be identified and
also in cytology cases where this is the only specimen available.
Keywords: Mesothelial proliferation, FISH, Cellular pathology,
Homozygous 9p21 ( p16/CDKN2A) deletion
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ABSTRACT BOOK
PP01.69: MUTATION STATUS AND EXPRESSION OF
THE MICRORNA-PROCESSING RIBONUCLEASE-III
DICER1 IN MALIGNANT PLEURAL MESOTHELIOMA
Alina Jørnild1, Morten Andersen1, Jesper Ravn2, Jens B.
Sørensen3 , Claus B. Andersen1, Morten Grauslund1, Eric Santoni-Rugiu1
Department of Pathology, Dept. of Pathology, Rigshospitalet,
Copenhagen University Hospital, Copenhagen, DENMARK, 2Department of Thoracic Surgery, Dept. of Thoracic Surgery,
Rigshospitalet, Copenhagen University Hospital, Copenhagen,
DENMARK, 3Department of Oncology, Dept. of Oncology, Rigshospitalet, Copenhagen University Hospital, Copenhagen, DENMARK
1
Objectives: We and others recently reported that deregulated
expression of specific microRNAs (miRNAs) in malignant pleural
mesothelioma (MPM) may aid in the difficult differential diagnosis between MPM and benign reactive mesothelial proliferations (RMP), has prognostic significance, and depends at least
in part on epigenetic mechanisms [1-3]. The ribonuclease-III
Dicer1 plays a fundamental role in the biogenesis of miRNAs. In
other cancer-types, somatic hot-spot mutations affecting the DICER1-gene or up-/down-regulation of Dicer1-mRNA/protein
resulting in abnormal Dicer1-function and aberrant miRNA-biosynthesis with prognostic impact, have been described. Thus,
we investigated whether DICER1-mutations and/or deregulated
-expression may represent alternative mechanisms underlying
MPM’s deregulated miRNA-profiling.
Methods: Formalin-fixed paraffin-embedded tissue specimens
from a cohort of 78 surgically resected MPMs (pleurectomy/
decortication) receiving neoadjuvant cisplatin-pemetrexed
(40 epithelioid/38 biphasic; stage II-IV; period 2011-2015), 23
available patient-matched non-neoplastic pleuras (NNP), 10
adjacent non-invasive atypical mesothelial proliferations (AMP),
and 5 unrelated chemotherapy-naïve diagnostic MPM-biopsies
(DB; epithelioid), as well as 12 independent pneumothorax-induced RMP, were investigated. Possible hot-spot mutations
of DICER1-gene were analyzed on genomic DNA by nested PCR
and dideoxynucleotide-sequencing of exon 24-25 encoding
the catalytic RNase-IIIb domain. Dicer1-mRNA expression was
measured on total RNA by RT-qPCR normalizing to the internal
reference MRPL19 -gene. Dicer1-protein expression was assessed
by IHC using light microscopy and a modified semi-quantitative
H-score. Significant (P<0.05) differences in Dicer1-mRNA/-protein expression between independent or paired groups of
samples were detected by non-parametric Mann-Whitney and
Wilcoxon signed-rank tests, respectively, while non-parametric
Kruskal-Wallis ANOVA was used for comparing expression in
samples from different groups.
Results: Two patient-matched NNP-MPM pairs displayed
single-nucleotide-polymorphisms, but no somatic mutations
were identified in exon 24-25 of DICER1-gene in any of the MPM, NNP-, AMP-, DB- or RMP-samples analyzed, suggesting that
as opposed to other cancer-types these hot-spot mutations are
uncommon/absent in MPM. No significant difference (P >0.05)
of Dicer1-mRNA expression in MPM vs. NNP, AMP or RMP was
found. In contrast, significant upregulation of Dicer1-protein
(P<0.05) was detected in MPM as compared to NNP or RMP but
not AMP or DB. Comparable Dicer1 expression in MPM- and
DB-groups indicated no significant influence of chemotherapy.
A numerical (non-significant) trend of up-regulated Dicer1-mR-
NA and -protein was observed in epithelioid vs. biphasic MPM,
possibly reflecting the association between lower Dicer1 expression and poorer prognosis reported in some other cancers.
Moreover, numerically higher Dicer1 expression was observed
in MPM stage IV vs. earlier stages.
Conclusion: DICER1 catalytic domain’s hot-spot mutations
and Dicer1-mRNA expression level do not appear to be causal
mechanisms for aberrant miRNA-expression in MPM. However,
Dicer1-protein overexpression suggests a role for this ribonuclease in the miRNA-deregulation and possibly pathogenesis
of this cancer. Together with observations in other malignancies, these results imply that causes and effetcs of aberrant
Dicer1-function may depend on cancer type. Further research
on Dicer1 and other miRNA-processing pathway’s components
is required to clarify the mechanisms of miRNA-deregulation
in MPM. 1. Andersen M. et al., J Mol Diagn 16:418–30, 2014 2.
Andersen M et al., Anticancer Res 35:6223–9, 2015 3. Santoni-Rugiu E, Andersen M, Grauslund M, Current Biomarker
Findings 6:1-21, 2016
Keywords: mesothelioma, microRNA-regulation, Dicer1
PP01.70: A COMPARISON OF THE GENETIC
CHARACTERISTICS OF MURINE AND HUMAN
Sophie Sneddon1, Ian M. Dick1, Nicola Waddell2, John Pearson2, Richard J. Allcock3 , Bruce Robinson1, Jenette Creaney1
Perth, WA, AUSTRALIA, 2QIMR Berghofer Medical Research Institute, Brisbane, QLD, AUSTRALIA, 3University of Western Australia, School of Pathology and Laboratory Medicine, University of
Western Australia, Perth, WA, AUSTRALIA
1
Objectives: Next generation sequencing approaches have the
potential to result in new treatment strategies for cancer. A murine model of mesothelioma can be an ideal tool for analysing
the genetic changes of mesothelioma in finer detail and to allow
testing of targetable genetic lesions, provided it is similar to the
human disease. With the aim of ultimately taking advantage
of the established pipeline of testing new therapies in preclinical murine models of disease we have compared the exome
sequence of mesothelioma in patients and in asbestos-induced
tumours in mice.
Methods: Whole exome sequencing (WES) was performed
on primary mesothelioma cultures from 29 patient and 16
mouse effusions using the Ion Torrent Proton sequencer along
with normal tissue as controls. Somatic single nucleotide
variations (SNVs) were identified using VarScan2 and SomaticSniper software, pooled and annotated. Small regions of copy
number variation (CNV) were identified using ExomeCNV and
R. GISTIC2.0 and MutSigCV were used to identify significant
SNVs and CNV across all samples. Mutational signatures were
derived using SomaticSignatures in R.
Results: There was an average of 6.6 SNVs per Mb across
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ABSTRACT BOOK
the human and 9.7 per Mb in the murine samples with a high
number of C>T and G>A transitions. There was evidence for selection pressure of missense mutations, indicating the presence
of possible driver mutations. Both species showed a higher
proportion of copy number losses than gains. Evidence of a
possible mutational signature was identified. The most frequent
genetic change in both human and murine mesothelioma was
p16 loss. Genetic alterations in BAP1 and NF2 occurred at lower
frequency in mouse relative to human mesothelioma.
Conclusion: MM is a complex tumour with a number of genetic characteristics. Not only is there similarity in phenotypic
characteristics between asbestos-induced tumours in humans
and mice, there are similarities at the genetic level. This study
supports the use of the immune-competent murine model for
future mesothelioma genetics and treatment studies.
Keywords: bioinformatics, mutations, exome sequencing,
murine model
PP01.71: MUTATIONAL BURDEN AND CDKN2A
STATUS AS A ROBUST OUTCOME PREDICTOR FOR
RADICAL SURGERY IN MESOTHELIOMA
Annabel J. Sharkey1, Robert Hastings2, David A. Moore3 , John
Le Quesne3 , Morag Taylor4 , Phillip Quirke4 , Apostolos Nakas5,
David Waller5, Sara Busacca1, Dean Fennell1
Cancer Studies, University of Leicester, Leicester, UNITED KINGDOM, 2University of Leicester, Leicester, UNITED KINGDOM, 3Histopathology, University Hopsitals Leicester NHS Trust, Leicester,
UNITED KINGDOM, 4Leeds Institute of Pathology and Tumour
Biology, Leeds, UNITED KINGDOM, 5Glenfield Hospital, University
Hospitals of Leicester NHS Trust, Leicester, UNITED KINGDOM
1
Objectives: Mesothelioma is a heterogeneous cancer with
respect to its natural history, with a subset of patients exhibiting
relatively indolent cancer associated with long time to progression (TTP) and overall survival (OS) following surgical resection.
The aim of this study was to interrogate the extent of genomic
instability of this indolent phenotype through copy number
analysis in order to identify predictive molecular features.
Methods: 50 matched patients were identified from the Leicester Mesothelioma radical surgery tissue bank from 1999-2014.
OS of these 436 patients was 15 months (range 0-167 months).
Patients were categorized according to either long OS (>15
months) or short OS (<15 months). OS for long survivors (LS)
was 45.8 months (17.3-66.2 months), and short survivors (SS),
6.2 months (1.0-14.5 months). Patients were matched based on
all known clinic-pathological prognostic factors, and all were
epithelioid histological subtype. No patients received neoadjuvant chemotherapy and four patients, 2 in each group, were
treated with platinum/pemetrexed at progression. Somatic copy
number alteration (SCNA) was determined from FFPE extracted DNA using an array based platform (Affymetrix Oncoscan
v3). Putative SCNAs were identified using a circular binary
segmentation algorithm, and significant recurrent SNCAs were
identified by GISTIC algorithm. Total SCNA, or SCNA(N), was
estimated as a surrogate for mutational burden using NEXUS
Express v3.1. Test and validation cohorts comparing LS versus
SS were segregated and the area-under-the-curve (AUC) of
corresponding receiver-operator curves, defined.
Results: Twenty-eight matched patients were subgrouped
into a test cohort comprising 14 long survivors (LS) versus 14
short (SS). A validation cohort comprised 16 patients. SCNAs:
LS losses 6q25.3 (79%), 3p22.1 (64%), 1p12 (50%), 9p21.3
(43%), 22q11.23 (36%), 17q25.3 (29%) 16p11.2 (24%), 8p11.22
(21%), 6p25.3 (7%), 1q21.2 (7%). LS gains 22q11.23 (57%) and
16p11.1 (36%). SS losses 9p21.3 (CDKN2A) (79%), 22q11.23
(50%) 1q21.2, (36%),8p11.22 (14%). SS gains 22q11.23 (64%),
8p11.22 (29%) and 16p11.2 (29%). Median SCNA(N) was 96
(range 8-398) and negatively correlated with both OS and TTP
for all patients (OS; ρ -0.526, p=0.004, TTP; ρ -0.631 p=0.002).
Median SCNA(N) for LS was 74.5 (8-159) and SS was 113.5
(58-398). The cut off of SCNA(N) ≥90 most accurately predicted
long versus short OS and TTP (AUC(OS) =0.750 and AUC(TTP)
= 0.818). 9p21.3 (CDKN2A locus) loss predicted OS and TTP;
AUC(OS) = 0.719, AUC(TTP) = 0.657. Using either SCNA(N) ≥90
or the presence of CDKN2A loss (either homozygous or heterozygous), or both, improved the outcome prediction accuracy;
AUC (OS) = 0.806 and AUC(TTP) = 0.798. Having SCNA(N) ≥90
plus CDKN2A loss gave an AUC(OS) = 0.833 and AUC(TTP) =
0.838. In the validation cohort (median OS 56.3 months, range
3.5-152.3 months, median TTP 17.9 months, range 2.1- 145.4
months), CDKN2A loss did not correlate with mutational burden
or clinico-pathological features. However longer OS was associated with SCNA(N)≥90 plus CDKN2A loss (56.3 vs. 28.5 months
p=0.518), as was TTP (18.4 vs. 6.2 months p=0.058).
Conclusion: CDKN2A wild type mesothelioma tumours
harbouring <90 SCNAs may exhibit long TTP and OS following
radical surgery. Further validation of this potential prognostic
tool is required.
PP01.72: WHOLE-GENOME DNA METHYLATION
AND TRANSCRIPTOME CHANGES IN ASBESTOS
EXPOSED MET5A CELLS
Elisabetta Casalone1, Alessandra Allione1, Simonetta Guarrera1, Clara Viberti1, Barbara Pardini1, Marta Betti2, Cornelia
Di Gaetano1, Corrado Magnani3 , Irma Dianzani4 , Elisabetta
Aldieri5, Giuseppe Matullo1
Department of Medical Sciences, Human Genetics Foundation
and University of Turin, Torino, ITALY, 2Department of Health Sciences, University of Piemonte Orientale, Novara, ITALY, 3Department Translational Medicine, CPO-Piemonte and Unit of Medical
Statistics and Epidemiology and Interdepartmental Center for
Studies on Asbestos and other Toxic Particulates “G. Scansetti”,
University of Turin, Novara, Torino, ITALY, 4Department of Health
Sciences, University of Piemonte Orientale; Interdepartmental
Center for Studies on Asbestos and other Toxic Particulates “G.
Scansetti”, University of Turin, Novara, ITALY, 5Department of
Oncology, Interdepartmental Center for Studies on Asbestos and
other Toxic Particulates “G. Scansetti”, University of Turin, Torino,
ITALY
1
Objectives: Occupational and environmental asbestos expoiMig2016.ORG
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sure is the main determinant of MPM development, which is
characterized by a long latency period. Apart asbestos induced
mesothelial inflammatory processes that are thought to be
the basic mechanism underlying MPM development, there
are several other possible mechanisms through which asbestos may induce MPM, that have not been already explored.
Asbestos exposure may act to induce epigenetic errors leading
to the deregulation of gene expression. We investigated whether
a dose-response relationship exists between alterations of
DNA methylation profiles and asbestos exposure in a well-controlled in-vitro experimental setting, to determine the impact of
methylation changes on gene expression.
Methods: We measured whole genome expression and DNA
methylation levels on Met5A human mesothelial cell lines treated with increasing concentration of crocidolite and chrysotile
asbestos for 72 hours (0.5 ÷ 5.0 µg/cm2). Spearman’s rank
correlation was employed in order to examine the association
between DNA methylation or gene expression levels and asbestos fibre doses.
Results: The DNA methylation dose-response relationship
in crocidolite and chrysotile treated cells showed changes in
methylation levels of numerous CpG sites (rho = ±1, p<0.05).
Among the genes overlapping the set of CpGs associated with
asbestos dose, we identified several genes already described
with altered methylation patterns in MPM such as tissuewingless-type MMTV integration site family member 10B (WNT1
0B),NOTCH4 , homeobox A5 (HOX5), MutS protein homolog
2
(MSH2) and others. Expression analyses on mRNA levels
revealed a significant association with crocidolite and chrysotile dose for 950 and 575 transcripts, respectively (rho = ±1,
p<0.05). We recognized a large number of previously known as
well as new potential asbestos-related differentially expressed
genes and biological processes, among which we identified
genes that play a role in the regulation of cell fate, cell cycle,
cell growth and DNA damage repair. Noteworthy, there was the
statistically significant linear correlation between DNA methylation and mRNA expression observed for 14 genes in crocidolite
treated cells, including CDK6 , FANCB, and five genes in chrysotile exposed cells, such as IGFBP5 and ITGB5 (rho = ±1, p<0.05).
Conclusion: This study identified several interesting targets
for further investigation in relation to asbestos exposure and
represents an important step to understand if asbestos-induced
carcinogenesis may be related to DNA methylation changes.
Additional analyses are ongoing in order to replicate the signals
that have already been identified in MPM tissue and with a
relevant biological function and to evaluate the impact of these
processes in MPM development.
Keywords: cells treatment, espression changes, asbestos,
methylation changes
PP01.73: GENOMIC INTERROGATION OF A CLONAL
RECURRENCE OF PLEURAL MESOTHELIOMA 12.5
YEARS AFTER RADICAL SURGERY
Annabel J. Sharkey1, Robert Hastings2, David A. Moore3 , John
Le Quesne3 , Morag Taylor4 , Phillip Quirke4 , David Waller5, Apostolos Nakas5, Sara Busacca1, Dean Fennell1
Cancer Studies, University of Leicester, Leicester, UNITED
KINGDOM, 2University of Leicester, Leicester, UNITED KINGDOM, 3Histopathology, University Hopsitals Leicester NHS Trust,
Leicester, UNITED KINGDOM, 4Leeds Institute of Pathology and
Tumour Biology, Leeds, UNITED KINGDOM, 5University Hospitals
of Leicester, Leicester, UNITED KINGDOM
1
Objectives: Malignant pleural mesothelioma (MPM) remains
an incurable cancer. Little is known about the genotype-phenotype relationship that determines the natural history of
mesothelioma, particularly in the context of extremely indolent
mesotheliomas associated with exceptionally long times to
progression (TTP) following radical surgery. Molecular classification of such patients could facilitate rational stratification for
surgery to optimize outcomes. Accordingly we have conducted
paired genome-wide somatic copy number alteration analysis
of a patient who exhibited extreme TTP post extra-pleural pneumonectomy (EPP).
Methods: A 69 year old man underwent right EPP after 3
cycles of neoadjuvant chemotherapy for epithelioid MPM in
2002 stage pT3N0. No further oncological treatment was given
post-operatively. In 2015, 12.5 years after his original resection,
he developed chest wall recurrence and nodules in the remaining lung. A paired biopsy was taken to confirm a diagnosis
of recurrent MPM, and to allow comparable genetic analysis
pre-surgery and at disease progression. The array based
platform Affymetrix Oncoscan v3 was used to determine genome-wide somatic copy number alteration (SCNA) using FFPE
extracted DNA from both the initial resection specimen, and the
recurrence biopsy. SCNA number was determined using NEXUS
Express v3.1. In previous work (abstract #374) we identified
SCNAs specific to either long or short survival groups following
surgery for MPM using the GISTIC algorithm for segregation of
significant SCNAs.
Results: Total SNCA was 18 in the original tumour and 55 in
the recurrence sample, with 39 new SCNAs present in the recurrent tumour. Sixteen (88.9%) SCNAs were common between
the two tumours and are therefore likely to be clonal. Of those
genes commonly lost in our previous cohort of long survivors, 9
were found to be lost in this patient’s original tumour; ADAM3A
(homozygous loss), ZDHHC14 (heterozygous loss), TMEM242
(heterozygous loss), MIR3692 (heterozygous loss), CCK
(heterozygous loss), LYZL4 (heterozygous loss), and TP53 TG3,
TP53 TG3B and TP53 TG3C (loss of heterozygosity). Of the cosmic mutations commonly found to be altered in MPM (CDKN2A,
BAP1 and NF2) neither tumour harboured loss of CDKN2A, and
both harboured BAP1 heterozygous loss. A new heterozygous
loss of NF2 was found in the recurrence sample.
Conclusion: In summary, clonal progression of mesothelioma
is still possible after a decade of disease control. The mechanisms underlying dormancy are unknown. Bap1 loss does not
preclude exceptionally long time to progression in the context
of low mutational burden, and progression may have resulted
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from emergence of the NF2 driven subclone, coincident with increased mutational burden as suggested from parallel genomic
studies to be presented.
PP01.74: CDKN2A DELETION: A CLONAL
MUTATION IN MALIGNANT PLEURAL
MESOTHELIOMA DEVELOPMENT?
Luke Martinson1, Annabel J. Sharkey1, Barbara Ottolini1, David
A. Moore2, John Le Quesne2, Apostolos Nakas3 , David Waller3 ,
Jacqui Shaw1, Dean Fennell1
Cancer Studies, University of Leicester, Leicester, UNITED KINGDOM, 2Histopathology, University Hopsitals Leicester NHS Trust,
Leicester, UNITED KINGDOM, 3Glenfield Hospital, University
Hospitals of Leicester NHS Trust, Leicester, UNITED KINGDOM
1
Objectives: Intratumour heterogeneity is likely to contribute
to treatment failure in malignant pleural mesothelioma (MPM),
with the targeting of subclonal rather than clonal (ubiquitous)
drivers. The most common genetic alteration observed via a
single tumour biopsy, is homozygous deletion ofCDKN2A (p16/
p14ARF), observed in approximately 70% of cases. Determination of CDKN2A deletion as a clonal driver event is vital for both
future therapeutic intervention and predicting patient outcome
following treatment. Ongoing work in our group has indicated
that patients harbouringCDKN2A deletion may exhibit a poorer
clinical outcome following radical surgery (see Abstract 374).
Therefore, providing CDKN2A deletion is confirmed to be an early clonal event, the analysis of a single site pre-operative tissue
biopsy (or potentially a liquid biopsy) could be of prognostic
benefit to patients undergoing radical surgery. Here, we investigate the clonal origin of CDKN2A deletion in MPM using Droplet
Digital PCR (ddPCR) on multiregional tumour tissue.
Methods: Multiregional tumour tissue (5 spatially separated
anatomical regions) was obtained from patients undergoing
extended pleurectomy/decortication (EPD). Each patient additionally provided a blood sample to allow for the isolation of
lymphocyte DNA, to be used as a healthy genomic control. All
tumour regions were assessed for approximate tumour content
by a specialist thoracic histopathologist. ddPCR (Bio-Rad)
was conducted using 5ng template DNA. An in-house RPPH1
assay was multiplexed with the CDKN2A assay, to act as a copy
number reference (≈1 copy/haploid genome). H460 cell line
was used as a positive control for CDKN2A deletion (HD). 7 of
these samples were cross-validated for CDKN2A status using
the OncoScan® FFPE Assay platform (Affymetrix), using one of
the five tumour regions.
Results: 53% of patients (9/17) exhibited evidence of CDKN2A deletion (HD/Loss of heterozygosity (LOH)), which is
relatively consistent with current literature. Due to unavoidable
contamination from stromal (healthy) cells within each tumour
region, the threshold for CDKN2A deletion was set at 0.8/1 copy
of reference, with values below 0.8 indicating evidence of deletion. Oncoscan® analysis showed that the ddPCR results were
reproducible in all 7 cases. Samples with detected HD of CDKN2A via Oncoscan®, possessed an average copy number of 0.38
by ddPCR, compared to 0.78 for those with LOH
Conclusion: In all patients exhibiting evidence of CDKN2A deletion, this was observed across all 5 tumour regions, implicating CDKN2A deletion as a clonal event in MPM development.
These findings indicate that CDKN2A profiling from a single
tumour biopsy (or potentially a liquid biopsy) using ddPCR could
be viable, cost-effective and efficient in the clinical setting.
Additionally this data consolidates CDKN2A as not only an ideal
therapeutic target in future MPM treatment, but also allows CDKN2A status to be used as a possible prognostic marker for
patient outcome following surgery.
PP01.75: UPDATE: RECENT STUDIES EXAMINING
IMPACT OF BAP1 MUTATION ON MESOTHELIOMA
RISK AND IMPLICATIONS FOR MESOTHELIOMA
LITIGATION
Steven Kazan
Kazan McClain Satterley & Greenwood, A Professional Law Corporation, Oakland, UNITED STATES OF AMERICA
Objectives: Background: At iMig 2014 we discussed the
ethical issues surrounding BAP1 testing and its role in asbestos
lawsuits. 1. The case referenced in our poster (#P1.061) started
jury trial on January 5, 2016 and resolved on February 3, 2016,
just before closing arguments. The author of the affidavit cited
in our 2014 poster was withdrawn as an expert by the asbestos
cement pipe manufacturer which had retained him. Testimony
by one of his co-authors contradicted his claims of causality,
and the published medical literature also increasingly supports
the role of the gene/environment interaction in the oncogenesis
of mesothelioma in BAP1 cancer syndrome. An initial article had
hypothesized that BAP1 mutations alone might be sufficient to
cause mesothelioma. 2. However, since 2014, the majority of
the literature concludes that BAP1 mutations leave an individual
increasingly vulnerable to carcinogens like asbestos.
Methods: A PubMed literature review was conducted on all
peer reviewed English language articles indexed between 2014
and 2016, with keyword searching for germline BAP1 and mesothelioma. Twenty-seven articles were found. We then categorized the conclusions and charted the trends over time.
Results: Four articles were eliminated as irrelevant. Eleven
articles discussed exposure to environmental factors as a
cause of oncogenesis. Two articles discussed low exposure to
environmental factors as a cause of oncogenesis. One article
hypothesized that asbestos exposure might not be necessary
for the development of mesothelioma.3
Conclusion: Current literature strongly supports the conclusion that individuals with a germline BAP1 mutation are more
vulnerable to oncogenesis of certain tumors after exposure to
carcinogens. In the case of mesothelioma, this carcinogen is
asbestos. The utility of using an existing BAP1 mutation as a
defense to liability for mesothelioma is increasingly questionable. 1. Kazan and Kin. Hippocrates and BAP1 Genetic Testing
in Mesothelioma Litigation. Abstract P1.061, iMig 2014, p. 30.
2. Testa, et al. Germline BAP1 mutations predispose to malignant mesothelioma. Nature Genetics 43.10 (2011): 1022-1025.
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3. Carbone, et al. Combined Genetic and Genealogic Studies
Uncover a Large BAP1 Cancer Syndrome Kindred Tracing Back
Nine Generations to a Common Ancestor from the 1700s. PLoS
Genet 11.12 (2015): e1005633.
Keywords: litigation, BAP1, mutation
PP01.76: BAP1 VALUE IN DIAGNOSIS
Michele Carbone
STATES OF AMERICA
Objectives: BAP1 germline mutations cause malignant mesothelioma and other cancers, and BAP1 somatic mutations
are the most common genetic alteration in sporadic mesotheliomas. We investigated the potetnial value of BAP1 testing in
differentiating mesothelioma from other malignancies.
Methods: We used an array of different molecular techniques
and classical immunohistochemistry.
Results: By testing a large number of mesotheliomas and
human specimens we found significant differences that indicate
that ascertain the BAP1 status is useful in the differential diagnosis of mesothelioma.
are not well documented. This means that individual research
groups often repeat similar experiments, resulting in an excess
use of animals. In addition, unlike the human situation, it results
in a highly variable use of chemotherapy dosages across the
literature, with different biological effects. We set out to define
the maximum tolerated dose (MTD) for 10 classes of chemotherapeutics in mice using a comprehensive and systematic
approach and we investigated the effect of supportive care on
chemotherapy-induced toxicity.
Methods: Chemotherapy was administered to BALB/c mice
with doses escalated until the endpoints of weight loss >15%
or a clinical score >2 were met. A parallel series of experiments
tested MTDs in C57BL/6J mice and multiple dosing at MTD.
To determine the effect of supportive care, dexamethasone,
ondansetron and supplementary feed were used both as single
agents and in combination.
Results: For nine of the ten drugs tested, weight loss was the
dose-limiting toxicity, while clinical score determined the MTD
of gemcitabine. For some chemotherapeutics the MTD was
substantially higher than we commonly found in the literature.
There was a slight variability in MTD between mouse strains.
The tolerability of repeated cycles was drug-dependant. The use
of either nutritional supplement or ondansetron did not reduce
toxic effects of cisplatin chemotherapy while experiments with
dexamethasone are ongoing; the results will be reported at the
meeting.
Conclusion: BAP1 testing should always be conducted in mesothelioma patients for diagnostic and prognostic reasons
Conclusion: These data are a resource for future studies using
chemotherapy in mice and should reduce the number of mice
required for dose optimization experiments. Our studies also
suggest that supportive care can reduce toxic side effects and
increase the MTD.
Keywords: Malignant mesothelioma, BAP1, pathology, differential diagnosis
Keywords: Supportive care, chemotherapy, maximum tolerated dose
PP01.77: A SYSTEMATIC INVESTIGATION OF THE
MAXIMUM TOLERATED DOSE OF CYTOTOXIC
CHEMOTHERAPY WITH SUPPORTIVE CARE IN
MICE
PP01.78: CANCER CHEMO IMMUNOTHERAPY EXPLOITING THE IMMUNOGENIC POTENTIAL OF
CYCLOPHOSPHAMIDE
Wayne J. Aston1, Danika E. Hope1, Scott Fisher1, Anna Nowak2,
Richard Lake1, Willem J. Lesterhuis1
Asbestos Related Diseases. The University of Western Australia,
National Centre for Asbestos Related Diseases. The University
of Western Australia. Sir Charles Gardiner Hospital, Perth, WA,
AUSTRALIA
1
Objectives: Cytotoxic chemotherapeutics form the cornerstone
of treatment for thoracic cancer. Because these drugs typically
demonstrate a clear dose-response relationship, patients are
given the highest dose that does not cause unacceptable side
effects. In contrast, in murine cancer studies chemotherapy
dosages are often based on custom practice from the literature or on small pilot studies for individual drugs, which often
Wayne J. Aston1, Catherine A. Rinaldi1, Scott Fisher1, Anna
Nowak2, Richard Lake1, Willem J. Lesterhuis1
National Centre for Asbestos Related Diseases. The University
of Western Australia. Sir Charles Gardiner Hospital, Perth, WA,
AUSTRALIA
1
Objectives: Identify the dynamics of leucocyte infiltration in
mesothelioma tumours during immunogenic chemotherapy.
Methods: BALB/c mice were inoculated subcutaneously with
5x105 AB1 mesothelioma cells. On d12 the mice were given 250
mg/kg cyclophosphamide intraperitoneally. The tumours were
harvested at various times after chemotherapy and were analysed using H&E histology and flow cytometry to characterise
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the immune cell infiltrates.
Results: AB1 tumours treated with cyclophosphamide regressed, appeared to regrow then regressed completely over a
period of 20 days. Based on this four time points were selected for histology and flow cytometry analysis: growth before
treatment, growth after treatment, the regrowth phase and the
final regression phase. The most notable finding was in the final
regression phase in which the resected tumour section showed
large leukocytic infiltrates. We noted gross changes to immune
cell populations across all time points including a decrease
in FoxP3+ Tregs and a significant increase in both CD4+ and
CD8+ T lymphocytes. Functional studies showed that CD8+ T
cells were necessary for the curative effect of cyclophosphamide.
immune related genes. A central hub in both modules, which
was upregulated in responding mice was inducible nitric oxide
synthase (iNOS). Cotreatment with competitive iNOS inhibitor
L-NNA inhibited the response to anti-CTLA4 while conversely
the NO generator isosorbide dinitrate significantly enhanced
the cure rate from 10 to 80% in AB1-HA mesothelioma-bearing
mice. The drug repurposing approach identified all-trans retinoic acid as a candidate synergistic drug. Indeed, cotreatment
with anti-CTLA4 increased the response rate from 10% to 60%.
Conclusion: This approach allows the identification of therapeutic targets and drugs that act synergistically with checkpoint
blockade in cancer.
Keywords: biomarker, tumour immunology, checkpoint blockade, systems biology
Conclusion: Cyclophosphamide treatment of mesothelioma in
vivo results in leukocytic infiltrates that mediate regression,
particularly CD8+ T cells. Further studies will aim to further
investigate the immunogenic mechanisms behind this.
Keywords: Cyclophosphamide, Immunogenic, chemotherapy
PP01.79: IDENTIFICATION OF REPURPOSED
DRUGS THAT INCREASE IMMUNE CHECKPOINT
BLOCKADE EFFICACY IN MESOTHELIOMA USING
NETWORK ANALYSIS OF RESPONDING TUMOURS
Rachael M. Zemek1, Catherine A. Rinaldi1, Richard Lake2, Michael J. Millward1, Anna Nowak2, Willem J. Lesterhuis2
School of Medicine and Pharmacology, University of Western
Australia, Perth, WA, AUSTRALIA, 2School of Medicine and Pharmacology, National Centre for Asbestos Related Diseases. The
University of Western Australia, Perth, WA, AUSTRALIA
1
Objectives: Immune checkpoint blockade, such as anti-CTLA4
and anti-PD1 has shown impressive results in several cancer
types, with durable complete regression in a proportion of
patients. Conversely, the majority of patients do not display
tumour regression. It is unknown what molecular events
determine this dichotomous response, nor which co-treatments
are likely to combine effectively with checkpoint blockade. We
hypothesized that the effector response in the tumour could be
visualized as a complex network of interacting gene products
and that by mapping this network we could predict effective
therapeutic interventions.
Methods: To compare the molecular events in responders
versus non-responders, we treated mice with bilateral AB1-HA
mesothelioma tumours, which respond symmetrically to anti-CTLA4. We used a weighted gene correlated network analysis
(WGCNA) of gene expression profiling data from responding
versus non-responding tumours to identify modules associated
with response. In addition, we used upstream regulator analysis
and interrogated genome-wide drug-pertubation signatures in
the connectivity map database to identify repurposed drugs
that would phenocopy the response-associated network, and
thus increase the response rate to anti-CTLA4.
Results: We found two modules highly significantly differentially expressed; one enriched for cancer-associated genes, one for
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PP02: POSTER MIXER AND POSTER DISCUSSION SESSION 2
18:00 – 19:30
PP02.02: LOCALIZED SPLENIC REOCCURRENCE IN
MALIGNANT PERITONEAL MESOTHELIOMA
Rivka Thurm, Gleneara E. Bates, Yaakov Bressler, Robert N.
Taub
PP02.01: ART AND SCIENCE: RELATIONSHIP
WITH HUMAN ANATOMY FROM AESTHETIC AND
SCIENTIFIC PERSPECTIVES
Guillermo A. Villamizar
FundClas, Bogotá, COLOMBIA
Objectives: Along art history, different artists were important to get close the nature of anatomy expressed in bones,
muscles and vessels to their audiences through drawings
which permitted to see them in both ways: as science in their
subject and art in their aesthetical potential. To reveal what is
unknown is part art and part science and the body now speaks
itself from deep-seated physical structures of human anatomy,
something the eye of the artist can’t see, neither the physician.
Represent that body is not possible anymore but only using the
new technologies. And this way, it becomes a challenge when
the sensitive spirit wants to show what he can’t see by its own
tools: the eyes. Representation of the human body was for artist
a contribution to the knowledge of diseases, and when this is
not possible anymore, what the artist do is to draw a kind of
social body. Why? It is a representation that only can be done by
mobilizing forces beyond the artist studio and the wisdom that
the drawing can offer to him. This is the reason for artist working now with physicians, scientist, workers and union workers,
lawyers, sociologists, anthropologists and social activists, all
a social body which helps to visualize what the eye can’t see:
that disease which affects not only a person but yes to a wide
population turning out into a problem of public health, as it is
the case of ARD, including mesothelioma.
Methods: This investigation is done from the art field and its
methodologies are not scientific and therefore its methodologies are empirical. This type of job has a symbolic and communicative dimension that is not possible to measure in science
parameters.
Results: During last two years I have been involved –as artistdoing art exhibitions where the main focus of it has been asbestos hazards and ARD too. Every exhibition includes distribution of flyers, presentation of
conferences and production of images alerting to the public in
Colombia about ARD. My activity includes visits to medicine
schools, meetings with union workers and lawyers for discussing actions to prevent and alert about ARD and seeking ways
for banning asbestos in Colombia. Since then, many people in
Colombia are conscious about asbestos hazards and ARD and
Colombian society is giving steps toward banning asbestos.
Conclusion: The mixing of art and science shows the extraordinary possibility that this approach have as a vehicle for
communicate the ARD.
Objectives: Metastases to the spleen, while relatively common
in hematologic cancers, are rare in solid tumors, occurring in
only 0.3–7.3% of cases. The rarity of splenic metastasis in solid
cancers is thought to be a result of the splenic vascular structure inhibiting entry of emboli of metastatic cancer cells. It has
also been proposed that the long retention time (~10 minutes)
of leukocytes within the spleen creates a hostile environment
for tumor seeding and growth. Of splenic metastases, the
majority are found as part of late-stage systemic disease or are
discovered on autopsy.
Methods: An IRB approved, retrospective chart review of mesothelioma patients treated at CUMC from 1995–2015.
Results: A single patient was identified as having an isolated
splenic reoccurrence, which was discovered and monitored by
serial CT scans and confirmed on splenectomy. In 2007, the
68-year-old male presented with weight loss, diffuse mid to
upper body abdominal discomfort, loss of appetite, and postprandial fullness. CT scans showed infiltration of the mesentery
and thickening of the omentum, and he underwent laparoscopy
in June 2007. Several 2 to 4 mm pearly nodules were found on
the parietal peritoneum and gutters bilaterally and the anterior abdominal wall. Pathology showed poorly differentiated
malignant cells consistent with epithelioid mesothelioma. In
August 2007, the patient underwent exploratory laparotomy,
cytoreduction, and HIPEC with Cisplatin. The surgical pathology
report was consistent with MPM with widespread infiltration
of intra-abdominal soft tissue. Histology showed malignant
mesothelioma, biphasic subtype (60% epithelioid, 40% sarcomatoid). The patient then received four cycles of intraperitoneal
(IP) chemotherapy (Doxorubicin/Oxaliplatin) combined with six
cycles of systemic chemotherapy (Gemcitabine/Oxaliplatin), followed by four weekly doses of IP gamma interferon (2 million
units). Follow-up surveillance by CT scan was performed at 3-6
month intervals for three years, then yearly. A routine CT scan
in October 2012 showed a new 33.6 mm mass in the spleen
which mandated a follow-up scan three months later at which
time the mass had grown to 44.6 mm. This prompted surgical
consultation and splenectomy in February 2013. Pathology
showed extensively necrotic malignant mesothelioma, biphasic
subtype (70% epithelioid, 30% sarcomatoid) infiltrating into
the splenic parenchyma. The tumor was WT1 and CK5 positive
and P16 negative, indicating an aggressive phenotype similar
to the parent tumor. Since splenectomy, the patient has been
progression-free.
Conclusion: Quarterly CT scans allowed early detection and
intervention in this case, preventing the disease from spreading
to the peritoneal space. This report demonstrates that continual
monitoring by serial CT scans can help early detection of novel
MPM reoccurrences.
Keywords: mesothelioma, Peritoneal, Spleen, Metastasis
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ponent of histology shows non-sarcomatous type.
PP02.03: RADIOLOGICAL FEATURES OF PLEURAL
MESOTHELIOMA COMPARED WITH CASES
DIAGNOSED BEFORE AND AFTER 2008 IN JAPAN
Keywords: sarcomatous mesothelioma, pleural rind, pleural
effusion without any pleural chages, feature of pleural mesothelioma
Takumi Kishimoto
Reseach Center For Asbesstos-related Diseases, Okayama Rosai
Hospital, Okayama, JAPAN
Objectives: We already presented 7 types of radiological
patterns of pleural mesothelioma(PM) and the pleural rind
occupied 42% and pleural effusion without any pleural changes　12%. In this study, we divided these cases into the time
diagnosed before and after 2008 and compared the percentage
of these patterns.
Methods: Seven hundred and eight cases of pleural mesothelioma who were pathologically confirmed were evaluated. Four
hundred and eighty two cases(group 1) collected from all over
Japan in 2003 to 2008, 90 cases diagnosed before 2008(group
2) and 90 cases after 2009 (group 3) obtained from Okayama
Rosai Hospital. The pattern of chest CT at the time which mesothelioma was diagnosed were classified into 7 category such
as 1)solitary-mass formation, 2)pleural rind, 3)slight thickening
of pleura, 4)thickening of mediastinal pleura, 5)pleural effusion
without any pleural changes, 6)multi-mass formation and 7)
specific type. Furthermore, the radiological patterns of the
histological differences were evaluated.
Results: For histological classification, 364(57.2%) cases
showed epithelioid histology, 112 cases(17.6%) biphasic type
and 155 cases(24.4%) were sarcomatoid type and 5 cases(0.8%) were specific type. The radiological patterns in each
3 groups shows in Figure 1. The pattern of pleural effusion
without any pleural changes occupied 25.4% in group 3. On
the other hand, pleural rind showed only 32.4%. This number
significantly increased than only 7.1 % of group1 and 10% of
group 2. Almost all cases who showed pleural effusion without
any pleural changes were diagnosed in the early stages using
thoracoscopy guided pleural biopsy.　But for sarcomatoid type,
this pattern occupied only 9.1% of Group 3. Fig1
Conclusion: Nowadays, in Okayama Rosai Hospital, about
25% of PM are diagnosed at the early stage which shows only
pleural effusion without any changes of pleural and main com-
PP02.04: CT FINDINGS OF MALIGNANT PLEURAL
MESOTHELIOMA AND CORRELATION WITH THE
SURVIVAL PERIOD
Katsuya Kato1, Kenichi Gemba2, Kazuto Ashizawa3 , Takumi
Kishimoto4 , Nobukazu Fujimoto5, Keisuke Aoe6 , Yukio Takeshima7, Kouki Inai8
Diagnostic Radiology2, Kawasaki Medical School, Okayama,
JAPAN, 2Chugoku-Chuo Hospital, Fukuyama, JAPAN, 3Nagasaki University School of Medicine, Nagasaki, JAPAN, 4Research
Center Of Asbestos-related Diseases, Okayama Rosai Hospital,
Okayama City, JAPAN, 5Okayama Rosai Hospital, Okayama,
JAPAN, 6National Hospital Organization Yamaguchi-Ube Medical
Center, Ube, JAPAN, 7Hiroshima University School of Medicine,
Hiroshima, JAPAN, 8Diagnostic Pathology Center, Hiroshima,
JAPAN
1
Objectives: The aim of this study was to demonstrate CT findings of malignant pleural mesothelioma (MPM) and to determine their correlation with survival time in patients with MPM.
Methods: In total, 142 patients with MPM were continuously
enrolled at Okayama Rosai Hospital from Oct 1955 to Oct
2015. All these patients had 1) pathologically proven MPM; 2)
undergone CT at the time of diagnosis, and 3) follow-up clinical
records. The CT findings at the time of diagnosis were retrospectively reviewed and the survival periods were analyzed. To
assess the extent of MPM lesion, the thorax was divided into
three zones according to the upper border of the aortic arch and
the inflow portion of the inferior pulmonary vein to the heart.
The number of MPM-involved zones was scored as 0–3. We
defined this score as the “extensive score” to quantify the tumor
volume.
Results: We found 142 patients with MPM between 1995 and
2015. Of these, 130 were men and 12 were women. The mean
age at the time of diagnosis was 69 years (range, 42–91 years).
The pathological subtypes were as follows: epithelial type,
99 cases (70%); biphasic type, 16 cases (11%); sarcomatous
type, 25 cases (17%); lymphoepithelial type, 2 cases (1%). The
overall median survival time (MST) was 12.0 months (range,
0.7–103.2 months). Pleural plaques were detected in 62 cases
(44%) and calcification in 46 cases (32%). Asbestosis was
present in only one (1%) case, and no case of diffuse pleural
thickening was found. The MPM-related CT findings were as
follows: circumferential pleural thickening, (“pleural rind”) 43
cases (30%); mediastinal pleural thickening, 104 cases (73%);
interlobar fissural thickening, 78 cases (55%); pericardial
invasion, 42 cases (30%); diaphragmatic invasion, 23 cases
(16%); thoracic volume loss, 72 cases (51%); pleural effusion,
118 cases (83%); pneumothorax, 4 cases (3%); solitary mass
formation, 12 cases (8%); and multiple mass formation, 42
cases (30%). The extensive scores were as follows: 0 points
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ABSTRACT BOOK
(no pleural thickening), 13 cases (9%); 1 point, 16 cases (11%);
2 points, 13 cases (9%); and 4 points, 100 cases (70%). The
median thickness of the thickest part of the MPM lesions was 14
mm (range, 0–87 mm). Lesions with a thickness of 5 mm > (P
= 0.001) and 10 mm > (P = 0.004) and extensive scores of 1–3
(P = 0.013) were significantly correlated with nonepithelial-type
MPM. The other CT findings showed no significant correlation
with the pathological subtypes. On a multivariate analysis, a
low extensive score (0–2 points vs. 3 points) and the epithelial
subtype were associated with longer MST (hazard ratio [HR],
1.95; 95% confidence interval (CI), 1.21–3.04; p = 0.006; and
HR, 2.11; 95% CI, 1.42–3.13; p < 0.001).
Conclusion: Epithelial mesothelioma was significantly associated with thin pleura and a low extensive score at the time of diagnosis. The other CT findings showed no significant correlation
with the pathological subtypes. The epithelial histologic subtype
and a lower extensive score on CT at the time of diagnosis were
significantly related to a longer MST in patients with MPM.
Keywords: CT, survival period, mesotheliom, pleura
PP02.05: ASBESTOS BODY AND PLEURAL
PLAQUE OF PATIENTS WITH MALIGNANT
PLEURAL MESOTHELIOMA WHO UNDERWENT
EXTRAPLEURAL PNEUMONECTOMY
Kazunori Okabe1, Hiroyuki Tao2, Toshiki Tanaka2, Tatsurou
Hayashi2, Kouichi Yoshiyama2, Masashi Furukawa2, Kumiko
Yoshida2, Katsuya Katou3
Division of Thoracic Surgery, Yamaguchi Ube Medical Center,
Ube, JAPAN, 2Yamaguchi Ube Medical Center, Ube, JAPAN, 3
Kawasaki Hospital, Okayama, JAPAN
1
Objectives: Malignant pleural mesothelioma (MPM) has been
recognized as related to asbestos inhalation. Pleural plaque is
the main radiological finding of asbestos inhalation. The aim of
this study is to analyze the asbestos body counts in the lungs
and pleural plaques of patients with MPM who underwent
extrapleural pneumonectomy (EPP).
Methods: Forty consecutive MPM patients who underwent EPP
from June 2006 to August 2014 were reviewed. Asbestos body
quantification involved the digestion of 1-4 grams of lung tissue
in bleach employing a modified Smith and Naylor method1).
Pleural plaque in the contralateral chest was evaluated by CT,
and graded as 0 (none), 1 (low), 2 (moderate), or 3 (high). [Reference] 1) Smith MJ, Naylor B. Am J Clin Pathol 1972; 58:250-254
Results: The median age was 61 years old (42 - 74). 32 males
and 8 females were operated. The median asbestos body count
was 6,540/g dry lung (range: lower than the detection limit 443,571). The asbestos body counts of eight patients (20%)
were less than 1,000/g dry lung. The pleural plaques of 20
patients were graded as 0 (none), 16 patients were graded as 1
(low), 3 patients were graded as 2 (moderate), and 1 patient was
graded as 3 (high). As shown on the table below, the asbestos body count was proportional to the pleural plaque. Table:
Asbestos body count in the lung and pleural plaque grade The
asbestos body counts were listed in numerical order.
asbestos body
/g, dry lung
7,169
7,209
7,706
7,862
7,882
8,125
9,029
9,313
9,471
10,087
16,556
18,756
19,916
24,036
51,073
153,110
159,579
186,649
319,989
443,571
pleural plaque
grade
0
1
0
0
0
0
1
1
1
1
2
0
1
1
2
0
1
3
1
2
asbestos body
/g, dry lung
< lower limit
< lower limit
65
196
265
524
711
878
1,600
1,772
2,030
2,279
2,319
2,340
2,532
2,994
3,412
4,027
5,127
5,911
pleural plaque
grade
0
0
0
0
0
1
0
0
1
1
0
1
0
0
1
1
1
0
0
0
Conclusion: One half of the MPM patients who underwent
EPP had no finding of pleural plaque on CT. The asbestos body
counts in the lungs were less than 1,000/g dry lung in 20%
of the MPM patients who underwent EPP. The asbestos body
count in the lung was proportional to the pleural plaque.
Keywords: pleural plaque, asbestos body, extrapleural pneumonectomy, Malignant pleural mesothelioma
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PP02.06: THE IMAGING JOURNEY OF A PATIENT
WITH MALIGNANT MESOTHELIOMA
Vincent Lam, Jan Brozik, Daniel T. Barnes
2015 demonstrated a large right pleural effusion and cytology
demonstrated epitheloid mesothelioma. He was commenced
on chemotherapy. After 2 cycles, a follow up CT demonstrated
florid peritoneal and pleural disease and he is receiving further
chemotherapy. Radiology Department, Glenfield Hospital, University Hospitals of
Leicester, Leicester, UNITED KINGDOM
Objectives: We will describe and demonstrate, with appropriate examples, the imaging of patients with malignant pleural
mesothelioma (MPM) in a tertiary thoracic centre. We will
highlight salient CT findings relating to:
Diagnosis including appearances post-surgical intervention/
talc pleurodesis.
Pre-operative imaging including contraindications for radical
surgery.
Post-operative appearances, particularly relating to radical versus non-radical surgery, as well as highlighting the difference
between “normal post-operative appearances” and progressive
disease.
Methods: Cases were retrospectively analysed and selected
from the dedicated, regional University Hospitals of Leicester
Mesothelioma Multi-Disciplinary Team (MDT) meeting according to the above criteria, from June 2013 to December 2015.
This MDT evaluated 445 patients (367 male) in this period.
Results: It is important to recognise the normal post-operative
appearances of MPM to correctly guide patient care. Appropriate images exemplifying different aspects of a patient’s imaging
pathway are included. Fig 1. CT Chest Abdomen and Pelvis,
coronal section of a 77 year old male referred for an extended
pleurectomy and decortication with diaphragmatic patch repair.
Post-operative imaging revealed an atypical loculated chylous
collection (denoted by the asterix) with a right basal fluid collection either side of diaphragmatic patch (white arrows).
Conclusion: Imaging is integral to the patient’s journey, beginning from the diagnosis of MPM through to staging, post-surgical follow-up and palliative care. The role of a radiologist is vital
to delivering an optimum patient-centred service.
Keywords: Post-operative appearances
PP02.07: PATTERNS OF DETECTABLE TUMOR
PROGRESSION IN PATIENTS WITH MALIGNANT
PLEURAL MESOTHELIOMA WITH AN FDG-PETNEGATIVE T1A TUMOR
Kozo Kuribayashi1, Takayuki Terada1, Taiichiro Otsuki1, Eisuke
Shibata1, Koji Mikami1, Tohru Tsujimura2, Seiki Hasegawa3 ,
Takashi Nakano1
Hyogo College of Medicine, Department of Respiratory Medicine, Nishinomiya, Hyogo, JAPAN, 2Hyogo College of Medicine,
Department of Pathology, Nishinomiya, Hyogo, JAPAN, 3Hyogo
College of Medicine, Department of Thoracic Surgery, Nishinomiya, Hyogo, JAPAN
1
Fig 2. CT Chest, Abdomen and Pelvis, coronal section. A 78
year old male presented with increased shortness of breath
and previous asbestos exposure. An initial CXR in August
Objectives: T1a refers to a very early tumor that involves only
the parietal pleura of one hemithorax without any mediastinal
or diaphragmatic involvement, and that spares the visceral
pleura. T1 tumors are usually associated with free pleural space
and large effusion. Early in its clinical course, malignant pleural
mesothelioma (MPM) tends to remain localized to the ipsilateral
hemithorax for a long period and radiological tests do not show
any obvious nodules on the parietal pleura. Usually, FDG-PET/
CT cannot detect small malignant pleural nodules. We evaluatiMig2016.ORG
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ABSTRACT BOOK
ed the patterns of detectable tumor progression in patients with
MPM with an FDG-PET-negative T1a tumor.
Methods: Seven MPM patients (5 males, 2 females) with an
FDG-PET-negative T1a tumor were followed-up with FDG-PET at
our institution.
Results: The median interval between the first negative PET
scan and the positive follow-up scan to identify ipsilateral
malignant pleural nodule was 32 months (5-46 months). In two
patients with epithelioid mesothelioma, the median interval,
between the first negative PET scan and positive follow-up
scan to identify malignant nodules at the sites of thoracoscopy,
was shorter at 5 and 8 months, respectively. In 4 patients, PET
detected localized pleural nodules that were 5.1–8.2 mm in
diameter on the CT scan. One patient showed diffuse pleural
thickening with an FDG-PET-positive tumor with a thickness of
6.5 mm.
Conclusion: FDG-PET is a valuable modality for detecting the
progression of T1a tumors >5 mm in diameter. The median
interval between the first negative PET scan and the positive
follow-up scan was long (32 months), and subcutaneous implantation at the site of thoracoscopy is an early manifestation
of MPM.
PP02.08: LYMPHANGITIC CARCINOMATOSIS:
A COMMON FORM OF PROGRESSION IN
FOLLOWING RADICAL PLEURECTOMY
Ian B. Berger1, Charles B. Simone1, Andrew Haas1, Evan Alley1,
Keith A. Cengel1, Joseph Friedberg2, Urooj Khalid1, Akash
Patel1, Sharyn Katz1
Perelman School of Medicine, Philadelphia, PA, UNITED STATES
OF AMERICA, 2University of Maryland School of Medicine, Baltimore, MD, UNITED STATES OF AMERICA
1
Objectives: Lymphangitic carcinomatosis is a pattern of
metastatic progression that is generally associated with a poor
prognosis, but has not been well-described in the setting of malignant pleural mesothelioma. Here, we examine the incidence
of development of lymphangitic carcinomatosis as a pattern of
failure in the setting of radical pleurectomy.
Methods: All patients with a diagnosis of malignant pleural
mesothelioma undergoing radical pleurectomy at our institution
between 2008 and 2012 were included in this retrospective
study. Patients without available post-surgical follow-up CT
imaging for review were excluded. CT images were reviewed
by an experienced, board-certified thoracic radiologist for the
presence of lymphangitic carcinomatosis characterized by
progressive interlobular septal thickening often accompanied
by axial peribronchial thickening. Cases felt to be positive or
potentially positive for lymphangitic carcinomatosis were then
reviewed by consensus by the PENN Mesothelioma and Pleural
Program Multi-disciplinary Team.
Results: Of the patients who underwent radial pleurectomy
in the specified time frame, a total of 46 patients had serial
follow-up imaging available at our institution. A total of 16 of
the 46 patients (34%) developed lymphangitic carcinomatosis
during the post-surgical period. The median time to lymphangitic carcinomatosis was 12 months following surgery, and the
median post-surgical CT follow up period was 19 months.
Conclusion: Lymphangitic carcinomatosis is a common and
likely underreported manifestation of disease progression in patients with malignant pleural mesothelioma undergoing radical
pleurectomy. Additionally investigation is needed to determine
how lymphangitic carcinomatosis impacts survival in patients
undergoing radical pleurectomy and if surgery alters failure
patterns compared with systemic therapy alone in patients with
malignant pleural mesothelioma.
Keywords: pleurectomy, lymphangitic carcinomatosis, CT,
mesothelioma
PP02.09: MALIGNANT PLEURAL MESOTHELIOMA,
DEMOGRAPHIC DATA AND CLINICAL STAGING OF
193 CONSECUTIVE PATIENTS
Amr M. Eldemery1, Abdelrahman M. Abdelrahman2, Fatma
M.A. Aou El-Kasem3 , Rabab M. Gaafar4 , Hisham Wahba5, Maha
Yahia6 , Eman Loay7
Surgery, National Cancer Institute, Cairo, EGYPT, 2Surgery,
National Cancer Institute, Cairo, EGYPT, 3Medical Oncology,
National Cancer institute, Cairo, EGYPT, 4Medical Oncology, National Cancer Institute, Cairo, EGYPT, 5Radiology, National Cancer
Institute, Cairo, EGYPT, 6Medical Oncology, National Cancer
Institute, Cairo, EGYPT, 7Pathology, National Cancer Institute,
Cairo, EGYPT
1
Objectives: Malignant pleural mesothelioma is a rising health
problem in some countries allover the world including ours. In
spite of rarity of this disease , we have rising incidence every
year and the curve of referral to our institution is going up.
The aim of this study is to outline the demographic data of our
patients, analyze the clinical staging based on radiology and its
relation to pathologic subtype of the disease.
Methods: This is a retrospective study included 193 patient
with pathology proven pleural mesothelioma referred to our
hospital during the period from May 2014 to August 2015 .
Computed tomography scan of the chest and abdomen is the
primary diagnostic and staging modality in all patients. Full history Taking including residence near by or working in asbestos
related industry. All radiologic data were revised with the same
radiologist .
Results: Of the 193 patients studied, we have 96 males with
male to female ratio 1:1, the mean age was 43years. Right sided
disease was present in 63.7% , bilateral disease was found in
only 3 patients at the time of diagnosis. Pleural effusion was
the only radiologic finding in 45.6% ,diffuse pleural thickening
was found in 24.4%, 15.5% had nodular thickening , 9.8% had
combined thickening and effusion and only 4.7% presented
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rax by CT was found in 37.8%, mediastinal fat invasion was
found in 36.8%, chest wall invasion in 9.3%, 4% had suspected pericardial invasion 2 of them with pericardial effusion.
Adjacent organ invasion was found in 10 patients (5.1%). Lung
invasion was found in 37 patients and there were 6 patients
with metastatic disease at presentation ( 2 liver, 2 bone , 1 brain
and one suprarenal) . Enlarged mediastinal lymph nodes by CT
was found in72 patients (37.3%). By Computed tomography,
45% of our patients were stag 1, 33.3% stage III, 12.4% were
stage IV and only 9.3 were stage II. Some patient were referred
after pathologic diagnosis, for the rest of undiagnosed patients
, biopsy either CT guided , open or thoracoscopic biopsy was
done. There were 172 with epithelial histology, 11 with biphasic,
6 with sarcomatoid histoogy and we have 4 patients with pathology proven adenocarcinoma without any evidence of other
primary. Two patients with biphasic histology had mediastinal
nodal enlargement by CT, 2 with pulmonary nodules, one with
both nodal enlargement and chest wall invasion and one with
pericardial effusion. Of the 6 patients with sarcomatoid histology , one had metastatic disease and 2 had T2 ( pulmonary
nodules) disease.
Conclusion: Malignant pleural mesothelioma is arising health
problem. Pleura effusion is a good clinical sign that lead to early
symptomatology and diagnosis at early stage. Patients should
be evaluated and treated in highly specialized centers. Patients
with biphasic and sarcomatoid histology usually present with
late stage.
Keywords: Mesothelioma, clinical, staging
PP02.10: INTO THE DEEP: CLOSER LOOK AT
IMMUNE CELLS AND IMMUNE CHECKPOINT
EXPRESSION IN HUMAN MALIGNANT PLEURAL
MESOTHELIOMA
Elly Marcq1, Vasiliki Siozopoulou2, Jorrit De Waele1, Jonas Van
Audenaerde1, Karen Zwaenepoel2, Christophe Hermans1, Niel
Hens3 , Patrick Pauwels2, Jan Van Meerbeeck4 , Evelien Smits1
Center for Oncological Research, University of Antwerp, Antwerp,
BELGIUM, 2Department of Pathology, Antwerp University Hospital, Antwerp, BELGIUM, 3Interuniversity Institute for Biostatistics
and statistical Bioinformatics, Hasselt University, Diepenbeek,
BELGIUM, 4Thoracic Oncology/MOCA, Antwerp University Hospital, Antwerp, BELGIUM
1
immune cells in the tumor microenvironment.
Methods: Immunohistochemistry was used to examine the
expression of several immune cell markers, as well as the
expression of PD-1 and PD-L1, in formalin fixed paraffin embedded (FFPE) tissue of 54 MPM patients (42 at diagnosis, 12
treated with chemotherapy). Identification of different subsets
of cells present in MPM fluids (ascites and pleura) was done
using multicolor flow cytometry and an enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of
soluble PD-L1 in 10 MPM serum samples. To examine whether
correlations exist between the expression data and several
clinicopathological parameters of the MPM patients statistical
analysis is being performed.
Results: Immunohistochemistry showed PD-1 expression on
lymphocytes in 67% of the treated and untreated samples,
while the expression on tumor cells was only found in few
samples. PD-L1 expression on its turn was seen on lymphocytes and on tumor cells, for the latter only in untreated tissues.
All samples showed CD45RO positive lymphocytes. CD4+ and
CD8+ lymphocytes were found in the stroma and in hotspots
of the lymphoid aggregates in all samples. In more than half
of the untreated samples, a subset of the CD4+ cells was also
FoxP3+. The same holds true for the treated samples. Compared to the untreated, more CD4+FoxP3+ cells were found
in lymphoid aggregates of the treated samples (36% vs 50%).
Stromal CD68+ histiocytes and macrophages were found in all
tissue samples. TIM-3 expression was found on tumor cells and
lymphocytes in untreated, as well as in treated samples. Flow
cytometry showed PD-1 expression on CD3+CD4+ T cells and
CD3-CD56+ natural killer cells. PD-L1 was expressed on the
CD11c+CD303+ dendritic cells. Correlations between expression data and clinicopathological parameters are currently
being analyzed and will be presented. Soluble PD-L1 was found
in all serum samples, with a concentration ranging from 0.71
ng/ml till 2.33 ng/ml.
Conclusion: Immunohistochemistry and flow cytometry
revealed the diversity of immune cells present in MPM. Since
some of those cells express PD-1 or PD-L1, it would be worth
further investigating the effect of immune checkpoint blockade
in MPM. Reactivating immune responses that are silenced by
immune checkpoint receptor-ligand interaction might offer new
opportunities for the improvement of therapeutic strategies for
mesothelioma. Surprisingly, all patient serum samples were
positive for soluble PD-L1, requiring further investigation to
determine its value as biomarker.
Keywords: immune checkpoints, tumor microenvironment,
Immunotherapy
Objectives: Immune checkpoints, such as programmed
death-1 (PD-1), are responsible for controlling and inactivating
the immune system in order to avoid autoimmunity and prevent
tissue damage. Blocking the ligand-immune checkpoint interaction already showed promising results in several cancer types.
Data derived from a small number of mesothelioma patients
suggest that blocking immune checkpoints could offer new opportunities for treatment of this very aggressive tumor. Gaining
more insight in the immunological aspect of the tumor microenvironment in human malignant pleural mesothelioma (MPM)
would be of great interest to develop an efficient immunotherapy. Therefore, we investigated the expression of PD-1 and its
ligand PD-L1 in human MPM and identified different subsets of
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PP02.11: DEVELOPMENT OF NOVEL
THERAPEUTICS TARGETING MALIGNANT PLEURAL
MESOTHELIOMA
PP02.12: INHIBITION OF METHYLTRANSFERASE
EZH2 IMPROVES TUMORICIDAL ACTIVITY OF
MACROPHAGES AGAINST MESOTHELIOMA CELLS
Perry Devo
Malik Hamaidia, Luc Willems
Pharmaceutical, Environmental And Chemical Sciences,
University of Greenwich, London, UNITED KINGDOMObjec-
Molecular And Cellular Epigenetics, Interdisciplinary Cluster for
Applied Genoproteomics (GIGA) of University of Liège, Liège,
BELGIUM
tives: · To synthesise and investigate the anticancer potential of
novel and newly discovered natural products against malignant
pleural mesothelioma (MPM). · To synthesise and evaluate a
number of analogues both to investigate the mode of action of
these compounds and to improve on their anticancer potency. Methods: Natural products JBIR23, -24, -31, -88, -101 and -102
have all been isolated from various species of Streptomyces.
Each compound has been shown to have anti-cancer properties against the ACC-MESO-1 cell line. Fragments building up
to the natural product were also tested against MPM cell lines
to evaluate their structure activity relationship and analogues
were created depending on their anticancer activity.
Results: Various attempts towards the synthesis of JBIR-23 and
JBIR-101 will be reported. Using an optimised synthetic route,
a number of chemical analogues have also been synthesised.
These synthetic analogues were assessed for their anticancer
activity and further analogues were synthesised to improve on
their potency. The biological activity against the ACC-MESO-1
cell line will be presented.
Conclusion: MPM is an aggressive neoplasm and current
therapeutics are ineffective in the treatment of this malignancy.
Therefore the investigation and development of potential novel
chemotherapeutics is essential going forward. Our research to
date has shown that natural products are a valuable source for
the identification of novel compounds in the future treatment of
MPM.
Keywords: medicinal chemistry, Preclinical, chemotherapy,
natural products
Objectives: Clinical evidence indicates that tumor infiltration
by tumor associated macrophages (TAMs) correlates with poor
prognosis in malignant mesothelioma (MM). By attenuating
the immune response, TAMs indeed promote survival of MM
cells. TAMs share properties with alternative macrophages (M2)
and are activated by anti-inflammatory (e.g. IL-10) or Th2-associated (i.e. IL-4, IL-13) cytokines. In contrast, classical (M1)
macrophages are stimulated by interferon (IFN)-γ and microbial
components (e.g. LPS). We hypothesized that macrophage
activation is mediated by a transcriptional program tightly regulated by epigenetic modifications. We focused on the Polycomb
Repressive Complex 2 (PRC-2) EZH2 lysine methyltransferase responsible for trimethylation of histone H3 at lysine 27
(H3K27me3). We investigated the effect of a selective EZH2
inhibitor (EPZ005687) on tumoricidal activity of Raw 264.7 and
primary human monocytes-derived macrophages.
Methods: Raw 264.7 macrophages were cultivated in presence
of EPZ005687 for 24h and then LPS for 24h. Human macrophages were treated with EPZ005687 for 24h before polarisation into M1 (in presence of LPS and IFN-γ) or M2 (with IL-4).
The phagocytic activity was evaluated by using dextran-FITC.
ROS production was determined with the DCFH-DA probe.
Expression of CD206 and HLA-DR was analysed by flow cytometry. Cytotoxic activity was performed by incubating AB-1 and
M14K mesothelioma cells in media supplemented with 25%
or 50% of macrophage supernatant. Viability of mesothelioma
cells was assessed by performing MTS assay and Annexin-V
labeling. NADPH oxidase was inhibited with apocynin.
Results: Our data show that inhibition of EZH2 increases
phagocytosis in response to LPS stimulation, reduces CD206
expression by human M2 macrophages and enhances cytotoxic
activity against mesothelioma cells. We further demonstrate
that EZH2 inhibition stimulates macrophage killing activity via
reactive oxigen species produced by NADPH oxidase.
Conclusion: The inhibition of EZH2 enhances the tumoricidal
potential of macrophages. This strategy could improve immunotherapy of MM patients.
Keywords: Immunotherapy, epigenetic inhibitors, Malignant
mesothelioma, Tumor associated macrophages
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ABSTRACT BOOK
PP02.13: THE DL 922-947 ONCOLYTIC VIRUS AS
A NEW POSSIBLE THERAPEUTIC TOOL AGAINST
MESOTHELIOMA
Carmelina A. Iannuzzi1, Carmela Passaro2, Iris M. Forte3 , Paola
Indovina4 , Giuseppe Portella5, Antonio Giordano6 , Francesca
Pentimalli3
Department of Medicine, Surgery And Neuroscience, University
of Siena, Siena, ITALY, 2Dipartimento Di Scienze Mediche Traslazionali, University of Naples, Naples, ITALY, 3Oncology Research
Center Mercogliano (crom), National Cancer Institute of Naples
Pascale Foundation, Avellino, ITALY, 4Sbarro Institute for Cancer
Research and Molecular Medicine, Philadelphia, PA, UNITED
STATES OF AMERICA, 5University of Naples, Naples, ITALY, 6University of Siena, Siena, ITALY
1
Objectives: Malignant mesothelioma (MM) is a highly aggressive cancer for which, at present, no curative modalities
exist. MM in fact is poorly responsive to the current therapeutic
strategies, resulting in a dismal prognosis. New alternative
therapeutic approaches are therefore urgently needed. Here
we aimed at investigating at the preclinical level whether
oncolytic viruses could represent a feasible strategy. Oncolytic
viruses have different advantages as anticancer agents: they
replicate selectively in tumor cells with amplification of the
input dose; they are able to stimulate the antitumoral immune
response; they can be used in combination with other cytotoxic
agents; the pleural cavity in which pleural MM arises is easily
accessible for such therapeutic approach. We focused on the
dl 922-947 adenovirus having a 24 bp deletion in the E1A-conserved region 2, which binds and inactivates the retinoblastoma
protein, resulting in a virus that cannot trigger S phase entry
in normal cells, but can still replicate in cells with an aberrant
G1-S checkpoint, a defect observed in over 90% human cancers, including MM.
Methods: We studied on NCI-H28, NCI-H2452, MSTO-211H
and NCI-H2052 (a panel of mesothelioma cell lines representative of the main different histotypes) the effects of dl 922-947
treatment used both as a single agent or in combination with
other strategies (cisplatin and MK-1775). We analyzed the
effects of these treatments on cell viability through sulforhodamine B (SRB) assay and FACS analyses. We analyzed the
synergy among various treatment combinations through the
Calcusyn Software and by Western blot we evaluated the underlying molecular mechanisms.
Results: At first, dl 922-947 cytotoxicity was evaluated through
SRB, which showed that all MM cell lines were susceptible to
viral treatment, except NCI-H2052 cells, in which viral entry
was not efficient, as shown through infection with a reporter
adenovirus transducing GFP. Interestingly and consistently
with the cytotoxic effect observed, FACS analysis showed that
dl 922-947 treatment induced an increase of the subG1 cell
fraction (suggestive of apoptosis induction) and of the hyperdiploid (4N) population (suggestive of mitotic defects). We also investigated by SRB the possible cytotoxic effects of dl922-947 in
combination with other therapeutic strategies. In particular, we
analyzed the effect of dl922-947 in combination with cisplatin,
which is the first-line treatment against MM, and found that, by
comparing different schedules of treatment, cisplatin increased
the cytotoxic effect of the oncolytic virus. We also tested dl922947 efficacy in combination with MK-1775, an efficient inhibitor
of the WEE1 kinase, which is currently being tested in clinical
trials. We found that MK-1775, at doses equal to or above its
IC50 value, is able to increase the cytotoxic effect of the oncolytic virus treatment.
Conclusion: In conclusion our data indicate that treatment
with the dl922-947 oncolytic virus might be a promising new
approach against mesothelioma and warrants further investigation both as single agent and in combination strategies.
Keywords: Oncolytic viruses, WEE1 kinase inhibitor, combination therapy, mesothelioma
PP02.14: CHARACTERISING CTL RESPONSES
AGAINST MESOTHELIOMA NEO-ANTIGENS
Jonathan Chee, Shaokang Ma, Jenette Creaney, Bruce
Robinson
School of Medicine and Pharmacology, National Centre of Asbestos Related Diseases, University of Western Australia, Nedlands,
WA, AUSTRALIA
Objectives: A feature of carcinogen-induced cancers is the
accumulation of mutations in the cancer cell. High throughput
sequencing such as RNA and exome sequencing has allowed
us to interrogate mutations from different cancers. Application
of algorithms on sequencing data allows us to predict mutated proteins in a cancer that can be potentially recognised by
host immune cells such as cytotoxic CD8 T cells (CTLs). CTLs
respond against mutated proteins (neo-antigens), and in some
experimental models, boosting CTL responses specifically
against neo-antigens can cause tumour regression. We have
sequenced mouse mesothelioma (AB1/AB1-HA), and described
the first mesothelioma neo-antigens. Furthermore, we have
demonstrated immune responses against one of the predicted
neo-antigens: mutated Uqcrc2 (Uq2) in mesothelioma bearing
animals. The objective of this study was to characterize further
characterize T cell responses against neo-antigens in tumour
bearing animals after different treatments that cause tumour
regression in AB1 model. We hypothesize that treatments will
increase the strength of responses to ‘existing’ neo-antigens
such as Uq2, and could also lead to the detection of responses
against other neo-antigens that would otherwise be undetectable without treatment.
Methods: Mice were inoculated subcutaneously with mesothelioma cell lines (AB1-HA). AB1-HA bearing animals were
treated either with anti-CTLA4 antibody (checkpoint blockade immunotherapy), anti-GITR antibody(Treg modulation),
gemcitabine (chemotherapy) or depleted of Tregs (removal of
immunosuppression). We tested T cell responses at the draining
lymph nodes and tumours against predicted neo-antigens using
different immunoassays such as IFNg ELISPOT, pMHC tetramers and flow cytometry.
Results: The frequency of T cells specific for a single neo-antigen (Uq2) is low (<0.1%). ELISPOT was the only immunoassay
tested that could detect responses against neo-antigens. Conventional pMHC staining and flow cytometry was not sensitive
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enough to detect a positive signal for low frequency T cells.
Using the ELISPOT assay, we demonstrate the frequency of T
cell responses to neo-antigen Uq2 was highly variable between
mice after treatment, and showed a general trend of increase
after checkpoint blockade immunotherapy, chemotherapy or
Treg removal (compared to non-treated mice). Treatments did
not increase T cell responses to 30 other predicted neo-antigens.
Conclusion: As the frequencies of neo-antigen specific T cells
are low, we were only able to consistently detect neo-antigen
responses with one immunoassay. Assays with increased sensitivities, or an additional T cell expansion step must be used to
detect and phenotype neo-antigen specific T cells. Even though
we detected an increase in neo-antigen T cell responses after
treatment, further work needs to be done to validate if Uq2 or
other neo-antigen responses are essential for an anti-tumour
immune response.
Keywords: Neo-antigens, Next generation sequencing, Immunotherapy
PP02.15: GENE PROFILE OF MESOTHELIOMA
TUMORIGENESIS AND INITIATION AFTER
CHEMORADIATION TREATMENT IN VITRO AND IN
VIVO MODELS
to observe tumorigenesis.
Results: Prime-PCR results indicated that some genes (Bcl2,
Birc5, Bub1, Ccnb1, Gjb6, Foxa1, Muc1, Ndc80, Nkx2-1, Sox2)
are upregulated more than 10 times in the peritoneum of mice
after RN5 tumor challenge compared with naïve mice. Treatment with chemo- or radiation therapy resulted in increased
expression of identified genes of interest in both RN5 and AB12
cell lines. Despite the presence of a lower number of cells
(105 and 104) among the chemoradiation-treated surviving cells,
their implantation resulted in similar tumor incidence and 10
times higher tumor growth than the use of a higher number of
number of parental cells (106 and 105), suggesting that surviving
tumor cells after chemoradiation share the property of stemness. RT-PCR results demonstrated that some of the genes associated with stemness (CD44, Sox2, Sca-1, Birc5) had already
been demonstrated in mesothelioma and other types of cancer.
However, other genes (Batf, Foxa1, Ndc80, Wwc1, Nkx2-1, etc)
had not been reported in mesothelioma stem cells. The potential importance of these newly detected genes was confirmed
by confirming their presence in highly enriched mesothelioma
stem cells as positive controls. Further functional characterization of these genes and their potential role to target stem cells
with immunotherapy needs to be studied.
Conclusion: Molecular identification of mesothelioma cells
and stem cells may provide a novel venue for immunotherapy
against mesothelioma cells as well as cancer stem cells.
Keywords: Murine mesothelioma, cancer stem cell, primePCR, Immunotherapy
Licun Wu1, Zhihong Yun1, Walter Blum2, Beat Schwaller2,
Emanuela Felley-Bosco3 , Marc De Perrot1
Thoracic Surgery, Toronto General Hospital and Princess
Margaret Cancer Center, Toronto, ON, CANADA, 2Department of
Medicine and Anatomy, Department of Medicine and Anatomy,
University of Fribourg, Fribourg, SWITZERLAND, 3Department of
Molecular Oncology, University Hospital Zurich, Zurich, SWITZERLAND
1
Objectives: Immunotherapy has shown promising results for
cancer treatment including mesothelioma, however, the major
issue is a lack of specific antigens for monitoring the immune
response and to target tumor stem cell specifically. To overcome this hurdle we attempt to look for potential candidate
genes determining tumor cell initiation and proliferation in
mesothelioma. Based on this finding, it would then potentially
be possible to design specific immunotherapy to target mesothelioma stem cells.
Methods: We employed the newly developed Prime-PCR
assay specifically designed for mouse mesothelioma to screen
the genes of interest. The expression of the selected genes
was confirmed by real time-PCR, flow cytometry and immune
fluorescent staining. Murine mesothelioma cells RN5 and
AB12 were both developed by intraperitoneal (ip) injection of
asbestos fibre. RN5 cells were injected ip into mice and the
peritoneum tissues were collected at different time points to
determine gene expression. Both cell lines were treated with
cisplatin-based chemotherapy or Cs37 γ-ray radiation in vitro to
evaluate the expression of these selected genes using real-time
PCR and flow cytometry. Serial numbers of surviving tumor
cells after chemo- or radiation therapy were injected into mice
PP02.16: THE ELEVATED LEVELS OF G-MDSC
IN MESOTHELIOMA PATIENTS INHIBIT T CELL
PROLIFERATION AND FUNCTION BY ROS
GENERATION
Swati Khanna1, Francis Mussai2, Anish Thomas1, Gary
Middleton2, Constance Yuan3 , Betsy Morrow1, Jingli Zhang1,
Ira Pastan4 , Maryalice Stetler-Stevenson3 , Raffit Hassan1
Thoracic and Gastrointestinal Oncology Branch, National Cancer
Institute, Bethesda, MD, UNITED STATES OF AMERICA, 2Institute
of Immunology and Immunotherapy, University of Birmingham,
Birmingham, UNITED KINGDOM, 3Laboratory of Pathology,
National Cancer Institute, Bethesda, MD, UNITED STATES OF
AMERICA, 4Laboratory of Molecular Biology, National Cancer
Institute, Bethesda, MD, UNITED STATES OF AMERICA
1
Objectives: The role of myeloid derived suppressor cells
(MDSCs) in mediating tumor immunosuppression in patients
with malignant mesothelioma has not been well characterized.
The goal of our study was to analyze for the presence of both
monocytic and granulocytic myeloid cells in peripheral blood
of mesothelioma patients, evaluate their immunosuppressive
capability and characterize their mechanism of suppression of
T cell responses.
Methods: We evaluated the peripheral blood of patients with
mesothelioma (n=25) and healthy donors (n=20) for the presiMig2016.ORG
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ABSTRACT BOOK
ence of granulocytic myeloid cells (CD11b+CD15+CD14-HLADR-)
and monocytic myeloid cells (CD11b+CD14+HLADR-) by a flow
cytometry panel. Due to higher frequency of granulocytic
myeloid cells, the T cell suppression assays were setup using
sorted autologous T cells and granulocytic myeloid cells. T cell
proliferation and function was evaluated by CFSE dilution and
IFN-γ released by T cells in the supernatants, respectively. To
determine the mechanism by which Gr-MDSC inhibit T cells, we
tested the levels of reactive oxygen species (ROS), and nitrite
oxide species and arginase activity in Gr-MDSC (with or without
inhibitors) sorted from peripheral blood using flow cytometric
and colorimetric assays. Arginase inhibitor nor-NOHA and iNOS
(inducible nitric oxide synthase) inhibitor L-NMMA were used at
a concentration of 500 μM. TBHP was used as a positive control
for ROS assay while D-NMMA was used as a negative control
for nitrite assay.
Results: The granulocytic myeloid cells were significantly
elevated in peripheral blood of mesothelioma patients as
compared to healthy donors (61.2% vs. 47.8%; p=0.009). The
levels of monocytic myeloid cells in patients and healthy donors
were 0.2% and 0.1%, respectively (p=0.04) and were much
lower than the granulocytic subset. Granulocytic subset was
investigated further due to the their elevated levels and higher
frequency in mesothelioma patients. Granulocytic myeloid cells
from mesothelioma patients were found to inhibit the proliferation of both autologous CD4 and CD8 T cells (mean inhibition
of proliferation of 61.9% for CD4 and 75.5% for CD8 at 1:2
T: G-myeloid ratio), which was accompanied with ~ 10 fold
decrease in IFN-γ levels in co-culture supernatants. Thus, this
granulocytic subset is in fact granulocytic myeloid derived suppressor cell (G-MDSC). The arginase levels in G-MDSC (0.3±0.3
vs. 0.2±0.3 μM) and nitrite levels in G-MDSC culture supernatants (1.8±2.5 vs. 0 μM) were below detection limit in both
patients and normal donors and were marginally affected by the
addition of their respective inhibitors (nor-NOHA and L-NMMA). The addition of these inhibitors also did not reverse the
suppressive effect of G-MDSC on T cells proliferation. However,
the average levels of ROS in G-MDSC derived from mesothelioma patients were 3 folds higher than that from healthy donors
(average MFI ~15000 vs. ~5000; p=0.03).
PP02.17: NOVEL MEDICINAL CHEMISTRY
APPROACHES IN MESOTHELIOMA: THE ROLE OF
NATURAL PRODUCTS
Adrian Dobbs
Pharmaceutical, Chemical and Environmental Sciences, University of Greenwich, Chatham Maritime, Kent, UNITED KINGDOM
Objectives: Human malignant pleural mesothelioma (MPM)
is a rare and aggressive neoplasm that originates in the pleura
and is highly invasive. It is generally associated with exposure
to asbestos fibres. One of the greatest challenges is that it is
resistant to most conventional therapies, including chemotherapy, radiotherapy and surgery and the prognosis for patients is
poor. This makes the development of new therapeutic agents all
the more crucial. Historically, the majority of drugs on the market – for any symptom – have their origins in a natural product
(something isolated from nature). Unfortunately there are no
novel drugs in development specifically targeting MPM: principally because until very recently, there were no reported natural
products to serve as the lead compound. This changed in 2009,
with the report of JBIR-23 and -24, two microbial metabolites
isolated fromStreptomyces sp. AK-AB27 : the first natural products
specifically to demonstrate cytotoxicity against MPM cell lines,
with modest IC50 values. More importantly, JBIR-23 inhibited
the proliferation of MPM cells that were otherwise resistant to
clinical anticancer drugs and without evident side effects in
mice. Since the report of JBIR-23, a further 6 compounds have
been reported, also demonstrating activity against ACC-MESO-1 cells with similar IC50 values.
(See next page.)
Conclusion: In summary our results show that G-MDSC are a
major immunosuppressive cell population in mesothelioma patients and they suppress T cell proliferation and function mainly
through generation of reactive oxygen species.
Keywords: mesothelioma, Myeloid derived suppressor cells,
ROS, Arginase
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ABSTRACT BOOK
However, no synthetic or medicinal chemistry studies towards
the preparation of these compounds, or any structure-activity
studies based around them as lead compounds has been conducted. Thus the aim of work is to investigate the synthesis and
mode of action of these compounds.
Methods: We have employed the tools of synthetic chemistry
in developing syntheses of these natural products. These have
been designed to be highly modular and flexible, to permit
rapid preparation of analogues of the natural products. We shall
report the synthetic methodologies developed and the various
libraries of compounds that have been prepared. Biological
data will be reported against ACC-MESO-1 cell lines.
Results: We shall report our studies on the synthesis and
medicinal chemistry of these lead compounds: our successful
attempts at their preparation and also the construction of related analogues; we shall also report initial biological data relating
to these libraries of compounds
Conclusion: There is an urgent and unmet need for new
approaches and novel agents to treat MPM. Exploiting natural
products via synthesis, analogue preparation and structure-activity studies is one such approach. The results reported herein
have shown that this traditional medicinal chemistry approach
towards drug discovery has considerable potential in finding
potential new therapies for MPM.
Keywords: medicinal chemistry, organic chemistry, natural
products
PP02.18: HMGB-1 RELEASE AND THE CD8+ T CELL
RESPONSE ELICITED BY RADIATION TREATMENT
Matthew Wu1, Luis D.L. Maza1, Licun Wu1, Holly Guo1, Hana
Yun1, Marc De Perrot2
Thoracic Surgery, University Health Network, Toronto, ON,
CANADA, 2Thoracic Surgery, Toronto General Hospital and
Princess Margaret Cancer Center, Toronto, CANADA
1
Objectives: Surgery for Mesothelioma After Radiation Therapy
(SMART) has demonstrated substantial benefits in patient
survival compared to other treatment modalities. The benefits
exhibited by SMART treatment may rely on immune activating
effects of radiation before surgery through the release of pro-inflammatory molecules, such as Danger Associated Molecular
Pattern (DAMP) molecules associated with cancer cell death.
We studied the role of HMGB-1 released from dying tumor cells
as an immune system potentiator after radiation treatment for
Malignant Pleural Mesothelioma (MPM).
Methods: Human and mouse epithelioid MPM cell lines were
irradiated in a single fraction of 25Gy radiation. Cell death was
measured by Annexin V and eFluor450 viability dye staining.
Immunohistochemical staining of HMGB-1 was compared
between paraffin embedded MPM samples from patients who
received no treatment (biopsy or extrapleural pneumonectomy
(EPP) ) or Surgery for Mesothelioma After Radiation Therapy
(SMART) treatment. Mice were inoculated with the mouse epithelioid MPM cell line AE-17-OVA and were split into 1) untreated 2) radiation treated and 3) radiation treatment + HMGB-1
blockade groups. Radiation treatment was delivered in three
5Gy fractions of radiation (15Gy total dose) on days 8, 9, and 10
after cell injection. Serum HMGB-1 was measured by ELISA and
tissue staining was performed by immunofluorescence.
Results: In vitro radiated epithelioid MPM cell lines demonstrated increased HMGB-1 release that correlated with
increased viability dye staining. We did not find any significant
difference in staining intensity of HMGB-1 stained MPM paraffin
embedded patient samples. Radiation treated mice showed
slower tumor growth, increased serum HMGB-1, and increased
tumor infiltrating CD8+ T Cells. Blocking of HMGB-1 during
radiation led to increased tumor growth compared to radiation
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ABSTRACT BOOK
LRT in combination with HMGB-1 demonstrates greater tumor
growth compared with LRT alone. AE-17-OVA mice either
received no treatment (N=5), local radiation therapy (N=4), or
local radiation therapy in combination with anti-HMGB-1
antibody (N=5, 100µg per day on days 8-12 after tumor inoculation). Untreated mice (NoRx) demonstrate the greatest tumor
growth compared to LRT + anti-HMGB-1 and LRT groups. Mice
receiving LRT + anti-HMGB-1 exhibit greater tumor volume than
mice treated with LRT alone.
Conclusion: This work supports the beneficial pro-inflammatory role of HMGB-1 in MPM treatment. Thus future studies of
HMGB-1 as a therapeutic strategy would be valuable.
Keywords: HMGB-1, Immunotherapy
immune suppressive molecule PD-L1, costimulation molecules
B7-1/2, MHC class II, macrophage markers, cancer stem cell
markers and so on, which are potentially applied to design
specific immunotherapy against mesothelioma. Tumor-specific
immunity against RN5 mesothelioma was evaluated by in vitro
cell killing assay. Splenocytes derived from RN5-bearing or
naïve mice, and RN5 cells were used as effectors and targets,
respectively. After overnight co-culture of effector T cells (E)
and RN5 cells as targets (T) with pulsation of whole cell lysate,
both effectors and targets were harvested to determine T cell
activation and RN5 cell proliferation.
Results: RN5 cells express a wide variety of immune associated phenotypes PD-L1, B7-1/2, Gr-1, MHC II and D11c, and
mesothelial precursor markers (mesothelin/CD34/CD90),
and also express cancer stem cell markers (CD44, Sox2,
Oct4, Sca-1) at different levels. Interestingly, they share some
macrophage phenotypes MHC II, CD11b, F4/80, CD68, CD206
and Arg-1. Our results indicated that splenocytes derived from
RN5-bearing mice were able to be activated by pulsation of
whole tumor cell lysate, thus resulting in cytotoxic lysis of target
cells. RN5 cell proliferation was significantly suppressed by
activated T cells in an E:T ratio dependent manner. CD8 T cell
activation and proliferation was 2-3 times higher after pulsation
with tumor cell lysate than that in the non-pulsed group. On
the contrary, tumor cell proliferation rate was 30.6±4.4% and
17.6±3.0% at E:T10 and E:T50, respectively, significantly lower
compared with target cells alone (87.5±0.5%).
Conclusion: Tumor-specific immunity can be generated by
pulsation with whole cell lysate. We need to explore the potential antigens that are involved in this immune response.
Keywords: tumor cell lysate, Immunotherapy, Murine mesothelioma RN5, phenotype
PP02.19: IMMUNOPHENOTYPE OF A NOVEL
MURINE MESOTHELIOMA CELL LINE RN5 AND
SPECIFIC IMMUNITY GENERATED BY PULSATION
WITH CELL LYSATE
Licun Wu1, Zhihong Yun1, Walter Blum2, Beat Schwaller2,
Emanuela Felley-Bosco3 , Marc De Perrot1
Thoracic Surgery, Toronto General Hospital and Princess
Margaret Cancer Center, Toronto, ON, CANADA, 2Department of
Medicine and Anatomy, Department of Medicine and Anatomy,
University of Fribourg, Fribourg, SWITZERLAND, 3Department of
Molecular Oncology, University Hospital Zurich, Zurich, SWITZERLAND
1
Objectives: This newly generated murine mesothelioma model
was reported recently by Blum et al from our team. RN5 cell
line was established from neurofibromatosis 2 (merlin) heterozygous (Nf2+/-) mice, as evidence has indicated that approximately half of the malignant pleural mesothelioma patients
have Nf2 mutation. Characterization of the novel mesothelioma
cell phenotypes needs to be investigated in order to develop
personalized immunotherapy.
Methods: Cultured RN5 cells were used to determine the
phenotypes that are associated with immune response, such as
PP02.21: FOCAL ADHESION KINASE INHIBITION
TARGETS MACROPHAGES IN VITRO AND IN VIVO
IN A MESOTHELIOMA MOUSE MODEL
Lysanne Lievense1, Floris Dammeijer1, Menno Van Nimwegen1,
Koen Bezemer1, Yan Wang2, Jonathan Pachter2, Joost
Hegmans1, Joachim Aerts1
Pulmonary Medicine, Erasmus MC Cancer Institute, Rotterdam,
NETHERLANDS, 2Verastem Inc., Boston, MA, UNITED STATES OF
AMERICA
1
Objectives: Recent evidence suggests that FAK inhibition
could have a beneficial effect on the anti-tumor immune
response via depletion of regulatory T cells and increase in
cytotoxic T cells in addition to targeting cancer stem cells. This
leads to new immunotherapeutic possibilities for FAK inhibitors
despite the recent negative results of the COMMAND trial in
advanced mesothelioma patients which used a FAK inhibitor,
VS-6063, as single agent maintenance after chemotherapy
doublet treatment. Tumor-associated macrophages (TAMs) of
the M2 phenotype are prominent immunosuppressive cells in
the mesothelioma microenvironment and a promising therapeutic target, especially in combination with checkpoint inhibitors
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ABSTRACT BOOK
and cellular immunotherapies. Since FAK is known to regulate
diverse macrophage functions, FAK inhibition could be a strategy to target TAMs. The aim of this study is to investigate the
influence of FAK/PYK2 inhibition on macrophage phenotype in
vitro and macrophage tumor infiltration in vivo.
Methods: Bone marrow cells from healthy wildtype Balb/c
mice were differentiated into macrophages and subsequently polarized to the M1 or M2 phenotype in the presence of
different FAK inhibitor concentrations (VS-4718, small molecule
FAK/PYK2 inhibitor). The expression of signature M1 and M2
markers was measured after culture using flow cytometry. In
addition, SCID mice bearing mesothelioma xenografts were
treated with VS-4718 (50 mg/kg, po) twice daily. Mice were sacrificed after 10 days, tumors were harvested, and macrophage
abundance was assessed by immunofluorescence staining of
the F4/80 marker.
Results: Addition of VS-4718 to the standard M1 macrophage
culture resulted in a more prominent M1 polarized phenotype, demonstrated by a higher expression of MHCII and
PD-L1 compared to the M1 macrophage control. Furthermore,
expression of the M2 markers CD206 and CD115 was downregulated on macrophages in the presence of VS-4718 compared
to the M2 macrophage control. In vivo, there was a significant
downregulation of macrophages in the tumors of mice treated
with VS-4718. Growth of the MM87 xenograft tumors was also
significantly inhibited by VS-4718 treatment.
Conclusion: The effect of FAK inhibition on macrophages was
investigated both in vitro and in vivo in mesothelioma mouse
models. We demonstrate that by inhibiting FAK signaling, the
macrophage phenotype can be skewed towards a more M1like pro-inflammatory phenotype in vitro. Furthermore, in a
mesothelioma mouse model, total TAM numbers were diminished after treatment with the FAK inhibitor. Further studies are
needed to confirm these preliminary results and to investigate
how FAK inhibitors can be optimally used to target TAMs in the
mesothelioma microenvironment. The current study illustrates
the potential to use FAK inhibitors in novel ways in the clinic to
enhance anti-tumor immunity, for example, in combination with
checkpoint inhibitors.
however local immunosuppressive mechanisms could hamper
their efficacy. Macrophages are abundantly present within the
mesothelioma microenvironment. This study investigates the
influence of the macrophage phenotype and their capacity to
inhibit local immune responses and the decisive role of pleural
effusion (PE) in this regard.
Methods: Healthy monocytes derived from a buffy coat were
used to culture macrophages in the presence of PEs (n=6) to
create a mesothelioma environment. Macrophage phenotype
was investigated using RT-PCR. Macrophages and healthy T
cells were co-cultured in the presence of PEs and accompanying mesothelioma cell line supernatants (n=6). T cell proliferation after co-culture was calculated as output measure using
flow cytometry. The levels of 11 pro- and anti-inflammatory
cytokines and the prostanoid prostaglandin E2 (PGE2) were
measured in PEs (n=6) and accompanying tumor cell line supernatants (n=6) using a magnetic-bead based multiplex assay
and ELISA. The presence and phenotype of macrophages and T
cell subsets was measured in the PE of mesothelioma patients
(n=30) using flow cytometry.
Results: PE induced a tumor promoting M2 phenotype in
macrophages. Macrophages cultured in the presence of PEs
firmly suppressed T cell proliferation during co-culture (p<0.05,
compared to co-culture in the presence of normal human
serum). The mesothelioma cell line supernatants did not exert
this effect. The level of PGE2 present in PEs correlated with the
induction of the suppressive capacity of macrophages, this correlation could not be found with any of the measured cytokines.
Macrophages isolated from PEs of mesothelioma patients
displayed an M2 phenotype and were negatively correlated with
T cells in vivo (rho -0.90, p<0.001).
Conclusion: The current study demonstrates that macrophages in pleural effusion can play a pivotal role in directly
hampering the anti-tumor T cell immune response. This emphasizes the potential of macrophages as a therapeutic target in
mesothelioma and indicates that the presence and phenotype
of macrophages in pleural effusion should be taken into consideration in the application of (intrapleural) immunotherapies,
besides being also a potential prognostic measure.
Keywords: tumor-associated macrophages, focal adhesion
kinase, mesothelioma mouse model
Keywords: tumor-associated macrophages, tumor microenvironment, Pleural effusion
PP02.22: PLEURAL EFFUSION OF PATIENTS
WITH MALIGNANT MESOTHELIOMA INDUCES
MACROPHAGE-MEDIATED T CELL SUPPRESSION
PP02.23: THE ABSCOPAL EFFECT OF
RADIOTHERAPY IN THE CONTEXT OF CHECKPOINT
BLOCKADE IN A MOUSE MESOTHELIOMA MODEL
Lysanne Lievense, Koen Bezemer, Robin Cornelissen,
Margaretha Kaijen-Lambers, Joachim Aerts, Joost Hegmans
Alistair M. Cook1, Jason Waithman2, Willem J. Lesterhuis1,
Martin Ebert3 , Roslyn Francis1, Scott Fisher1, Alison Mcdonnell1,
Sean Bydder4 , Sally M. Lansley5, Bruce Robinson1, Richard
Lake1, Anna Nowak1
NETHERLANDS
School of Medicine and Pharmacology, The University of Western
Australia, Crawley, WA, AUSTRALIA, 2Telethon Kids Institute,
Subiaco, WA, AUSTRALIA, 3School of Physics, The University of
Western Australia, Crawley, WA, AUSTRALIA, 4School of Surgery,
The University of Western Australia, Crawley, WA, AUSTRA1
Objectives: Clinical studies have demonstrated beneficial
effects of immunotherapy in malignant pleural mesothelioma. The pleural cavity, close to the target tumor, seems an
attractive compartment to administer these type of therapies,
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LIA, 5Pleural Medicine Unit, Institute for Respiratory Health,
Perth, WA, AUSTRALIA
Objectives: In people with advanced cancer, including
mesothelioma, local radiotherapy is often used to alleviate
discomfort from specific tumour sites. Very rarely, radiotherapy
to one site can induce shrinkage of tumours elsewhere in the
body. Termed the ‘abscopal’ effect, this has been attributed to
the way that radiotherapy modifies the tumour microenvironment, and in particular the manner in which it kills cancer cells.
This ‘immunogenic’ cell death can release damage-associate
molecular patterns (DAMPs) that can activate dendritic cells
and in turn tumour-specific cytotoxic T cells, thereby generating
an immune response by turning a tumour into its own ‘vaccine’.
However, the fact that the abscopal effect is so rare is likely due
to localised or systemic immune suppression. Crucially, the abscopal effect is more frequently observed in patients receiving
immunotherapy, particularly during recent trials of immunological checkpoint antibodies e.g. CTLA-4 or PD-1 pathway blockade, and there is emerging evidence that local radiotherapy can
enhance the efficacy of immunotherapy. However, immunological synergy between radiotherapy and checkpoint blockade
remains poorly studied, particularly in thoracic cancers. Here,
we present a strategy to study the abscopal effect by combining radiotherapy and immune checkpoint blockade in a mouse
model of mesothelioma.
Results: Preliminary data will be presented.
Conclusion: We will develop a relevant, tractable preclinical
model of mesothelioma in which to study the interaction of
radiotherapy and the immune system. The intrapleural/flank
tumour model in particular is highly relevant to the clinical
setting.
Keywords: radiotherapy, intrapleural, Preclinical, Immunotherapy
Methods: This project uses CT-targeted precision irradiation,
as the best available pre-clinical equivalent to clinical radiotherapy, to irradiate single tumours in mice bearing dual mesothelioma tumours on opposite flanks (Figure 1). We will use flow
cytometry to characterise both local and systemic immunological changes occurring in response to radiotherapy, including
the expression of the checkpoint molecules CTLA4, PD-1/
PD-L1, OX40 and TIM3. This will be followed by an evaluation of
which checkpoint antibodies are most effective when combined
with targeted radiotherapy, initially as individual treatments and
proceeding to double combinations in addition to radiotherapy.
The most effective combinations will be tested by irradiating
single flank tumours in mice bearing intrapleural tumours,
mimicking pleural disease with irradiated chest wall metastasis.
PET/CT and MRI will be used to assess tumour growth in mice
with intrapleural mesothelioma. iMig2016.ORG
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ABSTRACT BOOK
PP02.26: TARGETED DEPLETION OF REGULATORY
T CELLS IN MESOTHELIOMA
Scott Fisher, Jessica Solin, Wayne J. Aston, Amanda Cleaver,
Willem J. Lesterhuis, Andrea Khong, Bruce Robinson, Richard
Lake
Perth, WA, AUSTRALIA
Objectives: Recent advances in the use of checkpoint blockade immunotherapies such as CTLA-4 and PD-1 that act to
block different aspects of negative T cell regulation are beginning to be explored in mesothelioma. Regulatory T cells (Treg)
play an important role in suppressing anti-tumour immunity
and their depletion has been linked to improved outcomes. We
used the BALB/c FoxP3.dtr transgenic mice to specifically target
and deplete Treg via Diphtheria toxin (DTX) to better understand
the role of Treg in limiting the efficacy of different mesothelioma
treatment protocols.
Methods: Mice bearing AB1 mesotheliomas were subject to
different treatment protocols, with or without DTX Treg depletion and tumour growth and survival monitored. Polychromatic
flow cytometry analysis was used to confirm Treg depletion,
phenotype immune cell populations and assess immune correlates with the outcome of each treatment protocol
Results: DTX specifically depletes Treg in FoxP3.dtr mice bearing mesothelioma tumours. Treg depletion lasts around three
days and occurs in a dose dependent manner with maximum
depletion observed one day after treatment. Treg depletion
did not alter the proportion of non-Treg CD4 or CD8 effector
T cells, but Treg recovery was associated with a significant
increase in effector T cell activation and proliferation. Systemic
administration of DTX caused depletion of Treg in all tissues,
although the kinetics of Treg recovery in the tumour was slower
than other organs and the level of CD8 T cell activation was less
pronounced.
PP02.27: LEGAL CLAIMS FOR ASBESTOSIS IN THE
NETHERLANDS POSSIBLE SINCE 2014: HOW IT
WORKS
Wanda Hagmolen Of Ten Have1, Jos Rooijackers2, Wieneke
Buikhuisen3 , Jan Grutters4 , Sjaak Burgers3
Pulmonology, Radboudumc, Nijmegen, NETHERLANDS, 2Institute For Risk Assessment Sciences, UMCU, Utrecht, NETHERLANDS, 3Thoracic Oncology, Netherlands Cancer Institute, Amsterdam, NETHERLANDS, 4Pulmonology, St. Antonius hospital,
Nieuwegein, NETHERLANDS
1
Objectives: Patients with mesothelioma can claim financial
compensation via the Dutch institute for asbestos victims.
In March 2014 the Dutch Minister of Social Affairs and Employment announced that the same should be possible for
patients with asbestosis. The diagnosis of asbestosis can only
made definitive with histological (biopsy) material. However, in
contrary to patients with mesothelioma histological material
is almost never available. The risks of invasive biopsies do not
weigh against the benefits because there is no specific treatment available to fight asbestosis. The so-called ‘Mesothelioma
Clinical Expert Panel of the Dutch Thoracic Society’ was asked
to assess the available clinical data of the victims to assess
whether the victim has (putative) asbestosis or not.
Methods: A flow diagram of the diagnostic work-up was
constructed for patients that applied to the Dutch institute for
asbestos victims (see figure).
(See next page.)
Conclusion: This pre‑clinical study suggests that mesothelioma is likely to respond to Treg depletion and therefore targeted
removal of Treg could be an effective therapeutic strategy. A
major problem in translating this into clinical reality is identifying markers that specifically target Treg without affecting effector T cells. In our related studies (abstract 198), we have shown
that tumour resident Treg express high levels of the immune
checkpoint molecules CTLA-4, GITR and OX40, while tumour
infiltrating lymphocytes express PD-1 and TIM-3. Taken together, these data suggest that specific combinations of checkpoint
blockade immunotherapies could be designed to specifically
target Treg and simultaneously enhance effector T cell function
to generate effective therapy for mesothelioma.
Keywords: Immunotherapy, Treg, mesothelioma
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ABSTRACT BOOK
Results: Over 180 claims have been received since April 1th
2014. Until now about 40 victims might have asbestosis and
received € 19.417 in 2015. The majority of asbestos victims who
applied for the financial compensation had pleural plaques and
no fibrosis.
Conclusion: In 2014 the Dutch government extended existing
regulation for financial compensation for asbestos victims to
patients with asbestosis. Until now about 40 victims succeeded
in their claim. Updated data will be presented at the iMig 2016.
PP02.28: EVALUATION OF CURCUMIN I.P. FOR THE
TREATMENT OF SARCOMATOID MESOTHELIOMA,
EXPERIMENTAL STUDY ON A RAT MODEL
Daniel L. Pouliquen1, Béatrice Nawrocki-Raby2, Joëlle Nader1,
Myriam Robard3 , Philippe Birembaut2, Marc Grégoire1
UMR 892 INSERM / 6299 CNRS, Nantes, FRANCE, 2UMRS-903
INSERM, Reims, FRANCE, 3Plateforme MicroPICell, SFR F. Bonamy, Nantes, FRANCE
1
Objectives: Both the literature and clinical trials have confirmed the potential of curcumin against various types of
cancers. Additionally, the epigenetic modulation of target genes
by this molecule has been recently highlighted, pointing out the
interest of drugs relevant to polypharmacology. In this study, a
rat model of sarcomatoid mesothelioma, mimicking some of the
worse clinical conditions faced in clinics, was used to evaluate
the therapeutic potential of curcumin administered intraperitoneally.
Methods: The M5-T1 cell line, selected from a collection
established from F344 rats induced with crocidolite given i.p.
over a period of 136 to 415 days, was inoculated intraperitoneally in syngeneic rats, producing in three weeks macroscopic
tumors growing on the omentum together with metastases in
several normal tissues including the liver, diaphragm, pancreas
and spleen. The potential of i.p. administration of curcumin to
kill tumor cells in vivo and induce infiltration with immune cells,
was evaluated in comparison with a reference epigenetic drug,
SAHA, in tumor-bearing rats.
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ABSTRACT BOOK
Results: Both agents administered at days 21 and 26 after
tumor challenge produced necrosis within the solid tumors at
day 28. Quantification of the surfaces covered by necrotic cells
demonstrated that the extent of necrosis within the invaded
omentum was significantly higher after treatment with curcumin compared with treatment with SAHA. Quantification of
residual tumor cells within these necrotic areas also revealed
their number was very significantly lower after treatment with
curcumin relative to SAHA. After treatment with curcumin, the
tumor tissue at the periphery of necrotic areas was characterized by the presence of numerous mononuclear phagocytic
cells. In contrast, treatment with SAHA was characterized by
the presence of foci of resistant cells within the tumor tissue
and infiltration by numerous isolated CD8+ cells. The treatment
of tumor bearing rats one week after tumor challenge with
four successive injections of 1.5 mg/kg curcumin on days 7, 9,
11 and 14 dramatically reduced the total tumor mass at day
16. Clusters of CD8+ lymphocytes were also observed at the
periphery of small tumor masses remaining in the peritoneal
cavity where the number of mitosis per field was significantly
reduced compared with tumors in untreated rats.
Conclusion: These data open up interesting new prospects
for the therapy of malignant mesothelioma with curcumin or its
many derivatives administered intracavitary.
Keywords: Peritoneal mesothelioma, Sarcomatoid, Curcumin,
Rat tumor model
PP02.29: BIPHASIC PLEURAL MESOTHELIOMA:
UNUSUAL CLINICAL BEHAVIOR
Abdulhadi Almutairi, Ahsan Cheema, Ikram Chaudhry
Surgery, King Fahad Specialist Hospital, Dammam, SAUDI
ARABIA
Objectives: Biphasic pleural mesothelioma is a fatal disease,
which shows aggressive clinical behavior with low survival
potential. We report two cases treated only with external beam
radiation and both patients survived for 4 and 5 years, respectively.
Methods: Retrospective chart review of two cases that presented with symptomatic unilateral pleural effusion and pleural
nodularity. Both patient underwent uniportal VATs pleural biopsy and both biopsies confirmed the diagnosis of biphasic pleural
mesothelioma. Both patients were offered surgical resection
but refused and opted for external beam radiation Therapy
(EBRT). In both patients, no asbestos fibers were identified on
pathology slides review.
Results: Case 1: A 65-year-old heavy smoker male with no asbestos exposure risk presented in 2010 with shortness of breath
and a right-sided pleural effusion (Fig 1a). Patient was treated
with EBRT with a total dose of 24 Gy over 2 weeks. The patient
remained asymptomatic for four years. Periodic imaging, both
chest x-ray and CT scan, confirmed no effusion recurrence (Fig
1b). Patient presented in 2015 with brain metastasis and passed
away few days after presentation. Case 2: A 48-year-old non-
smoker female patient with no exposure risk, presented in 2011
with chest discomfort and a left sided pleural effusion (Fig 2a).
She was offered surgical resection but refused and treated with
EBRT in similar manner. Patient still alive and symptom-free till
the time of this abstract (Fig 2b).
Conclusion: Radiation therapy may have a significant role that
prolongs survival in subset of patient with biphasic pleural mesothelioma. Further studies are needed to explore prognostic
risk factors and stratify different spectrum of biphasic pleural
mesothelioma.
Keywords: mesothelioma, Survival, biphasic, radaition
PP02.30: MARS 2: A FEASIBILITY STUDY
COMPARING (EXTENDED) PLEURECTOMY
DECORTICATION VERSUS NO PLEURECTOMY
DECORTICATION
Eric Lim
Royal Brompton Hospital, London, UNITED KINGDOM
Objectives: The aim of the MARS 2 study is to determine if it is
feasible to recruit patients with malignant pleural mesothelioma
with disease amenable to surgical resection into a randomised
trial of (extended) pleurectomy decortication (lung sparing
surgery) versus no surgery. The feasibility component will also
assess if there is any evidence of harm associated with (extended) pleurectomy decortication.
Methods: Patients with a histological confirmation of mesothelioma with disease confined to one hemi-thorax are eligible
for enrolment. Patients are ineligible if they are unable to give
informed consent or are unwilling to be randomised or if they
have disease that is not deemed to be surgically resectable, an
ECOG status 2 or more, a predicted pre-operative FEV1 or TLco
less than 20%, severe heart failure, end stage kidney failure,
liver failure or are already participating in another interventional
clinical trial. All patients will receive the usual standard of care
chemotherapy. After 2 cycles, participants will be re-assessed
by CT to screen for progressive disease. Patients with no
evidence of disease progression beyond the limits of surgical
resection will be randomised to either: A). (Extended) pleurectomy decortication or B). No surgery. All patients will then receive
the remaining 4 cycles of chemotherapy.
Results: Two lead surgical centres in the UK, Leicester and
Sheffield, are performing the surgery for this feasibility phase.
10 other medical centres in the UK are also currently recruiting
patients with another 10 medical centres in set-up. To date, 19
patients have been enrolled and 7 patients randomised.
Conclusion: The results from the MARS 2 feasibility study
will determine if it is possible to recruit patients with malignant pleural mesothelioma with disease amenable to surgical
resection into a randomised trial of (extended) pleurectomy
decortication (lung sparing surgery) versus no surgery. The feasibility study will also assess if there is any evidence of harm. If
feasibility and safety are demonstrated, the MARS 2 feasibility
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ABSTRACT BOOK
study will be extended to a larger study powered to assess
survival and quality of life.
Keywords: Extended Pleurectomy Decortication, Lung sparing
surgery
PP02.31: MESOTRAP: A FEASIBILITY STUDY
OF INDWELLING PLEURAL CATHETER VERSUS
VAT PLEURECTOMY FOR TRAPPED LUNG IN
MESOTHELIOMA
Robert C. Rintoul1, Angela Tod2, Pasupathy Sivasothy3 , Graham
Sherlock-Brown4 , David Waller5, Aman Coonar1, John Edwards6 ,
Nick Maskell7, Naj Rahman8 , Julia Fox-Rushy9, Linda Sharples10
Thoracic Oncology, Papworth Hospital NHS Foundation Trust,
Cambridge, UNITED KINGDOM, 2University of Manchester, Manchester, UNITED KINGDOM, 3Cambridge University Hospitals,
Cambridge, UNITED KINGDOM, 4Patient Public, Glenfield Hospital, Leicester, UNITED KINGDOM,5Glenfield Hospital, Leicester, UNITED KINGDOM, 6Northern General Hospital, Sheffield,
UNITED KINGDOM, 7University of Bristol, Bristol, UNITED KINGDOM, 8University of Oxford, Oxford, UNITED KINGDOM, 9Brunel
University, London, UNITED KINGDOM, 10University of Leeds,
Leeds, UNITED KINGDOM
1
Objectives: Pleural effusion in malignant pleural mesothelioma (MPM) is often managed by promoting a talc pleurodesis
between the visceral and parietal pleura. However if the pleura
are unable to appose, an effective pleurodesis is unlikely. In
MPM tumour is commonly present over the visceral surface of
the lung which prevents the lung from re-inflating - so called
‘trapped lung’ (TL). In this situation pleural fluid usually recurs
and patients go through repeated cycles of fluid drainage and
re-accumulation. TL is a challenging management issue causing
significant morbidity and in case series of TL, 13-37% have
been due to MPM. Some authorities recommend placement
of an indwelling pleural catheter (IPC) under local anaesthesia
to facilitate repeated drainage in the community. Others have
advocated general anaesthesia video-assisted thoracoscopic
partial pleurectomy/decortication (VAT-PD) to remove as much
tumour from the visceral pleural surface as possible to allow
the lung to re-expand to permit a pleurodesis. Recognising that
management of dyspnoea due to TL lung in MPM is a significant
unmet need, our ultimate aim is to undertake a randomised
controlled trial comparing VAT-PD with IPC. However, prior to
undertaking a full phase III study we need to address several
uncertainties: i) How prevalent is TL in MPM? ii) Will patients
accept randomisation to IPC or VAT-PD? iii) What is the standard deviation of Visual Analogue Scale scores for dyspnoea
and chestpain in each treatment group? (This will be used to
estimate parameters that will be included in the sample size
estimates for a full trial). Therefore initially we will undertake
a feasibility study, the primary objective of which will be to
determine the ability to randomise patients into a trial of VATPD versus IPC in patients with TL and pleural effusion due to
MPM. To help inform the design of a phase III trial secondary
objectives will be: i) The prevalence of TL in patients with MPM
ii) Visual Analogue Scale scores for dyspnoea and chest pain
and the patterns of change over time in each treatment group
iii) Quality of Life data at baseline, 1, 3, 6 and 12 months post
randomisation iv) Collection and documentation of Adverse
Events v) Health economic analysis
Methods: Six UK thoracic surgical centres with expertise in
performing both IPC and VAT-PD along with linked non-surgical
referring hospitals, all of whom are previous/existing mesothelioma study collaborators, will randomise 36 patients (1:1)
in 18 months. Inclusion criteria: confirmed MPM; TL (>20% of
one hemithorax without lung markings on chest x-ray) following fluid drainage; deemed suitable for VAT-PD by a thoracic
surgeon. Following ethics approval, a trial management group,
trial steering and data monitoring committees will provide
governance. The study will be registered with an International
Standard Randomised Controlled Trial Number (ISRCTN) and
ClinicalTrials.gov.
Results: Section not applicable
Conclusion: The feasibility study will provide information as
to a) whether a full randomised controlled trial is achievable in
a reasonable time frame, b) the number of patients required.
A full trial will determine best management of trapped lung in
MPM.
Keywords: Malignant pleural mesothelioma, Indwelling Pleural
Catheter, Trapped Lung, Video Assisted Thoracoscopic Pleurectomy
PP02.32: LUME-MESO: A PLACEBO-CONTROLLED
PHASE II/III STUDY OF NINTEDANIB +
PEMETREXED/CISPLATIN FOLLOWED BY
MAINTENANCE NINTEDANIB
Giorgio V. Scagliotti1, Rabab M. Gaafar2, Anna Nowak3 , Martin
Reck4 , Jan Van Meerbeeck5, Arsene-Bienvenu Loembe6 , Ute
Von Wangenheim7, Derek Velema8 , Sanjay Popat9
Department of Oncology, University of Turin, Torino, ITALY, 2National Cancer Institute, Cairo University, Cairo, EGYPT, 3School of
Medicine and Pharmacology Qeii, Medical Centre Unit, University
of Western Australia, Crawley, WA, AUSTRALIA, 4Department of
Thoracic Oncology, Lung Clinic Grosshansdorf, Member of the
German Center for Lung Research (DZL), Grosshansdorf, GERMANY, 5Department of Thoracic Oncology, University Hospital Antwerp, Edegem, BELGIUM, 6Boehringer Ingelheim B.V., Alkmaar,
NETHERLANDS, 7Boehringer Ingelheim Pharma GmbH & Co. KG,
Biberach, GERMANY, 8Boehringer Ingelheim (Canada) Ltd./Ltée,
Burlington, ON, CANADA, 9Royal Marsden Hospital NHS Foundation Trust, London and Surrey, UNITED KINGDOM
1
Objectives: Pemetrexed/cisplatin doublet is considered the
front-line standard-of-care treatment for patients with unresectable malignant pleural mesothelioma (MPM) and yields a median overall survival time of roughly one year. Additional improvements in therapy are clearly needed. Nintedanib is an oral,
twice-daily, angiokinase inhibitor targeting vascular endothelial
growth factor (VEGF) receptors 1–3, platelet-derived growth
factor receptors α/β, and fibroblast growth factor receptors
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ABSTRACT BOOK
1–3, as well as Src and Abl kinase signaling, which are involved
in regulating tumour angiogenesis, growth, and metastasis of
MPM. Inhibition of the VEGF pathway as a successful treatment
approach for MPM has been previously demonstrated with
bevacizumab in the Phase III IFCT-GFPC-0701 MAPS trial (J Clin
Oncol 33, 2015: suppl; abstr 7500). Nintedanib has shown clinical benefit and a manageable safety profile in several tumour
types, and can be co-administered with various anticancer
drugs. Nintedanib (VARGATEF®) in combination with docetaxel
is approved in the European Union and additional countries
for the treatment of patients with locally advanced, metastatic
or locally recurrent NSCLC of adenocarcinoma histology after
first-line chemotherapy. Nintedanib (Ofev®) monotherapy is
approved in the USA and EU for idiopathic pulmonary fibrosis.
LUME-Meso is an international, double-blind, randomised, multicentre, placebo-controlled Phase II/III study to evaluate the
efficacy and safety of nintedanib combined with pemetrexed/
cisplatin for the treatment of unresectable MPM. Following a
data review by the Data Monitoring Committee (DMC) after
all planned Phase II patients had been enrolled, the Phase II
exploratory study was changed to a confirmatory Phase III trial.
The trial is ongoing, remains blinded and the primary analysis
has not been conducted.
Methods: Chemo-naïve patients from 25 countries worldwide
(≥18 years of age, ECOG PS 0–1, and histologically confirmed
epithelioid or biphasic MPM; 87 patients in Phase II and 310 to
450 patients in Phase III) will be randomised in a 1:1 ratio to
receive up to six cycles of pemetrexed (500 mg/m2)/cisplatin
(75 mg/m2) on Day 1 administered along with nintedanib (200
mg bid) or placebo from Days 2–21. Patients without disease
progression (PD) will continue to receive maintenance treatment with either nintedanib or placebo until PD. The primary
endpoint is progression-free survival (PFS), with overall survival
(OS) as the key secondary endpoint. The study will use an
adaptive design strategy, with sample size reassessment by
an external DMC during the trial based on an interim analysis,
to ensure sufficient power for PFS and OS. Depending on the
treatment effect, a maximum of 450 additional patients will be
randomised. Additional secondary endpoints include objective
tumour response and disease control according to modified
RECIST. Other assessments include frequency and severity
of adverse events and changes in laboratory parameters to
measure safety, baseline change in forced vital capacity as a
measure of pulmonary function, health-related quality of life
and exploratory biomarker analyses that will focus on exploring
predictive biomarkers in tumour and blood specimens. The
study is currently enrolling patients into Phase III. Clinical trial
identifier: NCT01907100.
Keywords: Phase II/III clinical trial, Nintedanib, unresectable
malignant pleural mesothelioma, angiogenesis
PP02.33: AUTOLOGOUS DENDRITIC CELL
IMMUNOTHERAPY LOADED WITH AN ALLOGENIC
TUMOR CELL LYSATE IN PATIENTS WITH
MESOTHELIOMA
Joachim Aerts1, Paul Baas2, Rosanna Berardi3 , Dean Fennell4 ,
Jan Van Meerbeeck5, Arnaud Scherpereel6 , Joost Hegmans1
NETHERLANDS, 2Thoracic Oncology, Netherlands Cancer Institute, Amsterdam, NETHERLANDS, 3University hospital Ancona,
Ancona, ITALY, 4Cancer Studies, University of Leicester, Leicester, UNITED KINGDOM, 5Thoracic Oncology/MOCA, Antwerp
University Hospital, Antwerp, BELGIUM, 6Pulmonary and Thoracic
Oncology Department, Hospital of the University (CHU) of Lille,
Lille cedex, FRANCE
1
Objectives: Immune therapy with checkpoint inhibition has
shown clinical efficacy in a number of malignancies including
mesothelioma. However despite impressive clinical responses, a substantial proportion of patients do not respond to this
treatment. Althoug this number of responding patients may be
increased by combining different checkpoint inhibitors it has
now become clear that in a substantial proportion of patients
cell based therapy is neccessary to yield an effective immune
response. We have shown in former trials that dendritic cell
immunotherapy is an effective cell therapy to induce an anti-tumor T-cell response in patients. We have now been granted
by the Europian Union, a research proposal to investigate the
effect of dendritic cell based immunotherapy compared to BSC.
The co-primary objectives are to determine the effect on overal
survival and the percentage of patients without progression at
12 months in patients with mesothelioma who were not progressing after platinum pemetrexed chemotherapy
Methods: Patients, non progressing after 4-6 cycles platinum
pemetrexed chemotherapy, can be included in the trial. Patients
will be randomised 1:1 to either dendritc cell therapy or best
supportive care (BSC). In case of active treatment, after the
completion of chemotherapy, patients will undergo a leucopheresis, during which monocytes are isolated. From these monocytes immature dendritic celles are generated. These immature
dendritic cells willl be loaded with an allogenic lysate generated
from mesothelioma patients derived clinical grade cellines
(Pheralys). We plan to include around 200 patients to achieve
a hazard ratio below 0.7 for the overal survial comparing active
treatment and BSC.
Results: No results are present
Conclusion: This study will invetigate the effect of dendritic
cell therapy compared to BSC. The planned start of the study is
in 2016 and accrual is planned untill 2018.
Keywords: Immunotherapy, dendritic cell
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ABSTRACT BOOK
PP02.34: RANDOMISED PHASE II TRIAL
OF VINORELBINE AS 2ND-LINE THERAPY
FOR PATIENTS WITH MALIGNANT PLEURAL
MESOTHELIOMA – VIM TRIAL
ed by the Wales Cancer Trials Unit, Cardiff, UK. This abstract is
submitted on behalf of the VIM TMG.
Keywords: Malignant pleural mesothelioma (MPM), Vinorelbine
Lisette S. Nixon1, Georgina Gardner1, Angela Casbard1, Jason F.
Lester2, Dean Fennell3
Wales Cancer Trials Unit, Centre For Trials Research, Cardiff
University, Cardiff, UNITED KINGDOM, 2Velindre Cancer Centre,
Velindre NHS Trust, Cardiff, UNITED KINGDOM, 3Cancer Studies,
University of Leicester, Leicester, UNITED KINGDOM
1
Objectives: Malignant pleural Mesothelioma (MPM) is increasing worldwide. However there is no approved therapy in the
second-line setting. Vinorelbine exhibits promising activity in a
proportion of patients, although there has been no randomised
evaluation or validation of biomarkers to support patient stratification. We have reported that BRCA1 is an essential regulator of
MPM sensitivity to vinorelbine, and its expression is lost in approximately 38%. The aim of the VIM clinical trial is to establish
the anti-tumour activity of vinorelbine as measured by overall
survival (OS), time from randomisation to death. Secondary outcome measures include progression-free survival and objective
response rate using modified RECIST, safety, tolerability and
feasibility. Tumour and blood samples will be collected for
future translational research including investigation of BRCA1
expression as a putative predictor of vinorelbine sensitivity.
Methods: A UK multicentre open-label randomised phase
II trial. Eligible patients will have histological diagnosis of
mesothelioma, received at least one line of platinum doublet
based chemotherapy as standard, be 18 years or older, have
measurable lesions by modified RECIST, radiological evidence
of disease progression and given informed consent. Patients
will be randomised to either control (ASC) or ASC plus vinorelbine using a 1:2 allocation ratio. ASC will be administered as
per local practice, continuing follow-up for at least 18 months.
Vinorelbine will be administered at a dose of 60mg/m2 po od on
day 1 (equivalent to 25mg/m2 iv day 1) weekly for the first cycle
(3 weeks), then subsequently increased to 80mg/m2 weekly
(equivalent to 30mg/m2 iv), in the absence of haematological
toxicity for subsequent cycles. Patients will continue chemotherapy until evidence of radiological progression, unacceptable
toxicity, or patient withdrawal. The primary endpoint of the trial
is overall survival, with secondary endpoints of tolerability,
response rate, change in tumour volume and progression-free
survival. A subgroup analysis will explore the effect of BRCA1
expression on overall survival. The median OS for patients in
the control arm is expected to be 9.7 months. With 90% power
and a one-sided α of 0.2, 104 events and 133 patients are
required to detect a hazard ratio of 0.65, based on the logrank
test. The sample size was inflated to allow sub-group analysis of
BRCA1 expression at the end of the trial: 133 will be recruited to
the vinorelbine arm and 67 to the control arm.
Results: Not applicable.
Conclusion: Not applicable. The UK National Cancer Research
Institute Lung Clinical Studies Group have helped to develop
the VIM clinical trial. The study is funded by a research grant
from Cancer Research UK (CRUK/12/056) and vinorelbine is
supplied and distributed free of charge from Pierre Fabre Ltd.
The trial is sponsored by University of Leicester, and coordinat-
PP02.35: A RANDOMIZED STUDY OF AMATUXIMAB
WITH PEMETREXED AND CISPLATIN AS FRONTLINE THERAPY FOR SUBJECTS WITH PLEURAL
MESOTHELIOMA
Bruce A. Wallin1, Kimberly Hoffman1, Megan Mclaughlin1, Raffit
Hassan2
Clinical Development, Morphotek, Inc, Exton, PA, UNITED
STATES OF AMERICA, 2Thoracic and Gastrointestinal Oncology Branch, National Cancer Institute, Bethesda, MD, UNITED
STATES OF AMERICA
1
Objectives: Amatuximab is a chimeric monoclonal antibody
that binds to mesothelin, which is highly expressed in malignant
mesothelioma. Amatuximab was studied in a Phase 2 malignant
pleural mesothelioma (MPM) trial which demonstrated that the
safety profile of amatuximab in combination with P/C was consistent with that seen previously for the P/C regimen. Although
PFS was not significantly different from historical results of P/C
alone, the median OS was 14.8 months (as compared to 13.3
months for P/C). The post-hoc PK/PD analysis demonstrated
that amatuximab trough concentrations were a significant
predictor of both OS and PFS with higher amatuximab concentrations associated with longer OS (583 days; p=0.0202) and
PFS (238 days; p<0.001). The primary objective of the study
is to demonstrate whether weekly amatuximab, 5 mg/kg, in
combination with pemetrexed and cisplatin, has superior OS
compared with pemetrexed and cisplatin and placebo in subjects with unresectable MPM.
Methods: This Phase 2, double-blind, placebo-controlled study
(NCT02357147) is ongoing in 6 countries. Eligible patients (pts)
have a confirmed diagnosis of unresectable, epithelioid MPM
with measurable disease at screening, and an ECOG Performance Status 0 or 1. Prior chemotherapy, radiation, or surgery
with curative intent is not allowed. 560 subjects will be randomized 1:1 to receive weekly amatuximab, 5 mg/kg or saline placebo IV in combination with pemetrexed, 500 mg/m2, and cisplatin, 75 mg/m2, on Day 1 of each 21-day cycle for 6 cycles. Pts
with tumor response or stable disease continue amatuximab
monotherapy or placebo weekly until disease progression or
study termination. The primary endpoint will be a comparison
of overall survival between amatuximab and placebo groups
in the intent-to-treat population using the unstratified log-rank
test. Secondary endpoints include progression-free survival,
overall response rate, and duration of response. Safety data will
include the usual metrics, including human anti-drug antibody
measurements. An interim analysis for futility will be performed
by an Independent Assessment Group after approximately 86
OS events are accrued. An IDMC will review the results of the
interim analysis for futility and provide final recommendations.
Six of the planned 560 pts are enrolled.
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ABSTRACT BOOK
Keywords: mesothelioma, amatuximab
PP02.36: SWITCH-MAINTENANCE WITH
GEMCITABINE FOR PATIENTS WITH MALIGNANT
MESOTHELIOMA: FEASIBILITY OF A RANDOMIZED
PHASE II STUDY
Josine Quispel-Janssen1, Paul Baas1, Wieneke Buikhuisen1,
Vincent Vd Noort2, Joachim Aerts3 , Robin Cornelissen3 , Harry
Groen4 , Bonne Biesma5, Robbert Van Heemst6 , Magdolen El
Soud Youssef7, Jeske Staal-Vd Brekel8 , Sjaak Burgers1
Thoracic Oncology, Netherlands Cancer Institute, Amsterdam,
NETHERLANDS, 2Statistics, Netherlands Cancer Institute, Amsterdam, NETHERLANDS, 3Dept. Of Pulmonary Medicine, Erasmus
Medical Center, Rotterdam, NETHERLANDS, 4Dept Of Pulmonary
Medicine, University Medical Center Groningen, Groningen,
NETHERLANDS, 5Dept Of Pulmonary Medicine, Jeroen Bosch
Hospital, Den Bosch, NETHERLANDS, 6Dept. Of Pulmonary
Medicine, Deventer Hospital, Deventer, NETHERLANDS, 7Dept Of
Pulmonary Medicine, Maxima Medical Center, Veldhoven, NETHERLANDS, 8Dept. Of Pulmonary Medicine, Zorggroep Twente,
Almelo, NETHERLANDS
1
Objectives: The prognosis of malignant mesothelioma (MM)
remains poor in spite of an increasing use of second line
treatment. No randomized clinical trial in second line has
demonstrated improved survival yet. There is scarce data on
maintenance chemotherapy in MM showing that maintenance
with pemetrexed after 6 cycles combination therapy is feasible. However, studies from other solid tumors suggest that
switch-maintenance with a non-cross resistant drug may be
more effective. Gemcitabine has extensively been tested in
MM in combination with cis- or carboplatin and has resulted in
response rates ranging from 12% to 48%. Here, we describe
the trial design and first toxicity results of a switch-maintenance
trial with gemcitabine monotherapy.
Methods: In this Dutch multi-center, randomized phase II
study, patients without progression after first line chemotherapy, are randomized to receive either gemcitabine 1250mg/
m2 i.v. on day 1+8 every three weeks plus best supportive care
(BSC), or BSC alone. Primary endpoint is progression free survival (PFS). Secondary objectives include response rate, overall
survival, toxicity and identification of potential biomarkers for
response. A total of 124 patients will be recruited to achieve a
power of 90% with a false-positive error of 0.1.
Results: Since the start of the study in March 2014 35 patients
have been randomized. Seventeen received at least one cycle
of gemcitabine. A total of 34 adverse events were seen in 11
patients; 6 receiving gemcitabine and 5 BSC. In the gemcitabine
arm , two patients had an infection leading to hospitalization.
One of these patients died due to sepsis. The other recovered
without further symptoms. Besides this, toxicity so far is mild
and manageable.
Keywords: maintenance, randomized, gemcitabine, best
supportive care
PP02.37: MESOCLIN: A FRENCH NATIONAL
NETWORK OF EXPERT CENTERS FOR THE
MANAGEMENT OF MPM PATIENTS AND FOR
RESEARCH PROMOTION
Arnaud Scherpereel1, Myriam Locatelli2, Laurent Greillier3 , Jacques Margery4 , David Planchard5, Xavier Dhalluin1,
Françoise Galateau-Sallé6 , Eric Wasielewski1, Françoise Le
Pimpec-Barthes7
Hospital of the University (CHU) of Lille, Lille, FRANCE, 2Hospital of the University (CHU) of Lyon, Lyon, FRANCE, 3Assistance
Publique Hôpitaux de Marseille, Marseille, FRANCE, 4Hôpital
Percy, Paris, FRANCE, 5IGR, Villejuif, FRANCE, 6MESOBANK, Lyon,
FRANCE, 7Inserm U.1162, INSERM U.1162, PARIS, FRANCE
1
tumor with poor prognosis, usually associated with previous
asbestos exposure. Its management, from the diagnosis to a
multimodal treatment, may be complex and tricky. In 2012,
the national French institute against cancer (INCa) funded a
network of expert centers to improve the management of this
tumor with the help of patients’ associations and to stimulate
the research in this field.
Methods: 15 French centers were established according to the
experience of their multimodal team and board in MPM (number of patients per center, publications in the field…). Starting
2016, every case of MPM in France (submitted online through
dedicated software and website) should benefit from diagnostic and/or therapeutic advice by these experts teams. In close
collaboration with partners such as Mesopath, Institut National
de Veille Sanitaire (InVS), asbestos victims associations, French
intergroup of thoracic oncology (IFCT)…, we also aim at stimulating all research studies and clinical trials in MPM, helping the
recruitment of patients and the collection of data and samples
in the MESOBANK project.
Results: Between 2012 and 2014, 1736 (new or pretreated)
MPM patients were managed by the teams of the MESOCLIN
network. It represents about one third of the new patients in
France during the same period of time; 271 out of these 1736
patients were recruited into clinical trials, most of them in academic trials. Full and updated data of the MESOCLIN network
will be presented during the 2016 iMig meeting.
Conclusion: The MESOCLIN network was not fully established
and effective yet till 2016 but, in collaboration with all our
partners, it aims at targeting an exhaustive and comprehensive management of MPM patients in France, with associated
research studies and clinical trials.
Keywords: network, management, mesothelioma, research
Conclusion: Maintenance treatment with gemcitabine is feasible in patients with mesothelioma.
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ABSTRACT BOOK
PP02.39: DOES OPEN ACCESS EXPERT PHONE
TRIAGE BASED ON 2012 IMIG GUIDELINES FOR
VETERANS RECEIVING CARE WITHIN THE VA,
ALTER THERAPY?
Charles J. Siegert1, Pietro M. Fisichella2, Quin Huang3 , Jennifer
Moseley2, Abraham Lebenthal4
Surgery, Brigham and Women’s / Boston VA, West Roxbury,
UNITED STATES OF AMERICA, 2Surgery, VA Boston Healthcare
System, West Roxbury, UNITED STATES OF AMERICA, 3Pathology,
VA Boston Healthcare System, West Roxbury, UNITED STATES OF
AMERICA, 4Surgery, Brigham and Women’s Hospital/ Boston VA,
West Roxbury, UNITED STATES OF AMERICA
1
Objectives: INTRODUCTION. In the United States, Veterans
are comprise less than 7% of the population yet account for
approximately 1/33 of new Malignant Pleural Mesothelioma
(MPM) patients annually. MPM is often recognized as a service
connected disease. The overwhelming majority of Veterans
within VHA are not offered maximal cytoreductive surgery. The
VHA is the largest integrated healthcare system in the United
States caring for 10 million Veterans. The Boston VA Healthcare
System (VABHS) is a Harvard teaching hospital, providing:
regional (New England), in network, high volume, complex
tertiary general thoracic surgical care. Staff surgeons hold
appointments at VABHS and Brigham and Women’s Hospital (BWH), and are members of International Mesothelioma
Program (IMP). In attempt to improve access for Veterans, we
decided to pilot: ‘open access expert phone triage’ nationally.
Utilizing IMiG 2012 guidelines, we attempt to asses feasibility of
phone triage and travel for consultation, impact on therapeutic
recommendations and ultimate in network treatment Veterans
received at VABHS.
Methods: Following iMig 2012, the senior author, a general
thoracic surgeon, specializing in MPM provided open access
phone triage to Veterans eligible for VHA Patients were referred
by multiple sources. After initial screening interview patients
were instructed to setup electronic access to their medical
records at VABHS, and send by overnight mail: disks containing imaging and pathology slides. We reviewed records in a
multidisciplinary setting, independently evaluating source data
(imaging, pathology, etc.). Veterans that appeared to be reasonable candidates for cytoreductive surgery were encouraged
to fly to Boston for further assessment. Veterans that received
appropriate care locally were reassured.
Results: 91 patients attempted to utilize our phone triage, 60
were US Veterans, 16 were where out of state non-veterans that
were excluded, and 14 where international patients seeking
therapeutic guidance (this was offered pro-bono, 12/14 were
non-veterans)). Not all of the 60 veterans interviewed over the
phone had active VHA benefits, of those eligible, 38 patients
from 25 states travelled an average 997 miles to VABHS for
initial surgical consultation. 2 likely operative candidates chose
non operative therapy locally. The majority of the remainder
chose surgical consultation elsewhere.
In 71% (27/38) of patients we examined at the VABHS initial
therapeutic plans were alter based on IMiG guidelines. These
patients were evaluated locally and deemed non surgical candidates, 18/27 were recommended definitive chemotherapy 7/27
were initially offered palliative care, 1/27 radiation and chemotherapy and 1/27 surgical resection. Ultimately, 21 out of the 27
patients (78%) had definitive resection (11 radical pleurectomy/
decortication P/D and 10 extrapleural pneumonectomy EPP). 2
patients were advised to start chemotherapy for extrathoracic
disease rather than initiate a resection. 3 patients chose to not
pursue surgery and chose to continue chemotherapy. One patient surgery was aborted due to unresectable disease following
neoadjuvant therapy for N2 disease.
Conclusion: Open access expert phone triage and travel based
on 2012 IMiG guidelines for Veterans receiving care within the
VHA is feasible for the majority of patients that were encouraged to come for surgical second opinion altering treatment
recommendations and therapy.
Keywords: Veterans, mesothelioma, IMiG guidelines, Veteran
Healthcare System
PP02.41: MINE PROJECT - MESOTHELIOMA
INFORMATION NETWORK IN EUROPE
Giorgio V. Scagliotti1, Egbert Smit2, Carlo Di Pietrantonj3 , Gerald
Schmid-Bindert4 , Marianne Nicolson5, Corrado Magnani6 , David Planchard7, Luis Paz-Ares8 , Silvia Mattone9, Natalia Motas10
Department of Oncology, University of Turin, Torino, ITALY, 2Vu
University Medical Center, Amsterdam, NETHERLANDS, 3ASL
AL, Alessandria, ITALY, 4UMM, Mannheim, GERMANY, 5NHS
Grampian, Aberdeen, UNITED KINGDOM, 6Dep. Translational
Medicine, University Eastern Medicine, Novara, ITALY, 7IGR,
Villejuif, FRANCE, 8FIBH12O, Madrid, SPAIN, 9Oncology Department, Università degli Studi di Torino, Orbassano, ITALY, 10IOB,
Bucharest, ROMANIA
1
Objectives: MINE (Mesothelioma Information Network in
Europe) is a European project funded by CHAFEA (Consumer,
Health, Agriculture and Food Executive Agency). It includes
a partnership of 9 institutions from Italy, France, Germany,
Romania, Spain, The Netherlands and United Kingdom. The
project, through the realization of a European network of MPM
centres, aims to contribute to the advancement of current
knowledge and to the harmonization of diagnostic and therapeutic processes for MPM and to increase awareness of risks
in target populations. MINE will analyze the state of the art of
the guidelines for MPM diagnosis and therapy and promote the
development and harmonization of MM registries and tissue
banks and foster the cooperation among clinical centres so to
provide homogeneous statistics and specific guidelines.
Methods: The advancement of those objectives will be pursued
along three lines: a) update and dissemination of guidelines; b)
information and facilitation of the participation in clinical trials,
and c) availability of biological material for preclinical research
on MM. The methods employed by the project are: surveys of
diagnostic and therapeutic procedures in selected hospitals
on clinical, radiological and pathological procedures for MPM.
Evidence based guidelines on diagnosis and therapy of MPM
are prepared following state of the art methodology. Analysis
of existing MM registries and local database collections and
their interaction with tissue banks will be reviewed. A systematic mapping of the existing biological tissue banks for MM is
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ABSTRACT BOOK
pursued with a survey of pathology units. MINE will generate
user-friendly educational and informative materials to be made
available at large and to be also disseminated through primary
care physicians and advocacy groups to target groups subjects
in selected areas of high relevance.
Results: The expected results of the project will be to create
and disseminate a report describing the current diagnostic
treatment procedures for MPM in Europe with a focus on the
major needs for development on clinical and epidemiological
resources for MPM in Eastern Europe. We will work on improved tissue bank guidelines and on a proposal for harmonization in MPM registration. The work will also focus on asbestos
related risks and the importance on health surveillance.
Conclusion: So far, the project designed and disseminated
the surveys and results are currently analyzed. This analysis
will provide the bases on which the future steps will be built:
gathering the differences among European centres in diagnosis,
treatment, registration and collection of samples that will help
the consortium in creating harmonized and updated guidelines
on the topics covered by the project and it will help in providing
the landscape of the MPM in European countries.
Results: The review of attendance information suggests that
four categories of people requiring support can be identified: 1.
Patients who are newly diagnosed and want clinical information
and support, 2. Patients in a stable condition, who want to live
a ‘normal’ life, 3. Patients with symptomatic and progressive
disease, and 4. The bereaved who struggle with grief and loss.
There are barriers that impact on each category and potentially
affect the uptake of our service. They include: 1. Travel time (up
to 3 hours and distances of more than 100 kms.) 2. Appointments for treatment that need to be kept as well as changes in
performance status that may affect travel capabilities. 3. Some
patients prefer to avoid the confrontation with other patients
and only welcome one-on-one conversation.
Conclusion: While developing our MPM support service we
have identified that patients and carers prefer a personalised
approach to physical and emotional support. In the coming period we will continue to monitor the uptake of our service and at
the same time explore the addition of interactive technologies
to target typical physical symptoms of MPM and the informational needs of patients, carers and the bereaved and translate
this in a personalised approach. 1. Mesothelioma in Australia
2012. 4th Annual Report. Australian Mesothelioma Registry,
2014 2. Van Zandwijk N, Clarke C, Henderson D, et al. Guidelines for the diagnosis and treatment of malignant pleural mesothelioma. Journal of Thoracic Disease. 2013;5(6):E254-E307.
doi:10.3978/j.issn.2072-1439.2013.11.28.
Keywords: support, personalised, patients’, mesothelioma,
PP02.42: PERSONALISED SUPPORT FOR PATIENTS
WITH MALIGNANT PLEURAL MESOTHELIOMA
(MPM) IN NEW SOUTH WALES, AUSTRALIA
Jocelyn Mclean1, Nico Van Zandwijk2
1
Asbestos Diseases Research Institute, Sydney, AUSTRALIA, 2Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA
Objectives: The Australian Mesothelioma Registry reported
641 new cases of MPM in 2014. Of those cases, 185 were from
NSW. 1 The symptom burden of this incurable disease is very
high as is the need for medical and emotional support.2 Data
about the needs of these patients is currently being collected in
an observational study of health related quality-of-life in people
with MPM at the Asbestos Diseases Research Institute (ADRI).
In the meantime, we have started to develop a support service
that will reflect the personal needs of patients, carers and the
bereaved across NSW. Three different groups of individuals
with a request for support were identified: patients receiving
standard (palliative) care, patients who have had radical (combined-modality) treatment, and the bereaved.
Methods: The Australian Mesothelioma Registry reported
641 new cases of MPM in 2014. Of those cases, 185 were from
NSW. 1 The symptom burden of this incurable disease is very
high as is the need for medical and emotional support.2 Data
about the needs of these patients is currently being collected in
an observational study of health related quality-of-life in people
with MPM at the Asbestos Diseases Research Institute (ADRI).
In the meantime, we have started to develop a support service
that will reflect the personal needs of patients, carers and the
bereaved across NSW. Three different groups of individuals
with a request for support were identified: patients receiving
standard (palliative) care, patients who have had radical (combined-modality) treatment, and the bereaved.
PP02.43: SURGICAL CYTOREDUCTION
CHEMOTHERAPY (HITHOC) FOR MALIGNANT
Michael Ried1, Reiner Neu1, Christian Großer2, Tamas Szöke2,
Hans-Stefan Hofmann1
Department of Thoracic Surgery, University Medical Center
Regensburg, Regensburg, GERMANY, 2Department of Thoracic
Surgery, Hospital Barmherzige Brüder Regensburg, Regensburg,
GERMANY
1
Objectives: Combination of surgical cytoreduction and hyperthermic intrathoracic chemotherapy (HITHOC) perfusion is
performed more often for therapy of malignant pleural mesothelioma (MPM) within a multimodality treatment concept. We
describe our perioperative management and clinical experience.
Methods: Between September 2008 and January 2015 a total
of n= 23 patients with MPM were enrolled. Perioperative management, postoperative morbidity and mortality were analyzed.
Next follow-up analysis is scheduled for April 2016.
Results: Included were n= 18 male and n= 5 female patients
with a mean age of 61.7 ± 8.2 years. All patients received
multimodality therapy depending on tumor stage, histology and
their overall condition. Histologic subtype of patients with MPM
was epitheloid (n= 19; 83%) or biphasic (n= 4; 17%). Induction
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ABSTRACT BOOK
chemotherapy with cisplatin/pemetrexed was performed in n=
21 patients (91%). All patients underwent radical surgical cytoreduction with pleurectomy/decortication (P/D; n= 13; 57%),
extended P/D (resection of pericardium and/or diaphragm;
n= 7; 30%) or extrapleural pneumonectomy (EPP; n= 3; 13%)
followed by HITHOC perfusion at 42°C for one hour. HITHOC
was performed with an increasing concentration of cisplatin
(100 mg/m2 BSA n= 7; 150 mg/m2 BSA n= 11; 175 mg/m2 BSA
n= 1) or combination of cisplatin/doxorubicin (175 mg/m2 BSA
/ 65 mg n= 4). Macroscopic complete resection (R0/R1) was
achieved in n= 22 patients (96%). Severe chemotherapy-related complications were not observed. Operative revision was
necessary in n= 2 patients (9%) due to rupture of the diaphragmatic patch. Postoperative renal insufficiency was observed
in n= 3 patients (13%) with no patient requiring temporary
postoperative dialysis (0%). Prolonged bronchopleural fistula
was documented in n= 2 patients (9%) after lungsparing P/D
or eP/D. 30-day mortality was 4.4%, because n= 1 patient died
after EPP. Adjuvant chemotherapy was accomplished in n= 14
patients (61%). Last follow-up analysis in July 2015 showed a
mean recurrence free interval of 23 months and mean overall
survival was 32 months. Taken together, at this time n= 12
patients (52%) were still alive.
Conclusion: Surgical cytoreduction in combination with
HITHOC can be performed with acceptable morbidity and mortality rates in selected patients. Patients should be evaluated
interdisciplinary to determine their eligibility for this multimodality approach. Early clinical results may encourage the use of
additional HITHOC to provide better local tumor control.
Keywords: pleural mesothelioma, hyperthermic intrathoracic
chemotherapy, pleurectomy/decortication, surgical cytoreduction
MPM at clinically IMIG-stage II underwent RP via right-sided
postero-lateral thoracotomy as part of multimodality treatment.
RP was performed as a standardized surgical procedure at our
institution. Additionally, partial resection of the diaphragm and
thymus was performed. Soft tissue foci at the chest wall were
resected. The diode-pumped Nd:YAG Laser LIMAX® 120 (wavelength: 1318 nm, Gebrüder Martin GmbH & Co KG, Tuttlingen,
Germany) and a power output of 100 watts was utilized to
scissor through lung tissue in case of lung infiltration aiming to
accomplish MCR.
Results: With a hand piece and under direct visual control, the
diode-pumped Nd:YAG laser permitted intraparenchymal cut
through lung tissue in the basilar segments. Simultaneously,
the diode-pumped Nd:YAG laser caused tissue vaporization
and coagulation of the resection surface, respectively. The
appearance and functionality especially of the lower lobe could
be preserved. The blood loss could be maintained low (approximately 450 mL). Lung-scarifying procedures could be avoided.
MCR could be achieved at the end of the procedure. Operative
time (4 hours 47 minutes) was acceptable despite the extent of
surgery. No intraoperative side-effects of the laser application
could be observed. Pathological IMIG-stage was found to be IV
(pT4 pN0 (0/17)) due to completely resectable tumor extending
into the soft tissue of the chest wall (two foci) in addition to the
lung infiltration. The patient could be discharged from hospital
at postoperative day 13 after removal of the pleural drainages.
No postoperative complications occurred. He successfully underwent adjuvant radiation of the chest wall and chemotherapy
without any delay.
Conclusion: The diode-pumped Nd:YAG laser might be an
important tool in the surgical armamentarium for parenchyma-sparing, macroscopic complete resections in the treatment
of malignant pleural mesothelioma. However, further experimental investigations and clinical studies are warranted.
Keywords: Malignant pleural mesothelioma, lung-sparing
surgery, diode-pumped Nd:YAG laser
PP02.44: DIODE-PUMPED ND:YAG LASER FOR
LUNG-SPARING SURGICAL TREATMENT OF
MALIGNANT PLEURAL MESOTHELIOMA - FIRST
EXPERIENCE
Servet Bölükbas , Michael Eberlein
1
2
Thoracic Surgery, Helios Klinikum Wuppertal, Wuppertal,
GERMANY, 2Pulmonary, Critical Care And Occupational Medicine, University of Iowa Hospitals and Clinics, Iowa, IA, UNITED
STATES OF AMERICA
1
Objectives: The goal of surgery is macroscopic complete
resection (MCR) for multimodal treatment of malignant pleural
mesothelioma (MPM). Deep infiltration of the basilar segments
of the lung might obviate lung-sparing radical pleuractomy (RP).
Commonly, lung-sacrifying procedures (extrapleural pneumonectomies or lobectomies) are performed in these situations. It
has been demonstrated that high-output Nd:YAG laser precisely
scissor, seal lung tissue and coagulate at the same time for the
treatment of lung metastases. We report our first experience
with diode-pumped Nd:YAG laser intending to avoid lung-sacrifying surgeries for MPM.
Methods: A 53-year-old patient with biopsy-proven epitheloid
PP02.45: PATHOLOGICAL EVALUATION OF THE
VISCERAL PLEURA STAMP IN THE RADICAL
PLEURECTOMY/DECORTICATION FOR MPM
PATIENTS
Masashi Kobayashi, Chihiro Takasaki, Sachiko Kumazawa,
Hironori Ishibashi, Kenichi Okubo
Thoracic Surgery, Tokyo Medical and Dental University, Tokyo,
JAPAN
Objectives: In recent years, procedure of radical pleurectomy/
decortications (P/D) was increased in surgical treatment of
resectable malignant pleural mesothelioma. And macroscopic
complete resection (MCR) in resectable MPM is most important. However, there are no reports such as clinical results of
evaluating providence with visceral pleura side pathologically
in P/D cases. Visceral pleura is microscopically composed of
mesothelial layer, submesothelial layer, external elastic layer,
interstitial layer, and internal elastic layer. We investigated the
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140
ABSTRACT BOOK
dissection plane of decortication of the visceral pleura from the
lung.
Methods: Ten MPM patients who underwent radical pleurectomy/decortication at Tokyo Medical and Dental University
Hospital between April 2010 between October 2015 were
studied. Among these patients, there were 8 epithelioid tumors,
1 biphasic tumor, and 1 desmoplastic tumor, - 4 patients with
stage I MPMs, 2 patients with stage II MPMs, 4 patients with
stage III MPMs. We performed analysis to site of visceral pleural
lesions in radical MPM patients, using the EV& HE staining
to evaluate for ten cases and site of visceral pleural as: Right
upper 5 lesions, middle 6 lesions, lower 7 lesion Left upper 5
lesions, lower 8 lesions, totally 31 lesions.
Results: In all specimens, the growth of tumor cells in the
visceral pleura surface was indicated in diffuse partially or
nodules. Fourteen of 31 lesions with MPM cell invasion directly
to the lung parenchyma epitomized, but all 14 lesions with any
depth of invasion of them were excised from the lung parenchyma. On the other hand, although 17 lesion sites were excised at
lung parenchyma, those lesions were remained intravascular
pleura without invasion of lung parenchyma. Each visceral
pleuron was dissected in lung parenchyma, no relation to depth
of tumor invasion or pleural thickening on all lesions. Furthermore, regardless of extent of invasion, lung parenchyma was
disconnected in the visceral pleura in the vicinity of the site in
the pleural thickening; thin section was cut off in the deep part
of the lung parenchyma. There was no residual tumor in lung
parenchyma for 31 lesions.
Conclusion: In respectable malignant pleural mesothelioma,
visceral pleura dissection plane in the P/D was lung parenchyma.
Keywords: radical pleurectomy/decortication
PP02.46: CLINICAL AND IMMUNOLOGIC
IMPACT OF SURGERY FOR MALIGNANT PLEURAL
MESOTHELIOMA
Vincenzo Ambrogi1, Franco Stella2, Tommaso Claudio Mineo1
Thoracic Surgery, Tor Vergata University, Rome, ITALY, 2Thoracic
Surgery, University of Bologna, Bologna, ITALY
1
Objectives: The aim of the two intentionally-curative surgical
procedures, extra-pleural pneumonectomy and radical pleurectomy/decortication, is mainly focused on the impact on survival.
Unfortunately, this is not significantly improved whatever the
operation. Conversely, the clinical impact of surgery is poorly
investigated. The object of this study is to analyze the effects of
these two operations on immunology, symptoms and quality of
life.
pathology undergoing a non-intentionally curative procedure
(i.e. video-assisted thoracoscopy plus biopsy and possible pleurodesis). The effects on immunitary response were analyzed
with total lymphocyte count, lymphocytes subpopulations
and interleukin-6 and 10, measured pre and postoperatively
(days 1, 7 and 14). Symptoms, function and quality of life were
assessed before surgery and at 3, 6, 12 and 18 months during
the follow-up. Quality of life was tested with Medical Outcomes
Study SF-36 and the St. George’s Respiratory Questionnaire,
administered to the patients at the same intervals.
Results: There was no perioperative mortality in any group.
One patient died 20 days after extrapleural pneumonectomy for
pulmonary embolism. Thirty-day postoperative major morbidity was 45% (14/31) for extrapleural pneumonectomy, 23%
(10/44) for pleurectomy/decortication and 14% (5/35) for the
control group (p=0.04). Median survivals in the extrapleural
pneumonectomy and pleurectomy/decortication groups were
20 and 15 months, respectively (p=0.09), whereas in the control
group was 10 months. Total lymphocyte count and natural killer
subtype significantly decreased in both intentionally-curative
groups, but especially after extrapleural pneumonectomy.
Interleukin-6 and -10 were significantly increased in all groups.
However, intergroup comparison evidenced more elevated
values in the extrapleural pneumonectomy group at postoperative day 7 for both interleukin-6 (p=0.01) and interleukin-10
(p=0.04). The control group showed a faster normalization of
interleukin-10 values when compared to the other procedures.
In the early postoperative period patients undergoing pleurectomy/decortication presented greater symptomatic and functional
improvement. After 1 year patients treated with extrapleural
pneumonectomy generally showed greater improvements.
Nearly all SF-36 domains showed a significant amelioration at 3
months in the pleurectomy/decortication group. After extrapleural pneumonectomy we experienced a slower amelioration
of all domains except the physical component. Thereafter all
domains, physical component included, improved and persisted
significantly better than the preoperative value up to one year.
In the control group the improvements were insignificant. Similar scores were assessed with the George’s questionnaire. After
one year we experienced a progressive worsening of all quality
of life parameters. However more durable values were found in
the extrapleural pneumonectomy group.
Conclusion: Both surgical procedures performed with curative
intent still have a scant impact on overall survival. Extrapleural
pneumonectomy had a greater negative impact on immunological status thus producing a significantly greater risk of
postoperative infection implying a higher morbidity rate. Both
procedures were effective in ameliorating symptoms and quality of life with a more rapid effect for pleurectomy/decortication
but longer for extrapleural pneumonectomy.
Keywords: Surgery, Immunology, Malignant pleural mesothelioma, quality of life
Methods: From 1995 to 2014 a total of 75 with malignant
pleural mesothelioma underwent extrapleural pneumonectomy
(n=31) or pleurectomy-decortication (n=44) with intentionally
curative intent in our two centres. These two groups were also
compared with a control group of 35 patients with the same
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ABSTRACT BOOK
Table 1. Demographics and Perioperative Variables
PP02.47: WHO BENEFITS FROM MACROSCOPIC
COMPLETE RESECTION IN MALIGNANT PLEURAL
MESOTHELIOMA?
Hasan Batirel1, Muzaffer Metintas2, Hale B. Caglar3 , Guntulu
Ak2, Fulden Yumuk4 , Rengin Ahiskali5, Zeynep Bilgi1, Tunc Lacin1, Bedrettin Yildizeli1, Mustafa Yuksel1
Thoracic Surgery, Marmara University Faculty of Medicine,
Istanbul, TURKEY, 2Pulmonary Medicine, Eskisehir Osmangazi
University, Eskisehir, TURKEY, 3Radiation Oncology, Medipol University Faculty of Medicine, Istanbul, TURKEY, 4Internal Medicine,
Div Of Medical Oncology, Marmara University Faculty of Medicine, Istanbul, TURKEY, 5Pathology, Marmara University Faculty
of Medicine, Istanbul, TURKEY
1
Objectives: Macroscopic complete resection (MCR) is the recommended surgical principle in malignant pleural mesothelioma (MPM). The objectives of this study are to analyze whether
MCR contributes to survival in a specific subgroup of patients
and to compare outcomes in patients who underwent PD with
MCR and partial PD.
Methods: Between September 2005 and August 2015, 80
patients underwent PD for MPM in our clinic. All surgeries were
performed through a posterolateral thoracotomy with removal
of 6th rib. Diaphragmatic and pericardial reconstruction were
performed in 19 and 13 patients respectively. If grossly visible
tumor was left behind, the resection was accepted as partial
PD. Patient data were recorded in a prospective database.
Demographic criteria (age, gender), histology, postoperative
morbidity and mortality (90-day), length of hospital stay, pathologic stage, use of neoadjuvant/adjuvant treatment, follow-up,
site of recurrence and survival were recorded. Whole cohort,
patients who had PD with MCR (n=33) or partial PD (n=47)
were evaluated separately. Student t-test, Kaplan-Meier survival
and uni- and multivariate analyses were performed.
Results: Average age was 56 ± 11 (36 females). 51 had
epithelioid histology. Postoperative morbidity occurred in 20
(prolonged air leak in 11). Mortality was seen in 2 patients due
to sepsis and ARDS. Median length of hospital stay was 7.5 ±
3.5 days. Upfront treatment was applied in 23. 70 underwent
adjuvant treatment. Mean follow-up was 18 ± 15 months.
Recurrence occurred in 61 patients (only locoregional [n=50],
locoregional and distant [n=8], only abdomen [n=3]). Overall
median survival was 17.4 months. 2 and 5-year survivals were
35 and 19% respectively. In uni- and multivariate analysis
histology and postoperative N status were significant (0.05 and
0.026 respectively). Patients with epithelioid histology, N0 status and PD with MCR (n=16) had a 2-year survival of 71% and
median survival was not reached. Comparison of patients who
had PD with MCR to partial PD are shown in Table 1. Overall and
2-year survivals were similar between patients who had PD with
MCR or partial PD. PD with
MCR
(n=33)
Partial PD
(n=47)
p value
Age (y), Gender
(Male/Female)
55 ± 11,
14/19
56 ± 10,
30/17
0.5, 0.06
Histology (Epithelioid/Biphasic/Sarcomatoid) (n)
22/10/1
29/15/3
0.68
T1+2/T3+4 (n)
22/11
9/38
<0.001
N (0/1/2/X) (n)
25/1/
5/2
17/0/3/27
<0.001
Upfront treatment (n)
7
16
0.22
Perioperative Morbidity/90-day Mortality
(n)
13/2
7/0
0.013/
0.09
Hospital Stay (d)
9 ± 4.5
6.5 ± 2.2
0.001
Median and 2-year
Survival (months/%)
14.1/39
18.2/33
0.76
Conclusion: MCR does not translate to prolonged survival in
all patients with MPM who undergo PD. Patients with epithelioid histology and N0 status benefit most from this surgical
technique. Efforts should be focused on better preoperative
mediastinal N staging and histologic diagnosis
Keywords: pleurectomy/decortication, Macroscopic Complete
Resection
PP02.48: IS SURGERY FOR MESOTHELIOMA IN
THE UK A DYING MODALITY IN THE MANAGEMENT
OF MALIGNANT MESOTHELIOMA? UPDATE ON
OUR EXPERIENCE
Mohammed Khalil1, Syed Qadri2, Mubarak Chaudhry2,
Alexander Cale2, Mahmoud Loubani2, Michael Cowen2
Cardiothoracic Surgery, Castle Hill Hospital, Cottingham,
UNITED KINGDOM, 2Castle Hill Hospital, Cottingham, UNITED
KINGDOM
1
Objectives: The Mesothelioma and Radical Surgery (MARS)
trial adversely affected surgery for mesothelioma in the UK,
especially extrapleural pneumonectomy (EPP) with significantly
reduced referrals, although we had demonstrated good outcome of EPP in our department. Following iMIG 2012, we have
changed our practice to pleurectomy/decortication (EPD) as a
part of trimodality treatment if they were not fit for extrapleural
pneumonectomy (EPP). We aim to present our result of limited
experience.
Methods: All patients who had any surgical procedure for
malignant mesothelioma cytoreduction except EPP were
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included. Thirty-four patients underwent pleurectomy, extended
pleurectomy/decortication or radical pleurectomy for malignant
mesothelioma from September 2007 to April 2015.
Results: Thirty-three patients underwent extended pleurectomy/decortication. Median age was 65 years with 82% being
male with mean postoperative hospital stay of 11 days. There
was no in-hospital or 30-days mortality. Overall median survival
was 16.5±5 months (CI 6.7-26.2) with longer survival in epitheloid than biphasic mesothelioma.
Conclusion: This update of our limited experience still reflects
poor referral from respiratory physicians for surgery. We here
continue to demonstrate that surgery for mesothelioma as a
part of trimodality treatment is safe with no operative mortality,
acceptable morbidity and better survival.
Keywords: pleurodecortication, trimodailty
PP02.49: THE ANALYSIS OF THE RECURRENCE
OF THE PATIENTS WHO UNDERWENT SURGICAL
RESECTION FOR MALIGNANT PLEURAL
MESOTHELIOMA
Nobuyuki Kondo1, Toru Nakamichi2, Ayumi Kuroda2, Masaki
Hashimoto2, Teruhisa Takuwa2, Seiji Matsumoto2, Kozo
Kuribayashi3 , Tohru Tsujimura4 , Noriaki Tsubota5, Takashi
Nakano6 , Seiki Hasegawa1
Department of Thoracic Surgery, Hyogo College of Medicine,
Nishinomiya, JAPAN, 2Department of Thoracic Surgery, Hyogo College of Medicine, Nisinomiya City, JAPAN, 3Respiratory
Medicine, Hyogo College of Medicine, Nishinomiya Hyogo,
JAPAN, 4Department of Pathology, Hyogo College of Medicine,
Nishinomiya, Hyogo, JAPAN, 5Department of Thoracic Oncology,
Hyogo College of Medicine, Nishinomiya, JAPAN, 6Hyogo College
of Medicine, Nishinomiya, JAPAN
1
Objectives: Currently combined treatment modalities are the
most frequently used approach for malignant pleural mesothelioma (MPM) as a feasible therapeutic strategy. Because surgery
for malignant pleural mesothelioma (MPM) is cytoreductive, not
radical, a certain ratio of recurrence is inevitable after curative-intent operation. Here we analyzed the recurrence after
surgery with induction chemotherapy for MPM patients (n=100).
Methods: From 2004 to 2015, Hyogo College of Medicine MPM
Surgery Program has given induction chemotherapy followed by
surgery with or without radiation therapy to surgical candidates
with histologically confirmed MPM, clinical stage T1-3N0-1M0
disease, performance status 0–1, and no major comorbidity.
Surgery for MPM contains extrapleural pneumonectomy (EPP)
and pleuractomy/decortication (P/D). Surgery for MPM contains
extrapleural pneumonectomy (EPP) and pleuractomy/decortication (P/D).
Results: 100 consecutive patients (81 male and 19 female)
underwent surgical resewction. The median age was 61
years(range, 37-74years). The numbers of pathological stage I/
II/III/IV were 11/19/55/15, respectively. Histological types were
diagnosed as epithelioid (n=89),biphasic (n=8), sarcomatoid
(n=1), othrs (n=2). 93 patients underwent surgical resection
: 58 EPP, 35 P/D, 7 exploratory thoracotomy, respectively.
Macroscopic complete resection was achieved in 87 patients.
Of 100 patients, 64 patients were confirmed the recurrence
within the observation period. The site of recurrence is classified: new lesion in the ipsilateral thoracic cavity (chest wall,
mediastinum and residual lung) were found in 39 (61.0%)/15
(23.4%) in the abdominal cavity/ 3 (4.7%) in subcutaneous or
muscular tissue/ 9 (14.1%) lymph node metastasis/ 12 (19.1%)
in contralateral thoracic cavity/ 3 (4.7%) distant metastasis. 9
cases were expired without confirmation of recurrence of MPM
(14.1%) but revealed recurrence or metastasis after an autopsy.
Recurrence-free survival time was 434 days. In EPP group, recurrence in ipsilateral thorax was (57.1%), whereas recurrence
and metastasis at other distant site were relatively frequent: the
abdominal cavity (4.7%), lymph node metastasis (14.1%), contralateral thoracic cavity (19.1%), and distant metastasis (7.1%).
In contrast, most of recurrence were found in the ipsilateral
thorax in P/D group.
Conclusion: We examined the recurrence in 100 of MPM
patients who underwent surgical resection in combination with
chemotherapy.
Keywords: pleuractomy/decortication, recurrence, extrapleural pneumonectomy
PP02.50: IPSILATERAL PNEUMONECTOMY AFTER
PLEURECTOMY/DECORTICATION IN A PATIENT
WITH MALIGNANT PLEURAL MESOTHELIOMA
Toru Nakamichi, Seiki Hasegawa, Nobuyuki Kondo, Masaki
Hashimoto, Teruhisa Takuwa, Ayumi Kuroda
Nisinomiya City, JAPAN
Objectives: Background: One of the purposes of pleurectomy/
decortication (P/D) in patients with malignant pleural mesothelioma (MPM) is to induce ipsilateral lung re-expansion by
resecting thickened visceral pleura. However, in patients with
highly restrictive lungs, re-expansion failure of decorticated
lungs may directly cause persistent air leakage and intrathoracic infection. Selection of surgical technique and intraoperative
decision whether or not to discard such lungs is highly difficult.
Methods: Case report
Results: A 72-year-old male underwent resection of right
ycT2N0M0 epithelioid MPM after three cycles of preoperative
chemotherapy. Selection of surgical technique was debatable
because the right lung was highly collapsed before and after
video-assisted thoracoscopic pleural biopsy performed three
months before. Trapped lung was speculated as the cause of
restriction rather than tumor invasion to the pulmonary parenchyma. Although P/D was successfully performed, re-expansion
of decorticated right lung was insufficient and air leakage was
massive. Discarding right lung was considered during operation, but was not adopted in expectation of postoperative gradiMig2016.ORG
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ABSTRACT BOOK
ual improvement. On day 3, severe respiratory failure due to
persistent air leakage and massive suppurative secretion from
collapsed right lung developed and the patient was intubated.
On day 10, life-saving right pneumonectomy was required due
to progressive infiltration in the left lung.The patient successfully recovered.
Conclusion: P/D is contraindicated when the diseased lung
is presumably non-reexpandable. Selection between P/D and
extrapleural pneumonectomy is sometimes difficult in case
where reexpansion potential is unclear. Intraoperative decision to discard poorly-expandable lungs after completion of
decortication is also very hard. However, it should be noted that
such lungs are not only non-functional but also life-threatening.
In the present case, discard of the right lung should have been
decided just after completion of decortication.
Results: Of 129 registered patients, 11 were excluded from this
analysis: 2 referred after induction chemotherapy, 6 refused
surgery after registration, and 3 did not undergo induction
chemotherapy. Response to the induction chemotherapy in
the remaining 118 patients were partial response (20 patients,
16.9%), stable disease (87 patients, 73.7%), and progressive
disease (11 patients, 9.3%). A total of 11 patients (9.3% of 118)
could not undergo surgery due to progressive disease (n=10,
8.4% of 118) or serious adverse event of chemotherapy (n=1,
0.8%). 107 underwent surgery (EPP 56, P/D 44, exploratory
thoracotomy 7). Median survival time and 5-year survival for
operated (n=107) and non-operated patients (n=11) were 38.6
and 11.0 months, and 30.3% and 0%, respectively. Surgical
mortality rates at 30 and 90 days were 1.9% (2/107) and 3.7
(4/107), and surgical morbidity was 34.6% (37/107). Keywords: Ipsilateral pneumonectomy, pleurectomy/decortication
PP02.51: INDUCTION CHEMOTHERAPY
FOLLOWED BY SURGERY FOR MALIGNANT
Nobuyuki Kondo1, Toru Nakamichi1, Ayumi Kuroda1, Masaki
Hashimoto1, Teruhisa Takuwa1, Seiji Matsumoto1, Yoshitomo
Okumura2, Taiichiro Otsuki3 , Kozo Kuribayashi3 , Fumihiro
Tanaka4 , Noriaki Tsubota5, Tohru Tsujimura6 , Norihiko
Kamikonya7, Takashi Nakano3 , Seiki Hasegawa1
Nishinomiya, JAPAN, 2Thoracic Surgery, Itami City Hospital,
Itami, JAPAN, 3Respiratory Medicine, Hyogo College of Medicine,
Nishinomiya Hyogo, JAPAN, 4Surgery Ii, University of Occupational and Environmental Health, Kitakyusyu, JAPAN, 5Department
of Thoracic Oncology, Hyogo College of Medicine, Nishinomiya,
JAPAN, 6Department of Pathology, Hyogo College of Medicine,
Nishinomiya, Hyogo, JAPAN, 7Department of Radiation Oncology,
Hyogo College of Medicine, Nishinomiya, JAPAN
1
Objectives: Since any surgery for malignant pleural mesothelioma (MPM) is cytoreductive, not radical, effective chemotherapy is a prerequisite of curative-intent operation. However, it
remains unclear whether chemotherapy should be administered
before or after surgery. Here we report our 11-year experience
with induction chemotherapy followed by surgery.
Methods: Hyogo College of Medicine MPM Surgery Program
has given induction chemotherapy followed by surgery with or
without radiation therapy to all surgical candidates with histologically confirmed MPM, clinical stage T1-3N0-1M0 disease,
performance status 0–1, and no major comorbidity. From
March 2004 to December 2015, 129 patients were intended to
undergo surgical treatment after chemotherapy. Trimodality
treatment with induction chemotherapy followed by extrapleural pneumonectomy (EPP) and hemithoracic 54Gy radiation
therapy has been intended to all patients during 2004 and
2012. After 2012, most patients underwent bimodality treatment with induction chemotherapy followed by pleuractomy/
decortication (P/D) and postoperative chemotherapy.
Conclusion: Approximately 90% of the patients with surgical intent successfully underwent either of EPP or P/D after
induction chemotherapy with acceptable surgical mortality and
morbidity.
Keywords: Surgery, induction chemotherapy
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ABSTRACT BOOK
PP02.52: ANTI-TUMOR EFFECTS OF METFORMIN
AND NUTLIN-3A IN MALIGNANT PLEURAL
MESOTHELIOMA
Yuji Tada1, Takao Morinaga2, Toshio Suzuki3 , Hideaki Shimada4 ,
Koichiro Tatsumi5, Kenzo Hiroshima6 , Masatoshi Tagawa2
Respirology, School of Medicine Chiba University, Chiba,
JAPAN, 2Chiba Cancer Center Research Institite, Chiba,
JAPAN, 3Chiba University, Chiba, JAPAN, 4Department of Surgery,
School of Medicine, Toho University, Tokyo, JAPAN, 5Department
of Respirology, Graduate School of Medicine, Chiba University,
Chiba, JAPAN, 6Tokyo Women`s Medical University, Yachiyo Medical Center, Yachiyo, JAPAN
1
Objectives: Metformin has been widely used as an oral
drug for type 2 diabetes mellitus. Recent reports showed that
metformin exhibited an anti-tumor effects for a variety of
malignancies. However its effect for malignant pleural mesothelioma (MPM) remains unknown. Therefor we investigated its
impact on mesothelioma cell lines in aspects of cell growth and
viability. We also examined anti-tumor effects of metformin in
combination with nutlin-3a (an inhibitor of p53-degradation).
Methods: We examined the anti-proliferative effect of metformin and/ or nutlin-3a in the effect for eight MPM cell lines.
RNA interference was conducted to clarify the relevance of p53
status and the anti-tumor effects of metformin and nutlin-3a.
Results: Metformin suppressed cell growth of MPM cells in a
p53-independent mechanism. Flow cytometric analysis showed
that metformin treatment markedly induced cell cycle arrest at
G2/M phase and nutlin-3a produced G1 cell cycle arrest. The
apoptotic effect of combination of metformin with nutlin- 3a in
MPM involved multiple mechanism and was dependent to the
cell type.
Conclusion: Metformin suppressed growth of MPM cells in a
p53-independent mechanism. Combinatory use of metformin
and nutlin-3aproduced synergetic inhibitory effects for cell
proliferation.
PP02.53: COMPARED HIGH-RESOLUTION WHOLE
GENOME SCREENING OF MESOTHELIOMA AND
BENIGN ASBESTOS PLEURISY
Tunç Tuncel1, Guntulu Ak2, Hasan Veysi Guneş1, Selma
Metintas3 , Irfan Değirmenci1, Muzaffer Metintas2
Department of Medical Biology, Eskisehir Osmangazi University
Medical Faculty, Eskisehir, TURKEY, 2Lung and Pleural Cancers
Research Aand Clinical Center And Medical Faculty Department
of Chest Diseases, Eskisehir Osmangazi University, Eskisehir,
TURKEY, 3Lung and Pleural Cancers Research and Clinical Center
and Medical Faculty Department of Public Health, Eskisehir
Osmangazi University, Eskisehir, TURKEY
1
the body and etiologicialy linked to asbestos exposure. Although
genetic basis of MPM tumors commonly charecterized with
deletions on several cancer related genes like CDKN2A (9p21),
NF2 (22q12), BAP1 (3p21), WT1 (11p13), BCL10 (1p22), majority
of cases exhibits tumor spesific genetic alterations with great
genomic heterogenity. Also pathological effect of asbestos
exposure on this genomic complexity still remains unknown.
In this study we aimed to determine which asbestos related
genomic alterations can lead malignant transformation on bening asbestos related diseases and help us to determine better
diagnosis and new personalized treatment options. To clarify
these issues, we used high resolution genomic approaches to
detect if there is an asbestos related genomic signature which
can intersect in or distinguish between malignant and bening
diseases.
Methods: A total of 55 MPMs and 18 cases with Benign
Asbestos Pleurisy (BAP) as control group were included in the
study. BAP genomes compared with asbestos related-MPM
genomes to find out genetic factors which are spesific to
asbestos damage. Genomic DNAs isolated from pleural tumor
samples for MPM group and pleural tissues from patients with
BAP. Affymetrix CytoScan HD whole genome SNP array was
assessed in these 73 individuals. According to their complex
genomic architecture, three of these MPM patients selected to
whole genome sequencing (Illumina HiSeq platform) for confirmation SNP array findings and further detailed investigation
for novel complex structural alterations. Data implemantation
and Bioinformatic analysis carried with Nexus Copy Number
Discovery Edition 7,5. Through Nexus 7.5, We analyzed copy
number frequencies with combining GISTIC (Genomic Identification of significant targets in cancer) analysis. Compared
CNV analysis between malign and benign group was conducted
with a %40 genomic aberration frequency difference threshold.
Most frequent and significant copy number changes selected in
combination with these analyses.
Results: Deletion of Interferon locus on 9p21.3 genes including
IFNA7, IFNA10, IFNA16, IFNA17, IFNA14 was detected as most
frequent aberrations among all MPM samples (%65) but not
in benign group. Genomic gains are predominantly found in
different parts of 8q, 5p, 7p and 20q and losses are 22q, 10q,
14q, 13q, 4p-4q, 16p. Both losses and gains are detected in 1p,
3p-3q, 15q, 19p-19q in MPMs. MPM genomes exhibits allelic
imbalance on chromosomal regions like 6q, 9p, 10q, 22q and
3p with ≥%50 frequency. Complex structural alterations like
Chromothripsis patterns was detected as a novel finding on
MPM and it is seen mostly affecting on Chromosome 15.
Conclusion: We detected some genetic alterations between
MPM and BAP patients. We think that these alterations may be
important to understand the pathogenesis of the diseases and
some of them can give opportunites to make new diagnostic
and therapeutic researchs. *This study was carried out with the
biological samples collected in the study named as “Network
cooperation for the management of environmental and occupational exposure to mineral fibers induced pulmonary pathologies” which was supported by General Directorate of Health
Researches, Republic of Turkey, Ministry of Health.
Keywords: genetic, mesothelioma, Biology, pathogenesis
Objectives: Malignant Pleural Mesothelioma (MPM) is a highly
aggressive tumour generally originating from serosal linings of
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ABSTRACT BOOK
mesothelioma and could be a therapeutic target. This work
PP02.54: ANALYSIS OF GENE-EXPRESSION
CHANGES IN 3D SPHEROIDS HIGHLIGHTS A
SURVIVAL ROLE FOR ASS1 IN MESOTHELIOMA
Dario Barbone1, Carlo Follo1, Loes Van Dam2, Puthen V. Jitesh3 ,
Shu Dong Zhang4 , William G. Richards5, Raphael Bueno5, Dean
Fennell6 , Courtney Broaddus1
University of California, San Francisco, San Francisco, CA, UNITED STATES OF AMERICA, 2University Medical Center, Utrecht,
Utrecht, NETHERLANDS, 3Sidra Medical and Research Center,
Doha, QATAR, 4Queen’s University of Belfast, Belfast, UNITED KINGDOM, 5Brigham and Women’s Hospital, Boston, MA,
UNITED STATES OF AMERICA, 6University of Leicester, Leicester,
UNITED KINGDOM
was supported by a Mesothelioma Applied Research Foundation
(MARF) grant to DB (A121342) and by the Simmons Mesothelioma Foundation. CF is supported by a Simmons Fellowship in
Mesothelioma Research.
Keywords: spheroids, arginine, ASS1, chemoresistance
1
Objectives: Understanding the mechanisms of chemoresistance of malignant pleural mesothelioma may help identify
novel therapeutic avenues. We have previously studied how the
3D environment confers increased chemoresistance (multicellular resistance) and wondered if this could be understood by
studying the genes differentially expressed in 3D.
Methods: Using Affymetrix arrays, we investigated the
gene-expression differences between monolayers (2D) and 3D
spheroids grown from three mesothelioma cell lines, M28, REN
and VAMT. The differentially expressed genes were then compared to genes shown to be upregulated in human mesothelioma, in two publicly available datasets comprising data from
100 patients. The staining of one differentially expressed gene,
argininosuccinate synthase or ASS1, was measured in two sets
of tumor microarrays, one containing 88 tumor samples and
one containing samples from 88 additional mesotheliomas with
their paired normal tissues. The difference between the ASS1 of
the tumor and the normal matched control was correlated with
patient outcome. RNA interference was used to ablate ASS1 in
3D spheroids to determine its effect on chemoresistance in that
setting.
Results: A total of 209 genes were differentially expressed in
3D (138 up-regulated and 71 down-regulated) compared to 2D.
Of 3 genes initially found to be upregulated in both 3D spheroids and patient tumors, only ASS1 was found to be consistently upregulated, both at the mRNA and protein level. In the
tumor microarray containing samples from 88 mesothelioma
patients, ASS1 expression was found in 90% of samples; only
8 tumors (~10%) showed no ASS1 staining. In the matched
pairs (tumor vs normal tissue control obtained from the same
patients), ASS1 expression was found to be significantly higher
in mesothelioma than in normal tissue. Moreover, the tumors
that showed the highest difference between ASS1 in the tumor
compared to the paired normal tissue (top quartile, n=24)
had significantly shorter survival than those with a smaller
difference (lower 3 quartiles, n=64) (10.6 vs 20.2 months, p=
0.0004). Ablation of ASS1 by RNA interference significantly
reduced the multicellular resistance of the spheroids.
Conclusion: We have shown that ASS1 is expressed in most
mesothelioma tumors and may play a survival role in a 3D setting. ASS1, which catalyzes the penultimate step of the biosynthetic pathway of the essential amino acid arginine, may thereby assist tumor survival. We propose that ASS1 contributes to
the multicellular resistance acquired by mesothelioma cells in
3D, may contribute to a poor clinical outcome of patients with
PP02.55: MYOSIN II-DEPENDENT CELL
CONTRACTILITY DRIVES SPONTANEOUS NODULE
FORMATION OF MESOTHELIOMA CELLS
Julia Tarnoki-Zach1, Dona Greta Isai2, Elod Mehes1, Sandor
Paku3 , Zoltan Neufeld4 , Balazs Hegedus3 , Balazs Dome5,
Andras Czirok1
Department of Biological Physics, Eotvos Lorand University,
Budapest, HUNGARY, 2Department of Anatomy & Cell Biology, University of Kansas School of Medicine, Kansas City, KS,
UNITED STATES OF AMERICA, 3Mta-se Tumor Progression
Research Group, Hungarian Academy of Sciences, Budapest,
HUNGARY, 4School of Mathematics and Physics, University of
Queensland, Brisbane, ACT, AUSTRALIA, 5Division of Thoracic
Surgery, Department of Surgery, Comprehensive Cancer Center,
Medical University of Vienna, Vienna, AUSTRIA
1
Objectives: Despite recent advances in its treatment, malignant pleural mesothelioma (MPM) still has a poor prognosis
with almost all patients succumbing to disease. A pathognomonic feature of MPM is the formation of multiple, macroscopic
pleural tumor nodules, which may pinch off, and contribute to
the local spreading of the disease. In this study we focus on the
role of cell contractility in MPM nodule formation in vitro.
Methods: Several human patient-derived MPM cell lines were
cultured up to three weeks. The time course of nodule formation was observed by videomicroscopy. Actin and beta-catenin
labelled samples were analyzed by confocal laser scanning
microscopy. To interfere with normal actomyosin function, we
utilized Y27632, the rho kinase (ROCK) inhibitor and blebbistatin, an inhibitor of actomyosin crosslinking.
Results: Macroscopic multicellular aggregates develop when
MPM cell lines are cultured at high cell densities. Surprisingly, the nodule-like aggregates do not arise by excessive local
cell proliferation, but by myosin II-driven cell contractility.
Accordingly, nodule formation can be prevented or reversed
by pharmacological inhibitors of myosin II activity. Contractile nodules contain actin cables that can span multiple cells.
Several features of the in vitro MPM nodule development, e.g.
characteristic pattern size and density or speed of appearance
of aggregates, can be explained by a computational model that
assumes uniform and steady intercellular contractile forces
within a monolayer of cells, and a mechanical load-dependent
lifetime of cell-cell contacts.
Conclusion: Our study indicated the presence of multicellular
stress cables, structures that MPM cells can utilize for longrange communication within the mechanically interlinked tumor
nodules. The cellular contractile activity can exert forces that
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can internalize parts of the host tissue environment including
preexisting blood vessels. The demonstrated ability of myosin
II inhibitors to scatter mesothelioma nodules may open novel
combined therapeutic methods.
Keywords: Malignant pleural mesothelioma, actomyosin contractility, multicellular stress cables, computational model
PP02.57: IDENTIFICATION OF CIS- AND TRANSACTING ELEMENTS REGULATING CALRETININ
EXPRESSION IN MESOTHELIOMA CELLS
Jelena Kresoja-Rakic1, Esra Kapaklikaya1, Gabriela Ziltener1,
Damian Dalcher2, Raffaella Santoro2, Brock C. Christensen3 ,
Kevin C. Johnson3 , Beat Schwaller4 , Walter Weder1, Rolf Stahel5,
Emanuela Felley-Bosco1
University of Zurich, Laboratory of Molecular Oncology, Division
of Thoracic Surgery, Zurich, SWITZERLAND, 2Institute of Veterinary Biochemistry and Molecular Biology, University of Zurich,
Zurich, SWITZERLAND, 3Departments of Epidemiology, Pharmacology and Toxicology and Community and Family, Hanover,
NH, UNITED STATES OF AMERICA, 4Department of Medicine and
Anatomy, Department of Medicine and Anatomy, University of Fribourg, Fribourg, SWITZERLAND, 5Clinic For Oncology, University
Hospital Zurich, Zurich, SWITZERLAND
1
PP02.56: POST-TRANSCRIPTIONAL REGULATION
OF CALRETININ EXPRESSION
Jelena Kresoja-Rakic1, Merve Sulemani1, Michaela B.
Kirschner2, Glen Reid3 , Steven Kao3 , Beat Schwaller4 , Rolf Stahel5, Walter Weder1, Emanuela Felley-Bosco1
University of Zurich, Laboratory of Molecular Oncology, Division
of Thoracic Surgery, Zurich, SWITZERLAND, 2University Hospital
Zurich, Division of Thoracic Surgery, Zurich, SWITZERLAND, 3Asbestos Diseases Research Institute, Sydney, AUSTRALIA, 4Department of Medicine and Anatomy, Department of Medicine and
Anatomy, University of Fribourg, Fribourg, SWITZERLAND, 5Clinic
For Oncology, University Hospital Zurich, Zurich, SWITZERLAND
Objectives: Calretinin (CALB2) is a diagnostic marker for
epithelioid mesothelioma. It is also a prognostic marker since
patients with tumors expressing high calretinin levels have
better overall survival. Silencing of calretinin decreases viability
of epithelioid mesothelioma cells. Our aim was to elucidate
mechanisms regulating calretinin expression in mesothelioma.
Objectives: Calretinin (CALB2) is a diagnostic and prognostic marker in malignant pleural mesothelioma (MPM).
The CALB2 3’UTR contains several putative microRNA target
sites. Our aim was to investigate the role of the CALB2 3’-UTR
in the post-transcriptional regulation of calretinin expression in
MPM.
Methods: To investigate human calretinin (CALB2) promoter,
~1kb of genomic sequence surrounding the transcription start
site (TSS) +1 was analyzed using luciferase reporter pGL3-basic
vector. Transcriptional activity of 5’-deletion CALB2 promoter
constructs in mesothelioma cells was measured via dual luciferase assay. Mutant constructs were created by site-directed
mutagenesis. Electrophoretic mobility shift (EMSA) assay and
chromatin immunoprecipitation (ChIP) assay were used to show
binding of functional transcription factors (TF). To demonstrate
cell-cycle regulated calretinin expression, mesothelioma cells
were synchronized using double thymidine treatment followed
by nocodazole treatment.
1
Methods: Using the pmirGLO Dual-Luciferase expression vector, the complete CALB2 3‘-UTR fragment was inserted 3‘ of the
firefly luciferase gene. Activity of the CALB2 3‘-UTR was quantified after transient transfection into ACC-MESO-4, ZL55 and
ONE-58 MPM cells. In silico analysis was employed to predict
potential microRNAs targeting the CALB2 3’-UTR. Subsequently,
luciferase activity and calretinin expression were evaluated
after the transfection-mediated overexpression of the predicted
microRNAs. In addition, calretinin protein, assessed by immunohistochemistry, and miR-30 expression, were investigated in
a cohort of MPM patients (N=48).
Results: The addition of the CALB2 3’-UTR significantly downregulated the luciferase activity in the three tested MPM cell
lines. Analysis of the CALB2 3’-UTR using the TargetScan online
database predicted target sites for the miR-30 family members,
miR-9 and miR-384. Transient delivery of a miR-30e mimic
into CALB 3’UTR stably-transfected ACC-MESO-4 cells resulted in an even further decrease of the activity of the luciferase
reporter as well as a decrease in calretinin protein expression.
Finally, expression of miR-30e was found to negatively correlate with the calretinin expression in a cohort of MPM patient
samples.
Conclusion: Our data shows for the first time the role of miR30e in the post-transcriptional negative regulation of calretinin
expression via interaction with its 3’-UTR.
Keywords: calretinin, mesothelioma, 3’-UTR, microRNA
Results: Analysis of calretinin transcript and protein suggested
a control at the mRNA level. Treatment with 5-aza-2’-deoxycytidine and analysis of TCGA data indicated that promoter
methylation is not likely to be involved. Therefore, we investigated the CALB2 promoter by analyzing ~1kb of genomic
sequence surrounding the transcription start site (TSS) +1 using
promoter reporter assay Deletion analysis of the CALB2 proximal promoter showed that sequence spanning the -161/+80bp
region sustained transcriptional activity. Site-directed analysis identified important cis-regulatory elements within this
-161/+80bp CALB2 promoter. EMSA and ChIP assays confirmed
binding of NRF-1 and E2F2 to the CALB2 promoter and siRNA
knockdown of NRF-1 led to decreased expression of calretinin.
Cell synchronization experiment showed that calretinin expression was cell cycle regulated with a peak of expression at G1/S
phase.
Conclusion: Our study identified the transcription factors
NRF-1 and E2F2 to bind to the human CALB2 promoter and
we demonstrated cell cycle-dependent regulation of calretinin
expression in mesothelioma cells providing the first insight into
the regulation of CALB2 expression in mesothelioma cells.
Keywords: calretinin, promoter, NRF-1, E2F, cell-cycle
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ABSTRACT BOOK
PP02.58: DECREASED PROLIFERATION AND CELL
MIGRATION OF PRIMARY MESOTHELIAL CELLS
FROM CALRETININ-DEFICIENT (CR-/-) MICE
Walter Blum1, Emanuela Felley-Bosco2, Laszlo Pecze3 , Beat
Schwaller1
Department of Medicine and Anatomy, Department of Medicine and Anatomy, University of Fribourg, Fribourg, SWITZERLAND, 2Department of Molecular Oncology, University Hospital
Zurich, Zurich, SWITZERLAND, 3Medicine, University of Fribourg,
Fribourg, SWITZERLAND
1
Objectives: Calretinin (CR; human gene symbol: CALB2) is one
of the most sensitive and specific markers for the diagnosis of
malignant pleural mesothelioma (MPM). Interestingly also reactive human mesothelial cells (proliferating, non-transformed
cells) show strong expression of CR. CR’s exact function in
mesothelioma formation and its putative molecular involvement
is still unknown. In order to investigate CR’s possible role in the
reactive mesothelium and in the presumed first steps of tumor
development, we investigated the role of CR in mouse primary
mesothelial cells (prMC). Our objective was to compare prMC
from wildtype (WT) and from CR knockout (CR-/-) mice concerning growth, proliferation, cell cycle length and migration
and the effect of artificially (via-lentivirus) up-regulating CR.
Methods: prMC from WT and CR-/- mice were analyzed for
morphological, proliferative and migratory characteristics. Fluorescent cell-cycle indicators (Fucci) allowed to determining cell
cycle parameters. Efficient over-expression of CR and NLS-CR
(nuclear localization signal) was achieved using lentiviral-mediated transduction of prMC. Mouse embryos at an age of 14.5
and 16.5 days were collected and investigated by IHC for CR
expression.
Results: Analysis of the mesothelium from WT and CR-/- mice
showed no noticeable macroscopic and histologic differences.
PrMC derived from both genotypes were isolated and grown in
vitro. Their in vitro morphology was “cobblestone-like”, they
expressed the mesothelial markers mesothelin, cytokeratin and
vimentin and TEM analysis revealed the presence of microvilli.
In CR-/- cells we observed a statistically significantly decreased
proliferation rate. By up-regulating CR in prMC (WT and CR-/-)
the proliferation rate and the wound closure (mobility) rate was
increased; the up-regulation also induced a more pronounced
epithelial morphology. The prMC originating from CR-/- animals
had a prolonged G1 phase, but an unchanged S/G2/M phase. In
the scratch assay the wound closure time of CR-deficient prMC
was significantly longer. Artificial over-expression of CR and
NLS-CR in prMC (WT and CR-/-) led to a change in cell morphology, an increased proliferation rate and mobility. WT cells
closed the wound in the scratch assay much faster and this was
due to a combined effect of increased proliferation rate and
higher cell mobility of the cells at the wound borders. Immunohistological analysis of embryos (E14.5 and 16.5) showed transient CR expression in cells of the embryonic connective tissue
(mesenchyme) and in mesothelial precursor cells surrounding
the developing lung. Differentiated mesothelial cells (flat morphology) showed no longer immunostaining for CR.
Conclusion: The absence of CR during the embryonic development in CR-/- mice results in long-lasting changes in mesothelial cell characteristics evidenced by differences in vitro in the
proliferation and mobility of prMC from WT and CR-/- mice. We
make the hypothesis that CR plays an important role during the
normal development of mesothelial cells in vivo. Understanding
mechanistically the involvement of CR in normal mesothelial
cells and in reactive mesothelium is expected to help to understand the process of mesotheliomagenesis. Knowledge about
the affected signaling pathways leading to increased proliferation and cell migration might lead to the identification of novel
attractive targets for mesothelioma therapy besides directly
targeting the expression of CR.
Keywords: Calretinin, primary mesothelial cells, Mesothelium
PP02.59: KCNMA1 IS TARGETED BY MIR-175P AND MODULATES CELL MIGRATION IN
Yuen Yee Cheng1, Casey M. Wright1, Michaela B. Kirschner2,
Marissa Williams1, Kadir H. Sarun1, J J. Edelman3 , Michael P.
Vallely3 , Brian C. Mccaughan4 , Sonja Klebe5, Nico Van Zandwijk1, Ruby C. Lin6 , Glen Reid1
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA, 2Division of Thoracic Surgery, University Hospital Zurich, Zurich, SWITZERLAND, 3The Baird Institute and Faculty of Medicine,
The University of Sydney, Sydney, NSW, AUSTRALIA, 4Sydney
Cardiothoracic Surgeons, RPA Medical Centre, Sydney, NSW,
AUSTRALIA, 5Department of Anatomical Pathology, Flinders Medical Centre, Adelaide, SA, AUSTRALIA, 6Cardiothoracic Genomics,
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA
1
Objectives: Mesothelioma has poor prognosis with little
therapeutic progress, thus identification of new therapeutic
targets for MPM is urgently needed. We and others have shown
that microRNAs play an important role in mesothelioma biology
and have potential as therapeutic agents. Here we attempt
to identify dysregulated microRNAs with functional roles by:
utilising publicly available gene expression datasets on MPM,
in combination with our transcriptomics studies; validating
candidate targets in our relatively large biobank collection of
MPM tumours and normal pleural samples; and investigating
the functional roles of these candidates.
Methods: Candidate targets were identified bioinformatically
by systematic interrogation of mRNA-microRNA differential
gene expression correlations using Gene Set Enrichment Analysis. Candidates (mRNA, microRNA and protein) were validated
using RT-qPCR and immunofluorescent assays in mesothelioma
and normal mesothelium samples. Functional significance of
candidate genes was confirmed by a number of assays including SYBR green proliferation assay, colony formation assay,
cell cycle analysis, apoptosis assay (annexin V and PI staining),
migration assay and agarose spot invasion assay.
Results: We identified enrichment of target binding sites for
the miR-17 and miR-30 families in both MPM tumours and cell
lines. RT-qPCR revealed that members of both families were
significantly down-regulated in MPM tumours and cell lines.
Interestingly, lower expression of miR-17-5p (P = 0.022) and
miR-20a-5p (P = 0.026) was clearly associated with epitheliiMig2016.ORG
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ABSTRACT BOOK
oid histology. We interrogated the predicted targets of these
differentially expressed microRNA families in MPM cell lines,
and identified KCa1.1, a calcium-activated potassium channel
subunit alpha 1 encoded by the KCNMA1gene, as a target of
miR-17-5p. KCa1.1 was overexpressed in MPM cells compared
to the immortalised mesothelial line MeT-5A, and was also
up-regulated in patient tumour samples compared to normal
mesothelium. Transfection of MPM cells with a miR-17-5p
mimic or KCNMA1-specific siRNAs reduced mRNA expression of
KCa1.1 and inhibited MPM cell migration. Similarly, treatment
with paxilline, a small molecule inhibitor of KCa1.1, resulted in
suppression of MPM cell migration.
Conclusion: These functional data implicating KCa1.1 in MPM
cell migration support our integrative approach using MPM
gene expression datasets to identify novel and potentially
druggable targets.
Keywords: mesothelioma, miR-17-5p, therapeutic targets,
KCNMA1, KCal.1, calcium-activated potassium channel subunit
alpha 1
PP02.60: ENGINEERED, LIGHT-CONTROLLED
GROWTH FACTOR RECEPTORS FOR
MESOTHELIOMA RESEARCH
Karin Schelch1, Alvaro Ingles-Prieto2, Eva Reichhart2, Stephanie
Kainrath2, Mir A. Hoda3 , Walter Berger1, Harald Janovjak2, Michael Grusch1
Department of Medicine I, Institute of Cancer Research, Medical
University of Vienna, Vienna, AUSTRIA, 2Institute of Science and
Technology (IST) Austria, Klosterneuburg, AUSTRIA, 3Medical
University of Vienna, Vienna, AUSTRIA
1
Objectives: In the field of optogenetics researchers use
genetically encoded signaling molecules that can be activated
or inactivated by light. Our group focuses on the role of growth
factors and their receptors for the malignant behaviour of mesothelioma cells. Our aim was therefore to generate synthetic
growth factor receptors that can be activated by light and allow
to control growth factor-associated signal transduction pathways with superior spatiotemporal precision.
Methods: To generate RTKs that can be optically activated
(Opto-RTKs), intracellular domains of mammalian RTKs were
fused to light-sensitive protein domains of the light-oxygen voltage (LOV) family from various species. The resulting chimeric
receptors were tested for light-dependent activation of signal
transduction by reporter gene assays, immunoblotting, and
various cell biological tests assessing DNA synthesis, epithelial
mesenchymal transition (EMT), and sprout formation.
Results: Three different LOV domains were identified that were
capable of inducing light-dependent dimerisation and activation
of signal transduction when fused to the intracellular domain
of murine fibroblast growth factor receptor 1 (mFGFR1). Similar
results were obtained for additional RTKs including human
EGFR, RET or c-Met. Light-induced activation of Opto-mFGFR1
enabled control of the MAPK, PI3K and PLCγ pathways. Signal
activation could be spatially confined to illuminated regions
of cell cultures and signals rapidly subsided after cessation
of illumination. Functionally, light could replace FGF2 for the
induction of cell proliferation and EMT in mesothelioma cells. In
endothelial cells, light could replace VEGF for the stimulation of
angiogenic sprouting.
Conclusion: Our optogenetic approach enables fine-tuned,
light-mediated control of growth factor receptors kown to be
important for mesothelioma growth. Opto-RTKs will be valuable
tools for a number of applications in mesothelioma research
including dissection of signalling dynamics with increased spatiotemporal precision in single cell experiments, targeting signal
activation to specific cell populations in co-culture systems
and experimental animals, and facilitation of drug screening
procedures.
Keywords: optogenetic, signal transduction, growth factor
receptor, receptor tyrosine kinase
PP02.61: PRECLINICAL INVESTIGATION OF THE
THERAPEUTIC POTENTIAL OF NINTEDANIB IN
Viktoria Laszlo1, Judit Ozsvar1, Thomas Klikovits1, Dora
Lakatos2, Mir A. Hoda1, Tamas Garay3 , Walter Berger4 , Michael
Grusch4 , Walter Klepetko1, Frank Hilberg5, Balazs Dome1,
Balazs Hegedus1
Surgery, Medical University Vienna, Vienna, AUSTRIA, 2Department of Biological Physics, Eotvos University, Budapest, HUNGARY, 32nd Department of Pathology, Semmelweis University,
Budapest, HUNGARY, 4Department of Internal Medicine, Medical
University Vienna, Vienna, AUSTRIA, 5Boehringer Ingelheim
Austria, Vienna, AUSTRIA
1
Objectives: Malignant pleural mesothelioma (MPM) is a
devastating malignancy with still rising incidence worldwide. Its
aggressive biological behavior and therapy resistance result in
a median overall survival (OS) of 9 to 17 months only. Currently,
platinum-based chemotherapy in combination with antifolate
agents is the standard front-line therapy for MPM and to date
no molecularly targeted therapeutic approaches have been
approved in the clinics. Nintedanib is an indolinone derivative
that has been demonstrated to efficiently inhibit the activity of
VEGFR, PDGFR and FGFR tyrosine kinase isoforms and thus to
be capable to suppress angiogenesis and tumor growth. Here,
we report the antitumor activity of nintedanib in MPM.
Methods: 21 MPM cell lines were treated with nintedanib
and SRB assays were performed to determine the IC50 values
for each cell line. 4 sensitive cell models were selected for
further in vitro analysis: BrdU, TUNEL and clonogenic assays
were performed to investigate the impact of the drug on the
proliferation, apoptosis and colony formation capacity of MPM
cells, respectively. The migratory activity of MPM cells was
analyzed with 2D videomicroscopy. The downstream signaling of the target receptors was investigated by Western blot
analysis. Drug interactions with cisplatin were assessed in the
p31 MPM cell line and in its cisplatin-resistant subline (p31cis)
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by using the CalcuSyn software. The in vivo anti-MPM activity of
nintedanib was studied in an orthotopic human MPM xenograft
model in SCID mice. The effect of nintedanib as single agent at
a dose of 50 mg/kg daily, administered intraperitoneally and in
combination with the cisplatin/pemetrexed regimen, was investigated. Survival, total tumor weight and microvessel density
were evaluated
Results: Nintedanib exerted a growth inhibitory effect on MPM
cell lines in both short- and long-term viability assays. The inhibition of proliferation was observed in all MPM cell models analyzed, whereas significant apoptosis induction was only found
in half of them. Migratory activity strongly decreased upon nintedanib treatment. Down-regulation of Erk1/2 phosphorylation
was evident within 10 min of treatment and was present even
after 24h. Nintedanib, however, had no inhibitory effect on the
activation of Akt or S6. Additive, but no synergistic effect on cell
viability was detected in the p31 and p31cis MPM cells when
nintedanib was combined with cisplatin. In vivo, nintedanib
significantly prolonged the survival of mice (vs. control, log-rank
test, p=0.0008) and inhibited tumor growth as a single agent
(p=0.003 vs control), as well as in combination with cisplatin
and pemetrexed (0.005 vs cisplatin-pemetrexed). Moreover, nintedanib inhibited angiogenesis in the xenograft tumors.
Conclusion: Our data suggest that nintedanib exerts antitumor
activity in MPM both in vitro and in vivo and thus may represent
a promising novel therapeutic option in this malignancy
Keywords: angiogenesis, targeted therapy, Nintedanib
content in comparison with normal rats. Primary tumors and
metastases were fixed, embedded in paraffin and analysed for
histopathology and immunohistochemistry.
Results: In vitro, the M5-T2 cell line differed from the three
others by a low saturation density (1.1 x 105 / cm2), low mobility and a myofibroblast-like morphology. In vivo, the tumor
was mainly restricted to the omentum without metastases to
normal tissues. Conversely, F4-T2, F5-T1 and M5-T1 cell lines
shared in common a high saturation density (> 2 x 105 / cm2),
a high mobility, a propensity to produce metastasis to normal
tissues and positivity for vimentin in IHC. Among these cells,
M5-T1 presented the shortest doubling time, highest saturation density and was the only cell line producing spheroids in
culture. It also exhibited the highest expression of Fra-1, which
has been associated with cell migration in both rat and human
mesothelioma. In vivo, M5-T1 tumor was also characterized by
considerable tumor heterogeneity and a very high mitotic index.
Analyses of chemokine content in peritoneal fluids revealed that
M5-T1 and F5-T1 presented elevated levels of MCP-1, relative
to controls, while F4-T2 and M5-T2 did not present any change
in the content of any of the eight chemokines. F5-T1 tumor
differed from M5-T1 by an additional elevation of MCP-3, MIP-1
α, and IP-10.
Conclusion: These four experimental rat tumor models represent interesting new tools for basic research on tumor microenvironment and oncoimmunology, with potential prospects for
the evaluation of new therapeutic strategies for MM.
Keywords: Cell lines, Rat tumor model, Vimentin, Chemokines
PP02.62: COMPARATIVE ANALYSIS OF 4
EXPERIMENTAL MESOTHELIOMAS IN F344 RATS:
A PRELIMINARY STUDY OF THEIR TUMOR BIOLOGY
FEATURES
Joëlle Nader1, Myriam Robard2, Marc Grégoire1, Daniel L.
Pouliquen1
UMR 892 INSERM / 6299 CNRS, Nantes, FRANCE, 2Plateforme
MicroPICell, SFR F. Bonamy, Nantes, FRANCE
1
PP02.63: THBS2, A NOVEL GENE INVOLVED IN
THE MALIGNANT PROGRESSION OF PLEURAL
MESOTHELIOMA
Elisa Barone1, Stefan J. Marciniak2, Doris M. Rassl3 , Julia
Knight3 , James Wason4 , Luciano Mutti5, Alessandra Bonotti6 ,
Rudy Foddis6 , Alfonso Cristaudo6 , Ombretta Melaiu1, Giovanni
Giangreco1, Federica Gemignani1, Stefano Landi1
Objectives: Malignant mesothelioma (MM) develops in complex microenvironments where molecular interactions between
tumor cells and the immune system of the host play a crucial
role. Given the diversity of biologic situations found among
MM, in-depth study of the parameters involved might lead to
formulation of new principles in tumor biology as well as new
therapeutic strategies.
Department of Biology, University of Pisa, Pisa, ITALY, 2Cambridge Institute For Medical Research (cimr), University of
Cambridge, Cambridge, UNITED KINGDOM, 3Department of
Pathology, Papworth Hospital NHS Foundation Trust, Cambridge,
UNITED KINGDOM, 4MRC Biostatistics Unit, Cambridge, UNITED
KINGDOM, 5School of Environment and Life Sciences, University
of Salford, Manchester, UNITED KINGDOM, 6School of Medicine
and Surgery, University of Pisa, Section of Occupational Medicine, Pisa, ITALY
Methods: The cell lines used in this study were established
from Fischer F344 rats, 378 to 392 days after induction with
a single intraperitoneal inoculation of crocidolite fibers (UICC
analytical sample, Neyco). The growth pattern of each cell line
(doubling time, saturation density) was determined in culture
in 6-well plates, and expression profiles of genes of interest
determined by RT-qPCR. Fifteen to thirty five days after orthotopic (i.p.) injection of tumor cells into syngeneic rats, peritoneal
fluids and plasmas were collected for analyses of GRO alpha,
Eotaxin, IP-10, MIP-1 α, MIP-2, MCP-1, MCP-3 and Rantes
Objectives: The identification of novel diagnostic/prognostic biomarkers and therapeutic targets for Malignant Pleural
Mesothelioma (MPM) is of extreme importance. Following
previous studies, we identified genes up-regulated and potentially involved in the carcinogenetic process of MPM. Among
them, THBS2 was found up-regulated in MPM tissues and in
Mero-14, Mero-25, and Istmes2 MPM cell lines. Aims: to evaluate the functional role of THBS2 in MPM cell lines and its use as
a prognostic biomarker.
1
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ABSTRACT BOOK
Methods: In order to investigate the role of THBS2 in the
carcinogenesis of MPM, we employed RNA interference techniques. Following transient transfection, we performed a phenotypic screening through the Sulphorhodamine-b, the Colony
Formation, the Wound-Healing, and the Caspase luminescence
assays. For evaluating the protein expression of THBS2 as
prognostic biomarker we performed immuno-histochemistry
on Tissue Microarray (TMA) composed by 135 cases of MPM
and we correlated the staining scores with the patients’ overall
survival.
Results: THBS2-silencing caused a statistical significant
increase of apoptosis (+30% in Mero-14; p-value= 0.032), a
reduced proliferation (-41% in IstMes2; p-value = 0.044), and
a decreased clonogenicity (-66% in Mero14 cells; p-value =
0.020). No significant association between protein expression
and overall survival was found.
Conclusion: Present results suggest a potential of targeting THBS2 as a novel therapeutic approach for the treatment of
MPM, whereas data from TMA do not support the use of THBS2
as a prognostic biomarker.
Keywords: THBS2, silencing, novel target, mesothelioma
PP02.64: GROWTH FACTOR-INDUCED
MORPHOLOGY AND EXPRESSION CHANGES IN
CELL MODELS REFLECTING THE HISTOLOGICAL
MESOTHELIOMA SUBTYPES
treatment with FGF2 and EGF induced phenotypical changes
reminiscent of EMT. Changes in cell shape were accompanied
by scattering, increased migration, growth and invasiveness
and required signaling along the mitogen-activated protein kinase (MAPK) pathway. Inhibition of the fibroblast growth factor
receptors (FGFR) or the MAPK axis could prevent these changes
and, in MPM cell lines with sarcomatoid morphology, reverse
scattering and induce a more epithelioid morphology. Gene and
microRNA expression analyses demonstrated an overlap with
previously established EMT markers such as CDH1 or VIM, but
also identified several novel potential markers such as MMP1,
ESM1, ETV4, PDL1, ITGA6 or BDKRB2. Blockage of the FGFR
or the MAPK pathway resulted in the opposite regulation of
these genes. Inhibition of MMP1 but not of ESM1 or ETV4 via
siRNAs or pharmacological inhibitors prevented FGF2-induced
scattering and invasiveness. In unsupervised clustering, the
gene expression profiles of solvent- or cytokine-treated cells
were associated with those of epithelioid and sarcomatoid
MPM, respectively. Pathway enrichment analysis of the targets
of altered microRNAs (fold change >10) as well as differentially
expressed genes (fold change >3) after FGF2 treatment showed
that the regulated genes are assigned to categories such as
transcriptional misregulation in cancer, MAPK, Wnt and Hippo
pathway, interaction with extracellular matrix, focal adhesions
and tight junctions.
Conclusion: These findings enhance our understanding of
the morphological and behavioral plasticity of mesothelioma
cells and the link to the MPM histological subtypes and their
influence on patient outcome.
Keywords: Epithelial-mesenchymal transition, MAPK Signals,
MPM histological subtypes
Karin Schelch1, Christina Wagner2, Ruby Lin3 , Mir A. Hoda2,
Balazs Hegedus2, Balazs Dome2, Walter Berger2, Glen Reid3 , Michael Grusch2
University of Sydney, Asbestos Diseases Research Institute,
Sydney, AUSTRALIA, 2Medical University of Vienna, Vienna,
AUSTRIA, 3Asbestos Diseases Research Institute, Sydney, AUSTRALIA
1
Objectives: The three main histological subtypes of malignant
pleural mesothelioma (MPM), epithelioid, sarcomatoid and
biphasic are characterized by differences in aggressiveness
and patient prognosis. However, the mechanisms and causes responsible for the different cell morphologies are poorly
understood. Epithelial-mesenchymal transition (EMT) has been
implicated in progression and chemoresistance of numerous
tumors but its role in MPM is not well characterized. The aim of
this study was to analyze growth factor-induced morphological
and phenotypical changes in MPM cell lines and the associated gene expression and signal transduction cascades in more
detail.
Methods: Morphological and behavioral changes of cell
models treated with cytokines and inhibitors or transfected
with siRNAs were analyzed by morphometry, immunoblotting,
migration, invasion and soft agar assays. Alterations in gene or
microRNA expression were evaluated via qPCR and array hybridization. Pathway enrichment analysis was based on KEGG.
Results: In several cell models established from biphasic MPM,
PP02.65: THE H3K27ME3 DEMETHYLASE
KDM6B AS AN EPIGENETIC REGULATOR OF ERΒ
EXPRESSION IN HUMAN MALIGNANT PLEURAL
MESOTHELIOMA
Arcangela G. Manente1, Giulia Pinton1, Luca Pavesi1, Daniela
Tavian2, Puthen V. Jithesh3 , Dean Fennell4 , Stefan Nilsson5,
Laura Moro1
Novara, ITALY, 2Cellular Biochemistry and Molecular Biology,
Catholic University of the Sacred Heart, Milano, ITALY, 3Sidra
Medical and Research Center, Doha, QATAR, 4Cancer Studies,
University of Leicester, Leicester, UNITED KINGDOM, 5Biosciences
and Nutrition, Karolinska Institutet, Huddinge, SWEDEN
1
Objectives: To determine the correlation between the demethylase KDM6B and the estrogen receptor β (ERβ) expression in
human malignant pleural mesothelioma (MPM).
Methods: We have evaluated the correlation between KDM6B
and ERβ expression in MPM tumor samples and derived cell
lines grown in normoxic and hypoxic conditions.
Results: In this study, we report a strong positive correlation
between high expression of the H3K27 demethylase KDM6B
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and the ERβ coding genes in MPM tumor samples. Furthermore, we demonstrate a role for KDM6B in the control of ERβ
expression in MPM derived cell lines, independent from the
expressed levels of the methyltransferase EZH2. Parallel induction of KDM6B and ERβ occurs in estrogen receptor negative
cells from biphasic MPM, grown in chronic hypoxia or packed
in spheroids, where the presence of a hypoxic core is confirmed
by HIF2 immunofluorescence staining. Real-time and Western
blot analyses confirm the HIF2-dependent hypoxic induction
of KDM6B and the KDM6B-dependent expression of ERβ. The
activation of KDM6B in hypoxia is confirmed by reduced histone
H3K27 tri-methylation. Importantly, the expression of ERβ
and the less aggressive phenotype, acquired in hypoxia, are
maintained when cells return to normoxia if treated with the
selective ERβ agonist KB9520, even though HIF2 and KDM6B
decrease to basal levels.
Conclusion: The possibility to reverse the more aggressive biphasic cell phenotype by targeting ERβ with a selective agonist
could represent a new strategy to effectively treat this histological subtype of MPM.
Keywords: ERbeta, hypoxia, KDM6B, Malignant pleural mesothelioma
PP02.66: COMBINATION OF NUTLIN-3A AND
HSP90 INHIBITORS PRODUCES SYNERGISTIC
CYTOTOXICITY ON MESOTHELIOMA WITH THE
WILD-TYPE P53
Masatoshi Tagawa1, Shinya Okamoto1, Takao Morinaga1,
Masato Shingyoji2, Ikuo Sekine3 , Toshio Suzuki4 , Yuji Tada4 ,
Koichiro Tatsumi4 , Hideaki Shimada5, Kenzo Hiroshima6
Chiba Cancer Center Research Institite, Chiba, JAPAN, 2Division
of Respirology, Chiba Cancer Center, Chiba, JAPAN, 3Department
of Medical Oncology, University of Tsukuba, Tsukuba, JAPAN, 4Department of Respirology, Graduate School of Medicine, Chiba
University, Chiba, JAPAN,5Department of Surgery, School of Medicine, Toho University, Tokyo, JAPAN, 6Department of Pathology,
Tokyo Women’s Medical University, Yachiyo, JAPAN
1
Objectives: A majority of malignant mesothelioma lacks the
INK4A/ARF locus which contains the p14ARF and p16INK4A genes
but possesses the wild-type p53 genotype. The genetic defect
activates MDM2 that facilitates p53 degradation processes, and
consequently induces a functional loss of p53 activities together
with uninhibited cell cycle progression. Activation of the endogenous p53 pathways by inhibiting MDM2 can be crucial for cell
death of mesothelioma. We therefore investigated a possible
therapeutic strategy of the p53 activation with an inhibitors for
MDM2 and MDM4, a MDM2-like molecule involved in suppression of p53-mediated transactivation.
Methods: We examined cytotoxicity of nutlin-3a which blocked
the MDM2-p53 interaction and subsequently suppressed
proteasome-mediated p53 degradation, and that of heat shock
protein 90 (HSP90) inhibitors which repressed p53-inactivating
MDM4 and receptor-type tyrosine kinases. Cytotoxicity was
assessed with a colorimetric assay and the CalcuSyn software,
and cell cycle was analyzed with flow cytometry. Expression
levels of p53-associated proteins were examined with Western
blot analysis.
Results: Nutlin-3a produced cytotoxicity on mesothelioma
with the wild-type p53 but not with mutated p53, whereas
HSP90 inhibitors, 17-AAG and 17-DMAG, achieved cytotoxicity
in a p53-independent manner. Cells treated with nutlin-3a
increased sub-G1 fractions, and showed phosphorylation of p53
and activation of the p53 down-stream pathways. HSP90 inhibitors-treated cells exhibited up-regulation of p53 expression and
down-regulation of AKT phosphorylation although knock-down
of p53 with the si-RNA did not influence the cytotoxicity. Combination of nutlin-3a with HSP90 inhibitors produced synergistic
cytotoxicity with further increased sub-G1 fractions through
augmented p53 expression levels and enhanced caspase
cleavages but not through down-regulated AKP phosphorylation. Nutlin-3a produced synergistic cytotoxic activities with
NSC207895, a specific MDM4 inhibitor, but not with MK-2206,
an AKT inhibitor. The combinatory use of nutlin-3a with HSP90
inhibitors achieved suppression of tumor growth greater than a
single agent in an orthotopic mouse model.
Conclusion: Nutlin-3a and HSP90 inhibitors produced synergistic cytotoxic effects on mesothelioma through inhibiting both
MDM2 and MDM4 functions. Activation of the endogenous p53
by inhibiting the MDM2-mediated p53 degradation processes
and intensifying p53-mediated transcriptional activation due to
MDM4 suppression is favorable for cell death of mesothelioma
bearing the wild-type p53 genotype.
Keywords: HSP90, MDM4, p53, MDM2
PP02.67: SHED SYNDECAN-1 ALTERS
ANGIOGENESIS IN MALIGNANT MESOTHELIOMA
Ghazal Heidari-Hamedani1, Angelika Schmalzl1, Tünde
Szatmari1, Muzaffer Metintas2, Anders Hjerpe1, Katalin Dobra1
KI, Stockholm, SWEDEN, 2Osmangazi University, Eskisehir,
TURKEY
1
Objectives: Angiogenesis is important in mesothelioma
progression, and so far anti-angiogenic therapies have not
shown significant improvement in patients’ survival, highlighting the further need of novel treatment options. Syndecan-1 is
a transmembrane heparan sulfate proteoglycan that acts as a
regulatory co-receptor in different cellular processes including angiogenesis. The angiogenesis regulatory mechanism of
syndecan-1 is known to be through shedding of extracellular
domain of syndecan-1 into body fluids through matrix metalloproteinases (MMPs), and interacting with growth factors
including VEGF. Also SDC1 activates integrins and insulin like
growth factor-1 receptor (IGF-1R). There is a unique sequence
on syndecan-1 extracellular domain known as synstatin that
regulates the signaling pathways involved in angiogenesis. The
expression of syndecan-1 is low on MPM cells and it has been
shown that decrease of syndecan-1 deteriorates the prognosis.
Methods: Syndecan-1 was over-expressed in a human MPM
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cell line and conditioned mediums from syndecan-1 over-expressing cells and mock controls were collected. Expression of
soluble angiogenic factors was measured using Proteome Profiler Array. In order to see if proteins modulated by syndecan-1
overexpression affect endothelial cell proliferation, endothelial
cells were treated with conditioned mediums from syndecan-1
overexpressing cells and controls. Cell proliferation, tube
formation and chemotaxis were assessed using WST1 proliferation assay, 3D tube formation and transwell assays. MMP-7 was
silenced by siRNA to inhibit the shedding. Soluble syndecan-1
and VEGF levels were measured using ELISA in pleural effusions from mesothelioma patients (n=39) or in vitro.
Results: A number of angiogenesis-related proteins were
altered by syndecan-1, including both pro-angiogenic Angiopoietin-1 (Fold change ± SD: 0.65± 0.07), FGF-4 (0.8±0.02),
HGF (1.33±0.07), NRG1-β1 (1.35±0.08), and anti-angiogenic
proteins TSP-1 (0.8±0.02), TIMP-1 (0.89±0.01) and controversial pro/anti-angiogenic proteins such as TGF-β1 (1.35±0.01).
These factors collectively led to inhibition of endothelial cell
proliferation and tube formation. There was no significant
change in endothelial cells chemotactic migration in presence
of SDC1 overexpressing cells conditioned medium compared
to controls. We found a significantly fair correlation between
VEGF and shed SDC1 levels in these patients (r = 0.4; p <0.05).
Median survival time of mesothelioma patients with VEGF level
<2.125 ng/mL was 14 months (n= 11), compared to 6 months in
patients with higher VEGF levels (n=19; hazard ratio: 2.83, 95%
CI: 1.26 to 6.37).
Conclusion: Syndecan-1 overexpression on MPM cells can
change angiogenicity by affecting the growth factors gradient
and inhibiting endothelial cells proliferation and tube formation.
Combination of shed syndecan-1 and VEGF could be a better
prognostic evaluation in mesothelioma patients.
Keywords: angiogenesis, mesothelioma, Shed SDC1
PP02.68: UTILISING MICRORNAS TO
SENSITISE MESOTHELIOMA TO CISPLATIN AND
GEMCITABINE
by seeding 10,000 cells per well in a 96-well round-bottom
suspension culture plate. MPM cells were grown as (2D)
monolayers using standard cell culturing methods. MicroRNA
gene expression in 2D & 3D cell culture was profiled using
TaqMan Low Density Arrays and validated using RT-qPCR and
digital PCR. Drug cytotoxicity to cisplatin and gemcitabine was
investigated in both 2D and 3D cultures. Candidate microRNA
mimics were transfected in both cultures individually to test for
involvement in drug resistance.
Results: We confirmed that MPM cells grown as spheroids are
more resistant to cisplatin and gemcitabine when compared to
MPM cells grown in 2D cultures. Immunofluorescence studies
showed a gradient of hypoxia from the centre of the spheroids
where high Hif1α expression is observed (Fig.1). We also
identified significant up-regulation of miR-210-3p, miR-378a-3p,
miR-195-5p and miR-146b-5p, and down-regulation of miR320b and miR-1225b-5p in 3D spheroids. Transfecting MPM
cells in 2D culture with a miR-210-3p mimic resulted in
increased drug resistance.
Conclusion: We observed that MPM spheroids are more
resistant to cisplatin and gemcitabine than 2D cultures. Hif1α is
highly expressed in the spheroids and all of the significantly differentially expressed microRNAs listed above are involved in the
downstream cascade of Hif1α regulatory pathway. Increasing
the levels of miR-210 using a mimic resulted in increased resistance of MPM cells to chemotherapy, suggesting this microRNA
plays a role in drug resistance observed in MPM.
Keywords: Hif1a, mesothelioma, 3D tumour spheroids, microRNA
Yuen Yee Cheng1, Kadir Sarun1, Michaela B. Kirschner2, Nico
Van Zandwijk1, Glen Reid1, Ruby C. Lin1
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA, 2University Hospital Zurich, Division of Thoracic Surgery,
Zurich, SWITZERLAND
1
aggressive asbestos-related thoracic cancer. Chemotherapy
is an important palliative treatment option but almost every
patient will be confronted with recurrence of disease and drug
resistance. We have shown that microRNAs are involved in
drug response in MPM cells. To better understand the potential
role of microRNAs in drug resistance and sensitisation in MPM
we have used monolayer (2D) and spheroid (3D) cell culture
models.
Methods: Tumour spheroids (a 3D in vitro model) were grown
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PP02.69: TRABECTEDIN IS ACTIVE AS SINGLE
AGENT AND SYNERGIZES WITH CHEMOTHERAPY
AND BCL-2 INHIBITION IN MALIGNANT PLEURAL
MESOTHELIOMA
Mir A. Hoda , Yawen Dong , Christine Pirker , Thomas Klikovits2, Karin Schelch3 , Petra Heffeter1, Kushtrim Kryeziu1, Viktoria
Laszlo2, Balazs Hegedus2, Balazs Dome2, Walter Klepetko2,
Michael Grusch1, Walter Berger1
1
2
1
Institute of Cancer Research, Medical University of Vienna, Vienna, AUSTRIA, 2Division of Thoracic Surgery, Medical University of
Vienna, Vienna, AUSTRIA, 3Asbestos Diseases Research Institute,
Sydney, AUSTRALIA
1
Objectives: Malignant pleural mesothelioma (MPM) is
characterized by widespread resistance to systemic therapy.
Trabectedin is an antineoplastic agent that has been approved
for the treatment of advanced soft tissue sarcoma and ovarian
cancer. In this preclinical study we evaluated the antineoplastic
potential of trabectedin as a single agent and in combination
with cisplatin and bcl-2 inhibition in human MPM.
Methods: Activity of trabectedin alone and in combination was
established in an extended panel of MPM cell lines (N=6) and
primary cell cultures from MPM (N=13) and non-malignant tissues samples (N=2) using chemosensitivity, migration, spheroid
growth, cell cycle distribution and cell death assays. Trabectedin activity in vivo was evaluated in an orthotopic xenograft
model.
Results: Trabectedin exerted a dose-dependent cytotoxic effect
in all MPM cell cultures in vitro when growing as adherent
monolayers or non-adherent spheroids with IC50 values ≤ 10
nM. Non-malignant mesothelial cells were significantly less
responsive. This strong anti-mesothelioma activity was based
on cell cycle perturbation and apoptosis induction synergistically enhanced by cisplatin. Comparison of gene expression
signatures indicated an inverse correlation between trabectedin
response and bcl-2 expression. Accordingly, bcl-2 inhibitors
(obatoxlax, ABT-199) markedly synergized with trabectedin paralleled by deregulated expression of the bcl-2 family members
bim, bax, Mcl-1 and bcl-xL. Trabectedin exerted significant antitumor activity against an intraperitoneal MPM xenograft model.
Conclusion: These data suggest that trabectedin exerts strong
activity in MPM and synergizes with chemotherapy and experimental bcl-2 inhibitors. Thus, it represents a promising new
therapeutic option for MPM.
Keywords: novel therapeutics, synergism, trabectedin
PP02.70: LIVE CELLS MESOTHELIOMA BIOBANK
TO EXPLORE MECHANISMS OF TUMOR
PROGRESSION
Emanuela Felley-Bosco1, Gabriela Ziltener2, Isabelle Opitz3 ,
Walter Weder3 , Rolf Stahel4
Laboratory of Molecular Oncology/thoracic Surgery, Zürich
University Hospital, Zürich, SWITZERLAND, 2University of Zurich,
Laboratory of Molecular Oncology, Division of Thoracic Surgery,
Zurich, SWITZERLAND, 3Division of Thoracic Surgery, University
Hospital Zurich, Zurich, SWITZERLAND, 4Clinic For Oncology,
University Hospital Zurich, Zurich, SWITZERLAND
1
Objectives: We recently observed that loss of epithelioid phenotype occurring in some of the patients at tumor progression
is associated with worst outcome. In order to carry out functional investigations, primary cultures were established. Our aims
were to secure a live cell biobank and obtain proof of principle
that primary cultures chemoresistant models, mimicking tumor
progression observed in the patient, can be obtained in vitro,
providing a useful tool to investigate underlying mechanisms.
Methods: Primary mesothelioma cultures were established
from 210 patients between 2007 and 2014. Some primary
cultures were obtained from the same patient at different times:
diagnosis, at surgery after cisplatin/pemetrexed therapy and
recurrence. Cultures were characterized by analysis of mesothelioma markers using Western blot and gene expression and
compared to original tumor. A primary culture from a chemo
naïve patient was exposed to increasing doses of cisplatin/
pemetrexed during 3 months and the resulting in vitro treated
cells were compared to control in a cytotoxicity assay and by
selected gene profiling.
Results: Three hundred twenty one primary cultures were
established and expanded to collect RNA, protein and frozen
stocks. Eighty six cultures could be grown in complete absence
of serum and 3% oxygen and some of them have been used to
investigate developmental pathways, which are rapidly lost in
serum containing medium and growth in 20% oxygen. Selected
cultures from a same patient were further characterized for the
expression of calretinin, mesothelin, N-cadherin, podoplanin,
YAP, survivin and found to reflect tumor of origin. Development
of chemoresistance toward cisplatin/pemetrexed was observed
in the in vitro treated culture compared to control untreated
cultures grown in parallel. Selected gene expression revealed a
profile similar to parental cultures established before and after
chemotherapy.
Conclusion: The establishment of a large live cell mesothelioma biobank contributes to the international efforts aimed at
improving the handling of this disease since they can be used
to test novel therapeutic approaches. In addition, they may
help understanding mechanisms underlying tumor progression
observed in vivo.
Keywords: chemoresistance, mesothelioma live cell biobank,
tumor progression
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PP02.71: ANALYSIS OF NOVEL RHOA MUTATIONS
Assunta De Rienzo1, Antonios Sideris2, Daniele Sciaranghella1,
Nhien Dao1, William G. Richards1, Raphael Bueno1
Surgery, Brigham and Women’s Hospital and Harvard Medical
School, Boston, MA, UNITED STATES OF AMERICA, 2Surgery, Division of Thoracic Surgery, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, UNITED STATES OF AMERICA
1
Objectives: Malignant Pleural Mesothelioma (MPM) is a rare
asbestos-induced cancer with poor prognosis (median survival
~4-12 months). Treatment is hampered by the lack of early
detection strategies and effective novel therapeutic approaches. We found that the RHO -family GTPase RHOAis a target of
mutations in MPM (Cancer Res., 2016). RHOA resides on 3p21.3
near BAP1 (3p21.1), a gene frequently deleted in MPM. The
objective of this study was to define the role of RHOA as a novel
putative driver gene in MPM. RHOA regulates the actin cytoskeleton during stress fiber and focal adhesion formation, and
contributes to cancer signaling cascades. It has been implicated as an oncogene in multiple hematologic and solid cancers,
such as angioimmunoblastic T-cell lymphoma and diffuse
gastric cancer.
Methods: Ten MPM and matched normal genomic DNA
samples were analyzed by whole-genome sequencing using a
Complete Genomics platform and novel potential driver genes
were identified. RHOA was further investigated in 147 additional
MPM cases by targeted re-sequencing. Ten MPM cell lines were
selected for functional studies. To determine the biological
impact of the novel mutations on their protein products, the
mutant variants were systematically introduced into established
MPM cell lines. The impact of mutated RHOA was assessed
by in vitro assays. To further analyze the properties of the mutated protein, downstream targets were selected and evaluated
by immunoblotting.
Results: Analyses of a total of 157 MPM identified three novel
somatic mutations (2%) in RHOA. Microarray analysis revealed
that RHOA expression levels were significantly associated with
overall survival indicating an adverse effect of elevated expression of this gene. Immunoblotting analyses showed expression
of RHOA at different levels in all 10 MPM cell lines. Pharmacological and genetic inhibition of RHOA appeared to alter the
migratory ability of MPM cell lines.
Conclusion: RHOA was found to be mutated in 2% of MPMs.
Preliminary functional analyses suggest a potential role
of RHOA in the tumorigenesis of MPM. Assays evaluating cell
motility, migratory ability, invasiveness and anchorage-independent growth of the transduced MPM cell lines are in progress
to further characterize the role of mutated RHOA in MPM. This
study may lead to novel anti-tumor interventions and patient
stratification criteria for future trials.
Keywords: Malignant pleural mesothelioma, RHOA
PP02.72: CELL ASSAYS AND SAXS INDICATE
THAT BAMLET IS A POTENTIAL TREATMENT FOR
CHEMOTHERAPY-RESISTANT MESOTHELIOMA
Emma M. Rath1, Yuen Yee Cheng2, Amanda L. Hudson3 , Chris
Weir3 , Kadir Sarun2, Anders P. Håkansson4 , Viive Howell3 ,
Robert B. Knott5, Anthony P. Duff5, Guo J. Liu5, Glen Reid2, W B.
Church1
Faculty of Pharmacy, University of Sydney, Sydney, NSW,
AUSTRALIA, 2Asbestos Diseases Research Institute, Sydney,
NSW, AUSTRALIA, 3Bill Walsh Translational Cancer Research
Laboratory, Kolling Institute of Medical Research, Sydney, NSW,
AUSTRALIA, 4Experimental Infection Medicine, Lund University,
Malmö, SWEDEN, 5Australian Nuclear Science and Technology
Organisation, Lucas Heights, NSW, AUSTRALIA
1
Objectives: BAMLET and related HAMLET-like (Human
Alpha-lactalbumin Made Lethal to Tumours) complexes have
demonstrated broad-spectrum anti-cancer activity in vitro to
over 50 cancer cell lines at concentrations that do not harm
some primary and immortalised non-cancer cells. In vivo experiments carried out for HAMLET and BAMLET in humans, rats
and mice for bladder cancer, colon cancer and glioblastoma
demonstrated anti-tumour efficacy and non-toxicity. The HAMLET family of compounds was discovered during studies on the
properties of human milk. Their structure is a novel protein-lipid
structure consisting of an aggregation of partially unfolded protein making up the majority of the compound’s mass, with fatty
acid molecules bound in the hydrophobic core. The cytotoxicity
of HAMLET-like compounds has not previously been investigated for mesotheliomas. This study tested BAMLET compounds
on many mesothelioma cell lines and a few non-cancer cell
lines from human and rat. The compound structures were also
investigated by small-angle X-ray scattering (SAXS).
Methods: Multiple BAMLET compounds were created from
bovine alpha-lactalbumin and beta-lactoglobulin proteins with
varying amounts of oleic acid. Sybr and Alamar Blue cell death
assays were performed to investigate BAMLET and chemotherapy (cisplatin, pemetrexed, gemcitabine, and vinorelbine)
toxicity towards chemo-sensitive and chemo-resistant mesothelioma (human MM05, MSTO, REN, H28, H226, H2452, H2052,
VMC20, VMC23, VMC33, and VMC40, and rat II-45, II-45-CisR,
-PemR, -GemR, -VLBR, and -ComboR) and non-cancer cell
lines (human fibroblasts and MeT5A, and rat 4/4RM.4). To
investigate the role of the active agent oleic acid in its structure as a function of the amount of oleic acid incorporated,
SAXS was performed on multiple BAMLET compounds using a
Bruker NanoStar II and the Australian Synchrotron SAXS/WAXS
beamline.
Results: In vitro, BAMLET killed all 11 human epithelial-like
and biphasic mesothelioma (IC50 = 33±2 µM) and rat mesothelioma cell lines (IC50 = 30±2 µM) at similar concentrations. This concentration range was not toxic to some primary
non-cancer human cells. Cisplatin and pemetrexed killed the
mesothelioma cell lines at the same dose ranges that they killed
some non-cancer cells (cisplatin IC50 = 5.9±2.8 µM). Chemo-resistant rat mesothelioma cells were just as sensitive to
BAMLET as the chemo-sensitive cells (IC50 = 35±17 µM). The
BAMLET structures as revealed by SAXS are well-correlated
with the amount of incorporated oleic acid and are correlated
with the toxicity results of the cell death assays.
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Conclusion: The results show that BAMLET may be an effective and non-toxic treatment option for mesothelioma patients
whose disease no longer responds to chemotherapy and thus
who currently do not have a drug treatment option.
Keywords: mesothelioma, BAMLET, chemotherapy resistance,
SAXS
Transfection of 1nM miR-137 sensitised MPM cell to both cisplatin and gemcitabine. Pathway enrichment analysis revealed
a number of potential targets of miR-137, including YB-1, which
plays a role in MPM cell proliferation. In vitro experiments
showed miR-137 mimic transfection significantly down-regulated YB-1 mRNA (p < 0.05) and protein, which was attributed to
its role as a growth suppressor.
Conclusion: miR-137 acts as a tumour suppressor in MPM
and is significantly down-regulated in both MPM cell lines and
tumour samples. Additionally, miR-137 suppresses the expression of YB-1, which in part explains its tumour suppressive
properties.
PP02.73: TUMOUR SUPPRESSOR MICRORNA-1373P TARGETS ONCO-PROTEIN YB-1 IN MALIGNANT
Keywords: microRNA-137-3p, mesothelioma, therapeutic
targets, YB-1
Thomas Johnson1, Yuen Yee Cheng1, Marissa Williams1, Brian
C. Mccaughan2, Sonja Klebe3 , Nico Van Zandwijk1, Ruby C. Lin4 ,
Glen Reid1
Asbestos Diseases Research Institute, Sydney, NSW, AUSTRALIA, 2Sydney Cardiothoracic Surgeons, RPA Medical Centre,
Sydney, NSW, AUSTRALIA, 3Department of Anatomical Pathology,
Flinders Medical Centre, Adelaide, SA, AUSTRALIA, 4Cardiothoracic Genomics, Asbestos Diseases Research Institute, Sydney,
NSW, AUSTRALIA
1
Objectives: Deletion on a number of chromosomal regions is a
common characteristic observed in malignant pleural mesothelioma (MPM), notably within regions 9p21-22 and 1p21-22.
Specifically, studies have shown that the deletion of region
1p21-22 occurs in approximately 74-85% of MPM cases. Since
this region has been implicated in tumour suppression, we set
out to investigate this phenomenon. MicroRNA-137 (miR-137)
resides within this region and has been shown to modulate
tumour suppression in a number of cancers including breast,
lung and gastric cancer. However, its role in MPM is not yet
clear. Here we investigate whether miR-137 can act as a tumour
suppressor and propose the therapeutic potential of miR-137 in
MPM. We also determine the regulatory nature of miR-137 on
the onco-protein YB-1.
Methods: Gene expression of miR-137 was determined in
10 MPM cell lines, one normal mesothelial line, 125 Formalin-Fixed Paraffin-Embedded (FFPE) MPM tissue samples and
24 normal mesothelial samples using TaqMan probes. Cell proliferation and colony formation assays were conducted following transfection with miR-137 or control mimics to determine
the effect of miR-137 on MPM cell growth. microRNA-137 mimic
and control transfection was also used to test drug sensitisation. Pathway enrichment analyses based on predicted target
gene lists of miR-137 extracted from TargetScan and miRDB
respectively were determined to explore the effect of miR-137 at
a systems level. Validation of miR-137 gene targets by RT-qPCR
and Western Blot were carried out.
Results: miR-137 was significantly down-regulated in four
MPM cell lines (p < 0.01). It is also significantly down-regulated
in FFPE MPM samples compared to the normal mesothelial
samples (p < 0.0001). Proliferation assays revealed growth
inhibition of miR-137 in all ten MPM cell lines, albeit to variable
degrees. Four cell lines exhibited statistically significant growth
retardation (p < 0.05). Colony formation assay results further
supported the suppressive role of miR-137 in MPM proliferation.
PP02.74: IFN REGULATORY FACTOR 9 PLAY KEY
ROLE IN MESOTHELIOMA GROWTH
Yidan D. Zhao1, Licun Wu1, Hana Z. Yun1, Ming Tsao2, Marc De
Perrot1
Thoracic Surgery Labs, University Health Network, Toronto, ON,
CANADA, 2Pathology, Tgh, University Health Network, Toronto,
ON, CANADA
1
Objectives: Interferons (IFNs) are a family of cytokines that
potently demonstrate antitumor, antiviral, immunomodulatory
and antiproliferative activities. Although the complex mechanisms of action remain unclear, IFNs are used the treatment for
a limited number of malignancies, such as melanoma, hairy cell
leukemia, and non-Hodgkins lymphoma. IFN regulatory factor
9 (IRF9) is an IFN regulatory factor that mediates signaling by
type I IFNs (IFNα and IFNγ). IRF9-RNA interference (RNAi) has
a key role completely inhibited the antiproliferative activity, but
its possible roles in mesothelioma are not established. In the
present study, we investigated the effect of host genetic deletion of IRF9 in the mouse on the growth of mesothelioma.
Methods: We developed an asbestos induced mesothelioma
in C57 B/6 mouse in the abdominal cavity 9-14 months after
peritoneal introduction of chrysotile and crocidolite asbestos.
We have also cultured cells derived from these tumors and
established Z1P3 cell line with a variety of epithelioid features
and doubling times around 72 -98 hours. The Z1P3 cells were
then given a dermal injection in both IRF9-deficient and wild
type mice.
Results: The MSLN-imunostaining positive tumors were strikingly similar to epithelioid-like tumor, which resemble to those
occurring in man with regard to histologic features and growth
patterns. In the Z1P3 cells injected groups, tumor growth
were significantly accelerated strongly in IRF9-deficient mice
compared with wild-type (wt) mice.Our results showed that host
IRF9 expression was critical to support an antitumor immune
response and to restrict tumor growth.
Conclusion: In conclusion, we have established a mesothelioma-like Z1P3 cell line and our study shows that in an animal
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innate immunity-related model genetically deleted of an IRF9,
tumor growth are accelerated, suggesting a complex role of
inflammation in host–tumor interaction.
Keywords: asbestos, tumor growth, IFN regulatory factor 9,
MSLN positive
PP02.76: ‘ARE THE PSYCHOLOGICAL NEEDS OF
PATIENTS WITH MESOTHELIOMA THE SAME AS
THOSE WITH ADVANCED LUNG CANCER?’
Hannah Ball
Oxford Pleural Unit, Churchill Hospital, Oxford University Hospitals NHS Foundation Trust, LE, UNITED KINGDOM
PP02.75: PRO-ONCOGENIC EFFECTS OF
PATHOGENIC FIBRES ON MESOTHELIAL CELLS
Joaquin Zacarias-Cabeza1, Tatyana Chernova1, Fiona A. Murphy1, Sara Galavotti1, Xiao-Ming Sun1, Ian R. Powley1, Stefano
Grosso1, Jonathan Bennett2, Apostolos Nakas2, Martin Bushell1,
Anne E. Willis1, Marion Macfarlane1
Hospital, UHL NHS Trust, Leicester, UNITED KINGDOM
1
Objectives: Malignant mesothelioma (MM) is an aggressive,
fatal tumour of the pleura or peritoneum and strongly related to
asbestos exposure. Malignant pleural mesothelioma (MPM) is
the most common and occurs with a latency of up to 40 years.
Long pathogenic fibres fail to clear through the lymph system
and are retained in the pleura of the exposed individuals. During
the long latency period, mesothelial cells remain exposed to
paracrine signalling from the inflammatory cells recruited to
the fibre-retaining areas of the pleura, as well as directly to
fibres. However, the mechanism of malignant transformation of
mesothelial cells is not well understood.
Methods: To examine the effects of paracrine pro-inflammatory factors and cyto- and geno-toxic effects of pathogenic fibres
we used an in-vitro cellular model in which normal untransformed mesothelial cells were exposed to conditioned media
from fibre-activated macrophages or to pathogenic fibres.
Molecular and functional readouts included changes in gene
and protein expression as well as epigenetic changes.
Results: We show that paracrine factors from fibre-activated
macrophages increased proliferation rate and migration ability
in normal mesothelial cells. Moreover, cell death induced by
H2O2 in mesothelial cell culture was reduced by pro-survival
paracrine factors released from fibre-activated macrophages.
Additionally, direct exposure of normal mesothelial cells to
pathogenic fibres with a high-aspect ratio induced changes in
tumour suppressor genes favouring malignant transformation.
Conclusion: Direct exposure of mesothelial cells to fibres
as well as exposure to a pro-inflammatory microenvironment
contribute to the development of pathogenic changes in target
mesothelial cells.
Keywords: Malignant Mesothelioma, Pathogenic Fibres,
Pro-oncogenic, Pro-Inflammatory Factors
Objectives: Mesothelioma is a devastating disease characterised by a poor prognosis, high symptom burden and lack
of effective treatment. Psychological distress which adversely
affects a person’s experience of cancer has been shown to be
highly prevalent in patients with mesothelioma. Historically,
care for those with mesothelioma has been provided by the
existing infrastructure and services in place for lung cancer
and with assumptions having been made that the evidence
guiding the supportive care needs for lung cancer is relevant to
those with mesothelioma. This poster presents the findings of a
systematic literature review with the objective of answering the
question ‘Are the psychological needs of patients with mesothelioma the same as those with advanced lung cancer?’, which
was submitted as the authors dissertation as part of her MSc.
Methods: An electronic search of the databases MEDLINE,
CINAHL, PsycARTICLES, Psychology and Behavioural Sciences
Collection, PsycINFO, and the Cochrane Library of Systematic
Reviews was run. Grey literature was identified and relevant reference lists searched. Studies meeting a predefined inclusion
criterion were read and critically appraised for quality. Data
relating to psychological experiences was extracted which was
then synthesised narratively and through a process of Meta
ethnography.
Results: 17 studies were included in the review. Critical
appraisal identified methodological or reporting weaknesses
across 15 of the studies. Common themes identified across
the studies created 10 key concepts. These were uncertainty,
normality, hope/hopelessness, stigma/blame/guilt, family/carer
concern, physical symptoms, experience of diagnosis, iatrogenic distress, financial/legal and death and dying. Key similarities
and differences were identified between the mesothelioma and
lung cancer evidence.
Conclusion: Conclusions include that there is limited research
exploring the lived experiences of those with mesothelioma and
lung cancer, with the majority of them having methodological
and/or reporting concerns compromising the findings/conclusions made. However, reoccurring themes in the evidence was
found suggesting a number of areas where the psychological
experience of mesothelioma differs from that of advanced lung
cancer. These findings warrant more research to explore further
and if proven, the need for the provision of specialist mesothelioma care services is affirmed. Specialist nurses should
be regularly screening for psychological distress and adapting
supportive care accordingly to meet these needs.
Keywords: Systematic review, nursing, psychological distress,
Supportive care
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PP02.77: “AROUND AUSTRALIA IN 80 DAYS”
ARD/MPM is a small speciality that is competing with a wide
range of other health diseases and therefore has diminished priority status within many health professionals scope of learning.
Judy Rafferty
The vast variances in practices/resources within the country
Cancer Services/Nurse Educator, Canberra Hospital/Lung Foundation Australia, Canberra/Brisbane, ACT, AUSTRALIA
Timely promotion was difficult.
Objectives: Objectives: Australia has one of the highest incidence rates of malignant mesothelioma in the world[1]. There
is increasing evidence that a third; non-occupational wave of
mesothelioma cases has developed in Australia, with a projected increase of 79% by 2020[2] . Thus given the vast nature of
the Australian continent, it was believed that it was necessary
to provide health professionals with current best practice guidelines and resources about asbestos related diseases and specifically malignant pleural mesothelioma (ARD & MPM). A national
tour was conducted and included workshops/ forums for health
professionals utilising a multidisciplinary team approach. [1] https://www.mesothelioma-australia.com/media/10743/
amr-data-report_final-for-publication2013-1-.pdf [2] Kao SC,
Reid G, Lee K, Vardy J, Clarke S, van Zandwijk N. Malignant
Mesothelioma. Internal Medicine Journal. [Review]. 2010;
40:742–50
Accessing rural/remote staff to attend workshops due to staffing levels was a major challenge and required workshops timing
to be altered.
Conclusion: As the Australian mesothelioma projected figures
increase over the next decade, we as a nation wish to be
prepared to provide personalised care to this cohort of patients
and carers. The Australian tour presented evidence based,
one day workshops to health professionals across the nation
to improve their understanding of diagnosis, treatments, legal
matters and how and where to access available resources for
ARD/MPM patients, carers and families to provide professional
“personalised care”.
Keywords: Mesothelioma, Australia, personalised care, Australian, health professional, tour, successful, Australian first,
education
Methods: To meet the personalised needs of this increasing
number of patients who are scattered extensively across the
nation, two Australian organisations collaborated to provide
workshops based on Best Practice Guidelines, current research
and clinical practice to health professionals. These one day
workshops were conducted in each capital city and one rural/
regional destination in each state and territory. There were
modified workshops for indigenous health professionals and
GP’s. In each destination suitable venues were identified,
advertising conducted and catering organised. Presenters with
clinical expertise were sought in each area from local respiratory physicians, medical and radiation oncologists, palliative
care specialist nurses, surgeons, allied health and the legal
fraternity. Specialists were flown in where local specialists were
not available. The program consisted of: introduction/history
of ARD/MPM; diagnosis, pathology, staging and treatment;
palliative care needs; the psychological impact and the legal
considerations for patients and their carers with MPM. All
sessions were evaluated by participants and all attendees were
awarded certificates of attendance.
Results: A total of 18 workshops/forums were conducted in
Australia for health professionals with an attendance of 551
participants. Eight were conducted in the capital cities, ten in
rural/regional There were modified workshops for indigenous
health professional and GP’s. The most valued features of the
workshops as identified by the participant’s evaluations were:
Quality of the workshop content and the relevance to daily
practice
Quality, knowledge and enthusiasm of the speakers
Multidisciplinary approach
Opportunity for professional networking
Quality of the written resources
Challenges for the project:
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AUTHOR INDEX
Authors highlighted in bold =
Presenting Author for this abstract
A
Abal, Mariana.....................................................................................PP01.33
Abate-Daga, Daniel............................................................................MS04.04
Abdel-Rahman, Abdel-Rahman M.......................PP01.35, PP01.42, PP01.53
Abdelrahman, Abdelrahman M..........................................................PP02.09
Aboelazm, Omnia............................................................................... PP01.61
Achard, Carole...................................................................................MS04.02
Acuña, Claudia.................................................................................... PP01.33
Aderca, Ileana....................................................................................MS04.06
Adusumilli, Prasad S.......................................................................... PL05.01
Aerts, Joachim.................................................. MS16.03, PP02.21, PP02.22,
.......................................................................................... PP02.33, PP02.36
Aeschlimann, Beat............................................................................. MS15.05
Agostini, Lorenzo...............................................................................MS05.05
Ahiskali, Rengin.................................................................................. PP02.47
Ahmed, Jaylan.................................................................................... PP01.61
Aigner, Clemens................................................................................. MS15.02
Ak, Guntulu....................................... MS06.04, PP01.25, PP02.47, PP02.53
Akın Kabalak, Pınar............................................................................ PP01.25
Al-Taei, Saly.......................................................................................MS16.05
Albelda, Steven............................................... MS04.05, MS04.06, MS16.01,
...........................................................................................PL06.05, PP01.28
Aldieri, Elisabetta............................................................................... PP01.72
Alexander, Laura................................................................................. PP01.02
Alfieri, Roberta................................................................................... MS13.02
Aliverti, Andrea....................................................................................PP01.14
Allcock, Richard J............................................................................... PP01.70
Allen, Louise........................................................................................ PP01.12
Alley, Evan.......................................................... MS03.02, PL06.05, PP02.08
Allione, Alessandra............................................................................. PP01.72
Almutairi, Abdulhadi......................................................................... PP02.29
Altomonte, Maresa............................................................................ MS10.03
Alves Das Mercês, Nen Nalú.............................................................. PP01.60
Alì, Greta............................................................................................MS08.04
Ambrogi, Marcello C...........................................................MS15.03, PP01.17
Ambrogi, Vincenzo............................................................................ PP02.46
Ambrosi, Jean-Paul............................................................................. PP01.50
Amodio, Rosalba................................................................................. PP01.59
Ampollini, Luca..................................................................................PP01.20
Ananthanarayanan, Viju....................................................................MS08.05
Anbunathan, Hima............................................................................. MS07.02
Ancona, Laura....................................................................MS08.06, PP01.44
Andersen, Claus B.............................................................................. PP01.69
Andersen, Morten............................................................................... PP01.69
Andreassen, Trygve............................................................................ PP01.21
Angelillo, Italo..................................................................................... PP01.59
Annesi, Diego..................................................................................... MS10.03
Antoine, Daniel J................................................................................MS05.03
Aoe, Keisuke........................................................................................PP02.04
Aou El-Kasem, Fatma M.A.............PP01.35, PP01.42, PP01.53, PP02.09
Aprile, Vittorio.................................................................................... MS15.03
Ardissone, Francesco......................................................................... PP01.56
Arif, Qudsia......................................................................................... PP01.22
Armato, Sam......................................................................................MS03.06
Armstrong, Bruce............................................................................... PP01.43
Arni, Stephan......................................................................................MS11.01
Arns, Madeleine.................................................................................. PP01.62
Asano, Michiko................................................................................... PP01.34
Ascoli, Valeria............................... MS07.03, MS08.06, PP01.59, PP01.64
Ashizawa, Kazuto................................................................................PP02.04
Ashton, Miranda J.............................................................................MS14.02
Aspesi, Anna....................................................................................... PP01.64
Aston, Wayne J..................................................... PP01.77, PP01.78, PP02.26
Atagi, Shinji........................................................................................ MS10.07
Atinkaya Ozturk, Cansel..................................................................... PP01.25
Avella Patino, Diego............................................................................ PL05.06
Aziz Shokralla, Hala........................................PP01.29, PP01.35, PP01.37,
..........................................................................PP01.38, PP01.39, PP01.53
B
Baas, Paul.......................................................... MS04.07, PP02.33, PP02.36
Bakó, Szilvia.......................................................................................MS02.05
Baldassarre, Antonio.......................................................................... PP01.44
Ball, Hannah......................................................................................PP02.76
Ballarino, Paola................................................................................. MS12.03
Baratti, Dario....................................................................................MS12.01
Barba, Adalberto................................................................................ MS10.02
Barbieri, Pietro G...............................................................................MS06.03
Barbiero, Fabiano..............................................................................MS06.02
Barbone, Dario................................................................... MS02.07, PP02.54
Barbone, Fabio...................................................................................MS06.02
Bareggi, Claudia................................................................................. PP01.07
Barlow, Julianne................................................................................ MS10.04
Barnes, Daniel T..................................................................................PP02.06
Barone, Elisa..................................................................................... PP02.63
Bates, Gleneara E...........................................MS12.02, MS15.07, PP02.02
Bathen, Tone F.................................................................................... PP01.21
Batirel, Hasan.................................................................. PP01.26, PP02.47
Battistini, Francesca........................................................................... PP01.59
Baumann, Francine...........................................MS05.03, MS07.04, PP01.50
Baumgartner, Bernhard...................................................................... PP01.62
Bearz, Alessandra............................................................................... PP01.31
Beatty, Gregoary................................................................................. PP01.28
Behner, Dusty.................................................................................... MS07.04
Bella, Francesca................................................................................. PP01.59
Belvedere, Ornella........................................................................... MS06.02
Bendell, Johanna C............................................................................ MS10.01
Benepal, Taqdir.................................................................................. MS07.02
Benfatto, Lucia.................................................................................... PP01.59
Bennett, Jonathan.......................................... MS02.02, MS02.06, MS11.03,
............................................................................ MS11.06, PL01.06, PP02.75
Benziane, Sarah.................................................................................PP01.11
Bérard, Karima...................................................................MS11.01, PP01.06
Berardi, Rosanna................................................................................PP02.33
Berger, Ian B.................................................................... MS03.02, PP02.08
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ABSTRACT BOOK
Berger, Walter..................................... PP02.60, PP02.61, PP02.64, PP02.69
Bertazzi, Pier Alberto.......................................................................... PP01.41
Bertocci, Erica................................................................................... MS10.03
Bertoglio, Pietro................................................................ MS15.03, PP01.17
Bertulli, Rossella................................................................................ MS12.01
Betti, Marta..........................................................PP01.56, PP01.64, PP01.72
Bezemer, Koen................................................... MS16.03, PP02.21, PP02.22
Biasi, Alessandra................................................................ PP01.56, PP01.64
Biesma, Bonne....................................................................................PP02.36
Biffo, Stefano..................................................................................... MS13.02
Bijou, Fontanet.................................................................................... PL04.03
Bilancia, Rocco..................................................................................MS09.04
Bilgi, Zeynep....................................................................................... PP02.47
Birembaut, Philippe............................................................ PP01.08, PP02.28
Bishop, Paul........................................................................................ PP01.68
Bitanihirwe, Byron K.Y........................................................................ PP01.06
Bittinger, Mark................................................................................... MS10.04
Blakemore, Stephen J........................................................................ PL04.03
Blanquart, Christophe.....................................MS02.03, PP01.09, PP01.11
Blazek, Gertrude................................................................................. PP01.62
Blum, Walter......................................................PP02.15, PP02.19, PP02.58
Blum, Yuna....................................................................................... MS08.08
Blumenschein, Jr, George R.............................................................. MS10.01
Blyth, Kevin.................................................. MS03.03, MS04.08, MS10.06,
.............................................................................................PP01.01, PP01.02
Boisgerault, Nicolas..........................................................................MS04.02
Boissard, Alice.................................................................................... PP01.08
Boldorini, Renzo................................................................. MS05.04, PP01.64
Bolitschek, Josef................................................................................. PP01.62
Bollati, Valentina................................................................................. PP01.07
Bölükbas, Servet............................................................................... PP02.44
Bolyard, Chelsea................................................................................MS04.08
Bomalaski, John................................................................................ MS10.02
Bommeli, Cordelia............................................................................. MS15.05
Bonotti, Alessandra...........................................................MS05.04, PP02.63
Booton, Richard................................................................. MS05.01, PP01.68
Bordini, Lorenzo.................................................................................. PP01.07
Borg, Elaine....................................................................................... MS13.04
Borroni, Ester..................................................................................... MS11.02
Boss, Andreas.....................................................................................MS11.01
Botta, Laura........................................................................................ PP01.59
Botticella, Angela..............................................................................MS03.04
Bowcock, Anne M.............................................................................. MS07.02
Boyer, Michael.................................................................. MS04.03, MS10.05
Bradbury, Penelope............................................................................ PL02.03
Braggio, Cesare.................................................................................. PP01.20
Brahmbhatt, Himanshu..................................................................... MS10.05
Braidwood, Lynne..............................................................................MS04.08
Brcic, Luka......................................................................... MS08.03, PP01.40
Brentisci, Carol................................................................................... PP01.59
Bressan, Vittoria..................................................PP01.30, PP01.44, PP01.59
Bressler, Yaakov............................................... MS12.02, MS15.07, PP02.02
Brina, Daniela.................................................................................... MS13.02
Broaddus, Courtney..........................................................MS02.07, PP02.54
Brochard, Patrick...............................................................................MS08.07
Broggini-Tenzer, Angela......................................................................MS11.01
Broome, Richard................................................................................MS06.05
Brozik, Jan..........................................................................................PP02.06
Bruno, Caterina................................................................................... PP01.46
Bruno, Rossella................................................................................ MS08.04
Bryant-Greenwood, Peter.................................................................. MS07.04
Buck, Brenda...................................................................................... PP01.50
Bueno, Raphael...............................................MS02.07, MS07.01, MS10.04,
............................................................................ PP01.06, PP02.54, PP02.71
Buikhuisen, Wieneke..........................................MS04.07, PP02.27, PP02.36
Burgers, Sjaak..................................... MS04.07, PP01.19, PP02.27, PP02.36
Burton, Kimberley.............................................................................. MS13.01
Busacca, Sara......................................................................PP01.71, PP01.73
Bushell, Martin.................................. MS02.02, MS11.06, PL01.06, PP02.75
Bydder, Sean.......................................................................................PP02.23
C
Cabras, Antonello D........................................................................... MS12.01
Caglar, Hale B..................................................................................... PP02.47
Cain, Kelvin...................................................... MS02.02, MS02.06, MS11.06
Calabro’, Luana.................................................................................MS10.03
Caldarella, Adele................................................................................ PP01.59
Cale, Alexander...................................................................................PP02.48
Campanini, Nicoletta.......................................................................... PP01.20
Candela, Pina...................................................................................... PP01.59
Canino, Claudia.................................................................................MS05.03
Capocaccia, Riccardo......................................................................... PP01.59
Carbognani, Paolo.............................................................................. PP01.20
Carbone, Michele..........................MS05.03, MS07.04, PP01.50, PP01.76
Carlson, Sephanie.............................................................................MS04.06
Carnovale Scalzo, Caterina...............................................................MS08.06
Casadio, Caterina............................................................... PP01.56, PP01.64
Casalone, Elisabetta.........................................PP01.56, PP01.64, PP01.72
Casbard, Angela.................................................................................PP02.34
Cascione, Luciano.............................................................................MS05.04
Case, Bruce W....................................................................................PP01.66
Casey, Thomas................................................................................... MS16.07
Cassetti, Paolo...................................................................................MS06.02
Cattaneo, Monica..............................................................................MS06.02
Cavaletto, Maria................................................................................ MS11.02
Cavalleri, Tommaso............................................................................ PP01.07
Cellerin, Laurent................................................................................. PP01.09
Cena, Tiziana.......................................................................PP01.31, PP01.44
Cengel, Keith A................................................MS03.02, MS14.01, PP02.08
Cerkl, Peter......................................................................................... PP01.62
Chalmers, Anthony............................................................MS14.02, MS14.03
Chang, Yu-Yin...................................................................................... PP01.47
Chaturvedi, Anshuman...................................................... MS05.06, PP01.68
Chaudhry, Ikram.................................................................................PP02.29
Chaudhry, Mubarak............................................................................PP02.48
Chauhan, Anoop................................................................................ MS05.01
Cheah, Hui Min...................................................................................MS11.07
Chee, Jonathan...................................................................MS16.02, PP02.14
Cheema, Ahsan...................................................................................PP02.29
Chella, Antonio.................................................................................. MS15.03
Chellini, Elisabetta.............................................................. PP01.44, PP01.59
Chen, Tianhui................................................................................... MS06.06
Cheng, Yuen Yee.............................................MS04.03, MS11.04, PP02.59,
...........................................................................PP02.68, PP02.72, PP02.73
Chernova, Tatyana.......................................... MS02.02, MS02.06, MS11.03,
...........................................................................MS11.06, PL01.06, PP02.75
Chirieac, Lucian R...............................................................................MS07.01
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ABSTRACT BOOK
Cho, Bc John....................................................................................... PL02.03
Christensen, Brock C..........................................................................PP02.57
Church, W B........................................................................................ PP02.72
Chéné, Anne-Laure..............................................................................PP01.11
Ciammella, Patrizia...........................................................................MS05.05
Cirilli, Claudia..................................................................................... PP01.59
Clarke, Stephen................................................................................. MS10.05
Clayson, Helen................................................ MS01.04, MS01.05, MS01.06
Cleaver, Amanda.................................................................................PP02.26
Clive, Amelia O................................................................................... MS14.05
Cocchioni, Mario................................................................................. PP01.59
Cognetti, Francesco........................................................................... PP01.31
Coindre, Jean-Michele....................................................................... PL04.03
Collins, Colin....................................................................................... PP01.65
Colombo, Enrico................................................................................. PP01.64
Comba, Pietro..................................................................................... PP01.46
Conner, Joe....................................................................... MS04.08, MS10.06
Consonni, Dario............................... PP01.07, PP01.41, PP01.59, PP01.60
Conti, Susanna..................................................................................PP01.46
Contiero, Paolo................................................................................... PP01.59
Cook, Alistair M................................................................................ PP02.23
Cookson, William O........................................................................... MS07.02
Coolen, Johan.................................................................................. MS03.04
Coonar, Aman..................................................................................... PP02.31
Cooper, Wendy A...............................................................................MS04.03
Coosemans, Willy..............................................................................MS03.04
Copin, Marie-Christine...................................................... MS08.08, PL02.05
Coqueret, Olivier................................................................................. PP01.08
Cornelissen, Robin..............................................................PP02.22, PP02.36
Corti, Mariangela................................................................................ PP01.59
Costa, Chrisostome...........................................................................MS02.04
Cowan, Katherine.............................................................................. MS01.05
Cowell, Gordon W..............................................................................MS03.03
Cowen, Michael..................................................................................PP02.48
Cozzi, Ilaria....................................................................... MS07.03, MS08.06
Creaney, Jenette................. MS11.07, MS16.02, PP01.23, PP01.70, PP02.14
Creech, Lorraine.............................................................................. MS05.06
Cristaudo, Alfonso.............................................................MS05.04, PP02.63
Croce, Carlo M................................................................................... MS13.02
Crosbie, Philip..................................................................................... PP01.68
Cuccaro, Francesco............................................................................ PP01.44
Cucevic, Branka.................................................................................. PP01.40
Cusimano, Rosanna............................................................................ PP01.59
Czirok, Andras.....................................................................................PP02.55
D
D’Angelo, Massimo.............................................................MS12.03, PP01.31
Daga, Antonio.................................................................................... MS11.02
Daimon, Takashi................................................................................ MS13.05
Dalcher, Damian.................................................................................PP02.57
Dallari, Barbara...................................................................PP01.41, PP01.59
Dammeijer, Floris..............................................................MS16.03, PP02.21
Danielli, Riccardo.............................................................................. MS10.03
Danson, Sarah................................................................................... MS10.06
Dao, Nhien.......................................................................................... PP02.71
Dao, Tao.............................................................................................. PL05.05
Dayal, Saurabh.................................................................................. MS14.03
De Angelis, Roberta............................................................................ PP01.59
De Fonseka, Duneesha......................................................................PP01.12
De Giorgi, Anna Maria........................................................................ PP01.59
De Klerk, Nicholas.............................................................................PP01.30
De Koning, Leanne............................................................................. PL04.05
De La Maza-Borja, Luis....................................................................MS14.04
De Leyn, Paul.................................................................... MS03.04, MS15.01
De Lima, Vinicius................................................................................ PP01.27
De Matteis, Sara................................................................................. PP01.41
De Perrot, Marc............................................... MS14.04, PL02.03, PP02.15,
.............................................................................PP02.18, PP02.19, PP02.74
De Reyniès, Aurélien..........................................................................MS08.08
De Rienzo, Assunta........................................................... MS07.01, PP02.71
De Santi, Chiara............................................................................... MS05.04
De Smet, Ruben.................................................................................MS05.02
De Waele, Jorrit.................................................................................. PP02.10
De Wever, Walter................................................................................MS03.04
Decaluwé, Herbert.............................................................................MS03.04
Degiovanni, Daniela...........................................................MS12.03, PP01.31
Dekeyser, Melanie............................................................................. MS15.01
Dekeyzer, Frederik.............................................................................MS03.04
Delanghe, Joris R...............................................................................MS05.02
Delanghe, Sigurd...............................................................................MS05.02
Delaunay, Tiphaine............................................................................MS04.02
Della Gatta, Andrew.......................................................................... MS11.04
Delvenne, Philippe.............................................................................MS02.04
Demery, Amr....................................................................................... PP01.53
Depypere, Lieven...............................................................................MS03.04
Deraco, Marcello............................................................................... MS12.01
Deshayes, Sophie...............................................................MS02.03, PP01.11
Devo, Perry........................................................................................PP02.11
Değirmenci, Irfan...............................................................................PP02.53
Dhalluin, Xavier...................................................................................PP02.37
Dhanasingh, Immanuel...................................................................... PP01.04
Dhont, Ludovic................................................................................. MS02.04
Di Gaetano, Cornelia...........................................................PP01.56, PP01.72
Di Giacomo, Anna Maria.................................................................... MS10.03
Di Pietrantonj, Carlo........................................................................... PP02.41
Dianzani, Caterina.............................................................................. PP01.64
Dianzani, Irma................................................... PP01.56, PP01.64, PP01.72
Diaz, Monica...................................................................................... MS10.02
Dick, Craig.........................................................................................MS03.03
Dick, Ian M.......................................................................................... PP01.70
Dieckmann, Karin.............................................................................. MS15.02
Dietz, Allen.........................................................................................MS04.06
Dini, Paolo.......................................................................................... MS15.03
Dinsdale, David..................................................MS02.06, MS11.06, PL01.06
Dioni, Laura........................................................................................ PP01.07
Djearaman, Madava............................................................................PP01.14
Dobbs, Adrian....................................................................................PP02.17
Dobra, Katalin.................................................... PP01.05, PP01.13, PP02.67
Dogan, A. Umran................................................................................ PP01.50
Dogan, Meral...................................................................................... PP01.50
Dome, Balazs...................................... MS15.02, PP01.15, PP01.17, PP01.18,
.............................................PP01.19, PP02.55, PP02.61, PP02.64, PP02.69
Donaldson, Ken...................................................................................PL01.06
Dong, Yawen......................... PP01.15, PP01.17, PP01.18, PP01.62, PP02.69
Dongel, Isa.......................................................................................... PP01.25
Dooms, Christophe........................................................... MS03.04, MS15.01
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ABSTRACT BOOK
Driscoll, Tim........................................................................................ PP01.43
Dubbeldam, Adriana..........................................................................MS03.04
Dudek, Kate....................................................................................... MS11.06
Duff, Anthony P................................................................................... PP02.72
Dujmovic, Tomislav............................................................................. PP01.40
Durkin, Amy........................................................................................ PL05.06
Duysinx, Bernard...............................................................................MS02.04
E
Eberlein, Michael................................................................................PP02.44
Ebert, Martin.......................................................................................PP02.23
Eckmayr, Josef.................................................................................... PP01.62
Edelman, J J........................................................................................PP02.59
Edwards, John...................................................MS05.01, MS10.06, PP02.31
El Bastawisy, Ahmed.........................................................................PP01.61
El Soud Youssef, Magdolen................................................................PP02.36
Elarouci, Nabila.................................................................................MS08.08
Eldemery, Amr M.............................................................................. PP02.09
Elshafie, Ghazi...................................................................................PP01.14
Emi, Mitsuru....................................................................................... MS07.04
Errhalt, Peter....................................................................................... PP01.62
Ersoz, Elcin......................................................................................... PP01.25
Evison, Matthew................................................................ MS05.06, PP01.68
Fisichella, Pietro M.............................................................................PP02.39
Flicker, Martin..................................................................................... PP01.62
Flores, Erin..........................................................................MS07.04, PP01.50
Foddis, Rudy......................................................................MS05.04, PP02.63
Follador, Alessandro..........................................................................MS06.02
Follo, Carlo...................................................................... MS02.07, PP02.54
Fontanini, Gabriella...........................................................................MS08.04
Fonteneau, Jean-François.................................................................MS04.02
Foot, Heather..................................................................................... MS01.05
Forastiere, Francesco........................................................ MS08.06, PP01.59
Formisano, Debora............................................................................MS05.05
Forte, Iris M......................................................................................... PP02.13
Foschi, Roberto................................................................................... PP01.59
Foster, Helen...................................................................................... MS10.05
Foster, John E.................................................................... MS03.03, PP01.02
Fox-Rushy, Julia.................................................................................. PP02.31
Francis, David..................................................................................... PP01.63
Francis, Roslyn....................................................................................PP02.23
Franklin, Peter..................................................................................... PP01.30
Frasca, Graziella................................................................................. PP01.59
Frauenfelder, Thomas.......................................................MS03.05, MS09.02
Friedberg, Joseph...............................................................................PP02.08
Friess, Martina................................................ MS03.05, MS09.02, MS15.05,
............................................................................................ PP01.06, PP01.24
Fuchimoto, Yasuko.............................................................................. PP01.34
Fujimoto, Nobukazu...........................................................PP01.34, PP02.04
Furukawa, Masashi.............................................................................PP02.05
Fusco, Mario....................................................................................... PP01.59
G
F
Facciolo, Francesco........................................................................... MS07.03
Falcini, Fabio....................................................................................... PP01.59
Falco, Francesco................................................................................MS05.05
Falzon, Mary...................................................................................... MS13.04
Fanetti, Anna Clara............................................................................. PP01.59
Fanucchi, Olivia................................................................................. MS15.03
Fasola, Gianpiero...............................................................................MS06.02
Favero, Chiara..................................................................................... PP01.07
Fazzo, Lucia........................................................................................ PP01.46
Feld, Ronald........................................................................................ PL02.03
Felley-Bosco, Emanuela.................................... MS11.01, PP01.06, PP02.15,
............................................................................PP02.19, PP02.56, PP02.57,
...........................................................................................PP02.58, PP02.70
Fennell, Dean....................................MS05.01, MS11.02, MS11.03, PP01.71,
............................................................PP01.73, PP01.74, PP02.33, PP02.34,
............................................................................................PP02.54, PP02.65
Feola, Daniela..................................................................................... PP01.59
Ferrante, Daniela................................PL01.04, PP01.44, PP01.56, PP01.64
Ferroni, Patrizia.................................................................................MS04.04
Filice, Angelina..................................................................................MS05.05
Findik, Gokturk................................................................................... PP01.25
Fisher, Patricia................................................................................... MS10.06
Fisher, Scott....................................................MS13.01, MS16.07, PP01.77,
...........................................................................PP01.78, PP02.23, PP02.26
Gaafar, Rabab M.................................................PP01.35, PP01.37, PP01.38,
.............................................................PP01.39, PP01.53, PP02.09, PP02.32
Galanis, Evanthia...............................................................................MS04.06
Galateau-Sallé, Françoise............... MS08.07, MS08.08, PL02.05, PP02.37
Galavotti, Sara...............................................MS02.02, MS02.06, MS11.03,
.............................................................................................PL01.06, PP02.75
Galeone, Carla...................................................................................MS05.05
Gallan, Alexander J............................................................................MS08.05
Gallizzi, Giulia.................................................................................... MS12.03
Gangemi, Manuela............................................................................. PP01.59
Gao, Zhi Bin........................................................................................MS08.02
Garassino, Marina Chiara................................................................... PP01.59
Garay, Tamas...................................................................................... PP02.61
Garcia Gerardi, Carlos........................................................................ PP01.33
Gardner, Georgina..............................................................................PP02.34
Gasparini, Pierluigi........................................................... MS05.04, MS13.02
Gatta, Gemma..................................................................................... PP01.59
Gaudino, Giovanni............................................................................. MS07.04
Geltner, Christian................................................................................ PP01.62
Gemba, Kenichi...................................................................................PP02.04
Gemignani, Federica..........................................................MS05.04, PP02.63
Gennaro, Valerio................................................................................. PP01.59
Gennatas, Spyridon..........................................................................MS07.02
Gercovich, Felipe Gustavo.................................................................. PP01.33
Ghaly, Galal......................................................................................... PP01.37
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ABSTRACT BOOK
Giangreco, Adam................................................................................ PL04.06
Giangreco, Giovanni...........................................................................PP02.63
Giangreco, Manuela..........................................................................MS06.02
Giannarelli, Diana.............................................................................. MS10.03
Giannini, Riccardo.............................................................................MS08.04
Giavarra, Marco.................................................................................MS06.02
Gil Deza, Ernesto................................................................................ PP01.33
Gilg Soit Ilg, Anabelle........................................................................MS08.07
Gill, Ritu............................................................................................. MS10.04
Gilmour, Lesley.................................................................................MS14.03
Ginsberg, Michelle...............................................................PL05.01, PL05.05
Giordano, Antonio............................................................................... PP02.13
Girardi, Paolo.....................................................................................MS06.03
Gironi, Laura C.................................................................................... PP01.64
Gittins, Jacki...................................................................................... MS05.01
Giuliani, Orietta................................................................................... PP01.59
Giurdanella, Maria Concetta.............................................................. PP01.59
Glodic, Goran...................................................................................... PP01.40
Gnetti, Letizia...................................................................................... PP01.20
Godfrey, Jack..................................................................................... MS11.06
Gola, Gemma...................................................................................... PP01.59
Goldoni, Matteo.................................................................................. PP01.20
Gomez, Daniel R................................................................................. PL05.01
Goparaju, Chandra M........................................................................MS05.03
Grauslund, Morten.............................................................................. PP01.69
Greaves, Peter......................................................................................PL01.06
Greillier, Laurent.................................................................................PP02.37
Grillo, Lucia Rosalba.......................................................................... MS07.03
Groen, Harry.......................................................................................PP02.36
Grosso, Federica..............................................MS12.03, PP01.31, PP01.64
Grosso, Stefano............................................. MS02.02, MS11.03, MS11.06,
............................................................................MS13.02, PL01.06, PP02.75
Großer, Christian.................................................................................PP02.43
Grusch, Michael................................................ PP01.19, PP02.60, PP02.61,
...........................................................................................PP02.64, PP02.69
Grutters, Jan.......................................................................................PP02.27
Grégoire, Marc................................................. MS02.03, MS04.02, PP01.08,
.............................................................PP01.09, PP01.11, PP02.28, PP02.62
Guadagni, Fiorella.............................................................................MS04.04
Guaglio, Marcello............................................................................... MS12.01
Guarrera, Simonetta............................................PP01.56, PP01.64, PP01.72
Guaschino, Roberto............................................................................ PP01.56
Guazzelli, Alice................................................................................... MS13.06
Gudmundsson, Eyjolfur................................................................... MS03.06
Guenther, Detlef................................................................................. MS15.05
Guette, Catherine............................................................................... PP01.08
Gueugnon, Fabien............................................................................... PP01.09
Guneş, Hasan Veysi............................................................................PP02.53
Guo, Holly........................................................................................... PP02.18
Gustafson, Mike.................................................................................MS04.06
H
Hagmolen Of Ten Have, Wanda....................................................... PP02.27
Håkansson, Anders P.......................................................................... PP02.72
Hamaidia, Malik.................................................................................PP02.12
Hamasaki, Makoto.............................................................................MS08.01
Harries, Lauren..................................................................................PP01.68
Hasegawa, Mizue..............................................................MS08.02, PP01.32
Hasegawa, Seiki............................................. MS09.06, MS13.05, PP02.07,
............................................................................ PP02.49, PP02.50, PP02.51
Hashimoto, Masaki............................ MS09.06, PP02.49, PP02.50, PP02.51
Hassan, Raffit............................... MS04.04, MS10.01, PP02.16, PP02.35
Hastings, Robert..................................................................PP01.71, PP01.73
Hatz, Rudolf A.................................................................................... MS15.04
Hayashi, Tatsurou...............................................................................PP02.05
Heffeter, Petra.....................................................................................PP02.69
Hegedus, Balazs..................................................PP01.15, PP01.17, PP01.18,
............................................................................PP01.19, PP02.55, PP02.61, ............................................................................................PP02.64, PP02.69
Hegmans, Joost................................. MS16.03, PP02.21, PP02.22, PP02.33
Heidari-Hamedani, Ghazal.................................................................PP02.67
Hemminki, Kari..................................................................................MS06.06
Hendriks, Rudi................................................................................... MS16.03
Hens, Niel........................................................................................... PP02.10
Hermans, Christophe......................................................................... PP02.10
Hesdorffer, Mary............................................................................... MS07.04
Hida, Tomoyuki..................................................................................MS08.01
Hilberg, Frank..................................................................................... PP02.61
Hill, Kate...........................................................................................MS01.05
Hill, Kate M........................................................................................ MS01.06
Hillerdal, Carl-Olof.............................................................................PP01.13
Hillerdal, Gunnar.............................................................. MS06.07, PP01.05
Hillinger, Sven...................................................................MS03.05, MS09.02
Hiroshima, Kenzo...........................................MS08.02, MS08.05, PP01.32,
............................................................................................PP02.52, PP02.66
Hjerpe, Anders................................................... PP01.05, PP01.13, PP02.67
Hmeljak, Julija..................................................................................MS13.03
Ho, Peter T.........................................................................................PL04.03
Hoda, Mir A......................................MS15.02, PP01.15, PP01.17, PP01.18,
.......................................................... PP01.19, PP01.62, PP02.60, PP02.61,
.......................................................................................... PP02.64, PP02.69
Hodgson, Clare................................................................................... PP01.68
Hoffman, Harriet................................................................................ MS07.04
Hoffman, Kimberly.............................................................................PP02.35
Hofman, Paul..................................................................... MS08.08, PL02.05
Hofmann, Hans-Stefan.......................................................................PP02.43
Holme, Jayne..................................................................... MS05.06, PP01.68
Hope, Danika E................................................................................... PP01.77
Hosteler, B.......................................................................................... PP01.23
Howell, Viive....................................................................................... PP02.72
Huang, Quin........................................................................................PP02.39
Hubert, Pascale.................................................................................MS02.04
Hudson, Amanda L............................................................................. PP02.72
Humphreys, Catherine A...................................................................MS03.03
Husain, Aliya.................................... MS08.05, MS16.06, PL05.06, PP01.22
Hussain, Michelle.............................................................................. MS13.06
Huynh, Yennie.................................................................................... MS10.05
Hveem, Kristian.................................................................................. PP01.21
Hylebos, Marieke...............................................................................PP01.67
Haas, Andrew.....................................................................MS03.02, PP02.08
Haas, Andrew R.................................................................................. PP01.28
Hagan, Sarah..................................................................................... MS14.01
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I
Iannuzzi, Carmelina A.......................................................................PP02.13
Imbeaud, Sandrine............................................................................. PL02.05
Inai, Kouki...........................................................................................PP02.04
Inal Cengiz, Tuba................................................................................ PP01.25
Inci, Ilhan..........................................................................MS03.05, MS09.02
Indovina, Paola................................................................................... PP02.13
Ingles-Prieto, Alvaro...........................................................................PP02.60
Isai, Dona Greta..................................................................................PP02.55
Ishibashi, Hironori............................................................. MS15.06, PP02.45
Italiano, Antoine.................................................................................. PL04.03
Iwasaki, Ainori...................................................................................MS08.01
J
Jackson, Andrew................................................................................PL05.02
Jackson, Sue......................................................................................MS05.06
Jakopovic, Marko...............................................................................PP01.40
Janes, Sam........................................................................ MS13.04, PL04.06
Janevski, Zoran................................................................................... PP01.40
Janovjak, Harald.................................................................................PP02.60
Jarrold, Ian......................................................................................... MS01.05
Jaurand, Marie Claude...................................... MS08.08, PL02.05, PL04.05
Jean, Didier........................................................ MS08.08, PL02.05, PL04.05
Jennings, Cormac............................................................................... PP01.50
Jetter, Alexander................................................................................ MS15.05
Jitesh, Puthen V..................................................................................PP02.54
Jithesh, Puthen V............................................................... MS11.02, PP02.65
Johnson, Kevin C................................................................................PP02.57
Johnson, Thomas...............................................................................PP02.73
Johnson, Todd.................................................................................... MS07.04
Johnston, Amanda............................................................................. MS10.02
Jones, Carolyn................................................................................... MS11.06
Jørnild, Alina....................................................................................... PP01.69
June, Carl............................................................................................ PP01.28
K
Kadota, Yoshihisa.............................................................................. MS10.07
Kaijen-Lambers, Margaretha............................................ MS16.03, PP02.22
Kainrath, Stephanie............................................................................PP02.60
Kamata, Toshiko................................................................................. PP01.32
Kamel, Mohamed............................................................................... PP01.39
Kamikonya, Norihiko......................................................... MS09.06, PP02.51
Kaneda, Yasufumi.............................................................................. MS10.07
Kanteti, Rajani..................................................................................... PP01.22
Kanteti, Rajani P.................................................................................PP01.04
Kao, Steven....................................................... MS04.03, MS10.05, PP02.56
Kapaklikaya, Esra...............................................................................PP02.57
Kasa, Alma......................................................................................... MS12.03
Kato, Katsuya.................................................................................... PP02.04
Katou, Katsuya....................................................................................PP02.05
Katsura, Hideki................................................................................... PP01.32
Katz, Sharyn.......................................................................................PP02.08
Katz, Sharyn I.................................................................................. MS03.02
Kaur, Balveen.....................................................................................MS04.08
Kawahara, Kunimitsu........................................................................MS08.01
Kazan, Steven.................................................................. PP01.36, PP01.75
Keating, Jane...................................................................MS03.01, MS04.05
Keilhack, Heike................................................................................... PL04.03
Kelly, Caroline..................................................................................... PP01.02
Kenkel, David......................................................................................MS11.01
Kern, Izidor.........................................................................................MS08.03
Kerr, Naseem...................................................................................... PP01.28
Khadeir, Ramsay................................................................................ MS10.02
Khalid, Urooj......................................................................MS03.02, PP02.08
Khalil, Mohammed........................................................................... PP02.48
Khanna, Swati.................................................................... MS04.04, PP02.16
Kharazmi, Elham...............................................................................MS06.06
Khattri, Arun...................................................................................... MS16.06
Khong, Andrea....................................................................................PP02.26
Kindler, Hedy...................................................MS03.06, MS10.01, MS16.06,
.............................................................................PL05.06, PP01.04, PP01.22
Kirchbacher, Klaus............................................................................. PP01.62
Kircheva, Diana................................................................................... PL05.06
Kirschner, Michaela........................................................................... MS07.04
Kirschner, Michaela B.................................... MS04.03, MS15.05, PP01.19,
...........................................................PP01.24, PP02.56, PP02.59, PP02.68
Kirwan, Marie....................................................................................MS05.06
Kishimoto, Takumi.............................................PP01.34, PP02.03, PP02.04
Kızılgöz, Derya.................................................................................... PP01.25
Klabatsa, Astero................................................................................PL06.05
Klebe, Sonja....................................................... MS04.03, PP02.59, PP02.73
Klepetko, Walter..................................................MS15.02, PP01.15, PP01.17,
............................................................................ PP01.18, PP01.19, PP01.62, ............................................................................................ PP02.61, PP02.69
Klikovits, Thomas.............................MS15.02, PP01.15, PP01.17, PP01.18,
............................................................. PP01.19, PP01.62, PP02.61, PP02.69
Klotz, Laura V....................................................................................MS15.04
Knight, Julia........................................................................................PP02.63
Knott, Robert B................................................................................... PP02.72
Kobayashi, Masashi..........................................................MS15.06, PP02.45
Kodnar, Julia....................................................................................... PP01.15
Koh, Eitetsu......................................................................................... PP01.32
Koller, Francisco José......................................................................... PP01.60
Kolluri, Krishna.................................................................MS13.04, PL04.06
Kondo, Nobuyuki............................ MS09.06, PP02.49, PP02.50, PP02.51
Korasidis, Stylianos........................................................................... MS15.03
Korse, Catharina M............................................................................. PP01.19
Kossenkov, Andrew............................................................................. PL06.05
Kratzke, Robert..................................................................................MS04.06
Krausz, Thomas.................................................................................MS08.05
Kresoja-Rakic, Jelena......................................................PP02.56, PP02.57
Krowczynska, Małgorzata...................................................PP01.51, PP01.54
Krstic-Demonacos, Marija................................................................MS13.06
Krug, Lee..............................................................................PL05.01, PL05.05
Kryeziu, Kushtrim...............................................................................PP02.69
Kucharczuk, John.............................................................................. MS03.01
Kudo, Tomoo...................................................................................... MS13.05
Kukulj, Suzana.................................................................................... PP01.40
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ABSTRACT BOOK
Kumar, Neelam.................................................................MS13.04, PL04.06
Kumar, Prem........................................................................................PP01.14
Kumazawa, Sachiko.......................................................... MS15.06, PP02.45
Kuo, Licheng....................................................................................... PL05.02
Kuribayashi, Kozo............................................. PP01.03, PP01.52, PP02.07,
............................................................................................ PP02.49, PP02.51
Kuroda, Ayumi................................... MS09.06, PP02.49, PP02.50, PP02.51
Kusamura, Shigeki............................................................................. MS12.01
L
Labby, Zacariah E..............................................................................MS03.06
Lacey, Simon...................................................................................... PP01.28
Lacin, Tunc.......................................................................................... PP02.47
Ladanyi, Marc.................................................................................... MS13.03
Lagani, Vincenzo................................................................................. PP01.21
Lagniau, Sabrina................................................................................PP01.16
Laird, Barry........................................................................................ MS14.02
Lakatos, Dora...................................................................................... PP02.61
Lake, Richard.....................................................MS13.01, MS16.07, PP01.77,
.............................................................PP01.78, PP01.79, PP02.23, PP02.26
Lam, Vincent..................................................................................... PP02.06
Lamote, Kevin................................................................... MS05.02, PP01.16
Landi, Stefano....................................................................MS05.04, PP02.63
Lando, Cecilia Francesca................................................................... PP01.59
Lang, Georg....................................................................................... MS15.02
Lansley, Sally M................................................ MS11.07, MS16.07, PP02.23
Lantuejoul, Sylvie..............................................................................MS08.07
Larson, David...................................................................................... PP01.50
Larus, John......................................................................................... PL04.03
Laszlo, Viktoria...................MS15.02, PP01.15, PP01.17, PP02.61, PP02.69
Lathrop, Mark.................................................................................... MS07.02
Lauk, Olivia...................................... MS07.04, MS09.02, MS11.01, MS15.05
Le Pimpec-Barthes, Françoise...........MS08.08, PL02.05, PL04.05, PP02.37
Le Quesne, John................................MS11.03, MS11.06, PL01.06, PP01.63,
..............................................................................PP01.71, PP01.73, PP01.74
Le Stang, Nolwenn............................................................................MS08.07
Learmonth, Kirsty..............................................................................MS04.08
Lebenthal, Abraham......................................................................... PP02.39
Lee, Chunman...................................................................................MS10.07
Lee, Lukas J.......................................................................................PP01.47
Lee, Yc Gary........................................................................................MS11.07
Legittimo, Patrizia............................................................................... PP01.44
Leigh, James....................................................................................... PP01.43
Leighl, Natasha................................................................................... PL02.03
Lennon, Frances E.............................................................................. PP01.04
Leslie, Felicity.................................................................................... MS10.05
Lester, Jason F....................................................................................PP02.34
Lesterhuis, Willem J........................................ MS13.01, MS16.07, PP01.77,
..........................................................PP01.78, PP01.79, PP02.23, PP02.26
Li, Feng...............................................................................................MS03.06
Libener, Roberta................................................................. PP01.56, PP01.64
Liddell, Charly..................................................................................... PP01.09
Lievens, Yolande................................................................................ MS15.01
Lievense, Lysanne........................................... MS16.03, PP02.21, PP02.22
Lim, Eric............................................................................MS07.02, PP02.30
Lin, Ruby............................................................................ MS11.04, PP02.64
Lin, Ruby C.......................................................... PP02.59, PP02.68, PP02.73
Lind, Michael J................................................................................... MS11.05
Lindner, Michael................................................................................ MS15.04
Linton, Anthony...............................................MS04.03, MS06.05, MS10.05
Litzky, Leslie....................................................................................... PL06.05
Liu, Guo J............................................................................................ PP02.72
Lizotte, Patrick................................................................................... MS10.04
Lo, Albert............................................................................................ MS16.01
Loay, Eman.........................................................................................PP02.09
Loay, Iman........................................................................................... PP01.38
Locatelli, Myriam................................................................................PP02.37
Lococo, Filippo..................................................................................MS05.05
Loembe, Arsene-Bienvenu.................................................................PP02.32
Lou, Jianlin.........................................................................................MS06.06
Loubani, Mahmoud.............................................................................PP02.48
Louis, Renaud....................................................................................MS02.04
Lowe, Val............................................................................................MS04.06
Lu, Shir Kiong.................................................................................... MS07.02
Luberto, Ferdinando........................................................................... PP01.44
Lucchi, Marco.................................................. MS05.04, MS08.04, MS15.03
Lum, Trina..........................................................................................MS04.03
M
Ma, Shaokang.....................................................................MS16.02, PP02.14
Maceyko, Steve.................................................................................. MS16.01
Macfarlane, Marion........................................ MS02.02, MS02.06, MS11.03,
............................................................................ MS11.06, PL01.06, PP02.75
Machan, Barbara................................................................................ PP01.62
Macleod, Nicholas............................................................................. MS14.02
Madore, Jason...................................................................................MS04.03
Maffè, Antonella................................................................................. PP01.64
Magnani, Corrado..............................................PL01.04, PP01.31, PP01.44,
............................................................PP01.56, PP01.64, PP01.72, PP02.41
Maher, Stephen G.............................................................................. MS11.05
Maio, Michele.................................................................................... MS10.03
Malcervelli, Gabriela.......................................................................... PP01.33
Maldonado, Fabien............................................................................MS04.06
Mallone, Sandra................................................................................. PP01.59
Mandrekar, Sumithra.........................................................................MS04.06
Manente, Arcangela G....................................................... MS11.02, PP02.65
Mangone, Lucia.................................................................................. PP01.59
Manno, Valerio.................................................................................... PP01.46
Mansano Sarquis, Leila Maria........................................................... PP01.60
Mansfield, Aaron S...........................................................MS04.06, PP01.10
Mantovani, Maria De Fátima.............................................................. PP01.60
Marchevsky, Alberto..........................................................................MS08.05
Marciniak, Stefan J............................................................ MS05.01, PP02.63
Marcq, Elly.........................................................................................PP02.10
Margery, Jacques...............................................................................PP02.37
Marinaccio, Alessandro...................................................................... PP01.44
Markaki, Maria.................................................................................... PP01.21
Martinson, Luke.................................................................................PP01.74
Mascaux, Céline................................................................................MS02.04
Maskell, Nick....................................MS05.01, MS14.05, PP01.12, PP02.31
Massard, Christophe.......................................................................... PL04.03
Matsuda, Tsuyoshi............................................................................. MS01.04
Matsumoto, Seiji................................................ MS09.06, PP02.49, PP02.51
Matsumoto, Shinji..............................................................................MS08.01
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ABSTRACT BOOK
Mattioli, Stefano................................................................................. PP01.44
Mattone, Silvia.................................................................................... PP02.41
Matullo, Giuseppe................................................PP01.56, PP01.64, PP01.72
Maza, Luis D.L.................................................................................... PP02.18
Mazuranic, Anton................................................................................ PP01.40
Mazuranic, Ivica................................................................................. PP01.40
Mccaughan, Brian C.......................................... MS04.03, PP02.59, PP02.73
Mcclanahan, Terri............................................................................... PL06.05
Mcdiarmid, Jennifer........................................................................... MS10.05
Mcdonald, Alice.................................................................................. PL04.03
Mcdonnell, Alison...............................................................................PP02.23
Mcgarvey, Maureen............................................................................ PP01.28
Mcgettrick, Michael...........................................................................PP01.01
Mcgregor, Stephanie.........................................................................MS08.05
Mclaughlin, Megan.............................................................................PP02.35
Mclaurin, Brett.................................................................................... PP01.50
Mclean, Jocelyn................................................................................ PP02.42
Meduri, Stefano.................................................................................MS06.02
Meerang, Mayura........................... MS11.01, MS15.05, PP01.06, PP01.24
Mehes, Elod........................................................................................PP02.55
Meiller, Clément.................................................................................. PL02.05
Melaiu, Ombretta..............................................................MS05.04, PP02.63
Melenhorst, Jos.................................................................................. PP01.28
Melfi, Franca......................................................................................MS08.04
Menegozzo, Simona........................................................................... PP01.44
Mensi, Carolina.....................................PP01.07, PP01.41, PP01.59, PP01.60
Meristoudis, Christos......................................................................... MS07.05
Merkler, Doug..................................................................................... PP01.50
Merler, Enzo.....................................................................MS06.03, PP01.30,
............................................................................................ PP01.44, PP01.59
Mesopath, Experts Pathologists.......................................................MS08.07
Metintas, Muzaffer.........................................MS06.01, MS06.04, PP01.25,
.............................................................................PP02.47, PP02.53, PP02.67
Metintas, Selma................................................MS06.04, PP01.25, PP02.53
Miccoli, Sara....................................................................................... PP01.64
Michiara, Maria................................................................................... PP01.59
Michot, Jean-Marie............................................................................. PL04.03
Middleton, Gary.................................................................................. PP02.16
Mikami, Koji.......................................................PP01.03, PP01.52, PP02.07
Miles, Gareth J................................................. MS02.02, MS02.06, MS11.06
Miligi, Lucia......................................................................................... PP01.44
Millward, Michael J............................................................................. PP01.79
Miluzio, Annarita................................................................................ MS13.02
Minaai, Michael................................................................................. MS07.04
Minelli, Giada..................................................................... MS08.06, PP01.46
Mineo, Tommaso Claudio...................................................................PP02.46
Minkiewicz, Anna............................................................. PP01.51, PP01.54
Mirabelli, Dario...................................................PL01.04, PP01.44, PP01.56,
.............................................................................................PP01.59, PP01.64
Miyamoto, Yosuke............................................................................... PP01.34
Mjelle, Robin....................................................................................... PP01.21
Moffatt, Miriam.................................................................................. MS07.02
Mohamed, Abdelrahman.....................................PP01.37, PP01.38, PP01.39
Molina, Julian....................................................................................MS04.06
Montero, Angeles.............................................................. MS07.02, MS08.05
Moody, Hannah L.............................................................................MS11.05
Moon, Edmund................................. MS14.01, MS16.01, PL06.05, PP01.28
Moons, Johnny................................................................................... MS15.01
Moore, David A................................... PP01.63, PP01.71, PP01.73, PP01.74
Moore, Kathleen N............................................................................ MS10.01
Mora, Marco......................................................................................MS05.04
Morahan, Grant................................................................................. MS13.01
Morgenfeld, Eduardo.......................................................................... PP01.33
Morinaga, Takao.................................................................PP02.52, PP02.66
Moro, Laura.....................................................MS02.05, MS11.02, PP02.65
Morra, Aldo........................................................................................ MS10.03
Morre, Dj............................................................................................. PP01.23
Morre, Dm........................................................................................... PP01.23
Morris, Paul......................................................................................... PP01.50
Morrow, Betsy.................................................................... MS04.04, PP02.16
Moseley, Jennifer................................................................................PP02.39
Moser, Justin........................................................................................PP01.10
Motas, Natalia.................................................................................... PP02.41
Munck, Camille....................................................................................PP01.11
Murer, Bruno..................................................................... MS05.04, MS13.02
Murphy, Erin....................................................................................... PL06.05
Murphy, Fiona A...................................................................PL01.06, PP02.75
Musk, Arthur W................................................................... PP01.23, PP01.30
Mussai, Francis................................................................................... PP02.16
Mussi, Alfredo....................................................MS08.04, MS15.03, PP01.17
Musti, Marina...................................................................................... PP01.44
Mutti, Antonio..................................................................................... PP01.20
Mutti, Luciano.................................. MS05.04, MS13.02, MS13.06, PP02.63
Muzio, Alberto.....................................................................MS12.03, PP01.31
N
Nabavi, Noushin................................................................................PP01.65
Nabeshima, Kazuki..........................................................MS08.01, MS08.05
Nackaerts, Kristiaan......................................................... MS03.04, MS15.01
Nader, Joëlle................................... MS04.02, PP01.08, PP02.28, PP02.62
Nafteux, Philippe.............................................................MS03.04, MS15.01
Nagamatsu, Yasuko S.......................................................................MS01.04
Naidu, Babu.........................................................................................PP01.14
Nakamichi, Toru................................MS09.06, PP02.49, PP02.50, PP02.51
Nakano, Takashi.............................. MS09.06, MS10.07, MS13.05, PP01.03,
............................................................. PP01.52, PP02.07, PP02.49, PP02.51
Nakas, Apostolos............................................ MS02.02, MS02.06, MS09.01,
......................................................... MS09.04, MS11.03, MS11.06, PL01.06,
..............................................................PP01.71, PP01.73, PP01.74, PP02.75
Napolitano, Andrea............................................MS05.03, MS07.04, PP01.50
Nasu, Masaki..................................................................................... MS07.04
Nawrocki-Raby, Béatrice.................................................... PP01.08, PP02.28
Neelature Sriramareddy, Sathya......................................................MS02.04
Nelson, Annemarie............................................................................. PP01.28
Nelson, Gill........................................................................................PP01.57
Nemunaitis, John............................................................................... MS10.01
Neu, Reiner.........................................................................................PP02.43
Neufeld, Zoltan...................................................................................PP02.55
Newick, Kheng................................................................................... MS16.01
Nguyen, Jean-Michel.......................................................................... PP01.09
Nguyen-Kim, Thi Dan Linh...............................................MS03.05, MS09.02
Nicholson, Andrew............................................................................. MS07.02
Nicolson, Marianne............................................................................ PP02.41
Niewiadomsky, Dario.......................................................................... PP01.33
Nilsson, Stefan.................................................................. MS11.02, PP02.65
Nims, Sarah...................................................................... MS03.01, MS04.05
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ABSTRACT BOOK
Nishi, Hideyuki.................................................................................... PP01.34
Nixon, Lisette S................................................................................. PP02.34
Nowak, Anna.........................PP01.77, PP01.78, PP01.79, PP02.23, PP02.32
Numico, Gianmauro............................................................MS12.03, PP01.31
O
O’Brien, Shaun................................................................................... MS16.01
O’Brien, Mary E.................................................................................. MS07.02
O’Hara, Mark...................................................................................... PP01.28
O’Rourke, Noelle................................................................................ MS14.02
Oda, Yoshinao....................................................................................MS08.01
Oddone, Enrico................................................................................... PP01.44
Oehler, Rudolf..................................................................................... PP01.15
Ogliara, Paola..................................................................................... PP01.64
Ojo, Anthonia...................................................................................... PL05.02
Okabayashi, Asako............................................................................. PP01.32
Okabe, Kazunori............................................................................... PP02.05
Okamoto, Shinya................................................................................PP02.66
Okubo, Kenichi..................................................................MS15.06, PP02.45
Okumura, Meinoshin......................................................................... MS10.07
Okumura, Yoshitomo......................................................... MS09.06, PP02.51
Oledzka, Gabriela................................................................PP01.51, PP01.54
Oliveto, Stefania...............................................................................MS13.02
Olschweski, Horst............................................................................... PP01.62
Oneglia, Barbara................................................................................ MS12.03
Op De Beeck, Ken............................................................................... PP01.67
Opitz, Isabelle.............................................. MS03.05, MS07.04, MS09.02,
.........................................................................MS11.01, MS15.05, PP01.06, ............................................................................. PP01.19, PP01.24, PP02.70
Orlandi, Augusto................................................................................MS04.04
Otsuki, Taiichiro..................................PP01.03, PP01.52, PP02.07, PP02.51
Ottolini, Barbara..................................................................................PP01.74
Ötvöss, Rita......................................................................................... PP01.13
Own, Sulaf A....................................................................................... PP01.05
Ozaki, Daisuke...................................................................................MS08.02
Ozaki, Shinji........................................................................................ PP01.34
Ozsvar, Judit....................................................................................... PP02.61
Ozturk, Akın........................................................................................ PP01.25
P
Pacey, Simon..................................................................................... MS10.02
Pachter, Jonathan...............................................................................PP02.21
Paci, Massimiliano.............................................................................MS05.05
Pagano, Ian........................................................................................MS05.03
Pairon, Jean Claude...........................................................................MS08.07
Paku, Sandor......................................................................................PP02.55
Palleschi, Alessandro......................................................................... PP01.07
Panageas, Katherine.......................................................................... PL05.05
Panou, Vasiliki...................................................................................MS07.05
Papotti, Mauro....................................................................MS07.03, PP01.64
Pardini, Barbara.................................................................................. PP01.72
Parhar, Preeti...................................................................................... PL05.01
Pascucci, Cristiana............................................................................. PP01.59
Pasini, Barbara................................................................................... PP01.64
Pass, Harvey I....................................................MS05.03, MS07.04, PP01.50
Passaro, Carmela............................................................................... PP02.13
Pastan, Ira........................................................................................... PP02.16
Pastorino, Sandra..............................................MS05.03, MS07.04, PP01.50
Patel, Akash.......................................................................MS03.02, PP02.08
Patel, Manish.....................................................................................MS04.06
Pattison, Scott................................................................................... MS10.05
Paul, Jim............................................................................................. PP01.02
Pauwels, Patrick................................................................................. PP02.10
Pavesi, Luca........................................................................................PP02.65
Pavlakis, Nick..................................................................................... MS10.05
Pavone, Venere................................................................................... PP01.44
Paz-Ares, Luis..................................................................................... PP02.41
Pearson, John..................................................................................... PP01.70
Pecze, Laszlo......................................................................................PP02.58
Pedrazzoli, Paolo................................................................................ PP01.31
Peek, Ann............................................................................................ PP01.02
Peeters, Stephanie........................................................... MS03.04, MS15.01
Peikert, Tobias..................................................................MS04.06, PP01.10
Pellegrini, Laura................................................................. MS05.03, PP01.50
Pellizzari, Giacomo............................................................................MS06.02
Pennati, Marzia.................................................................................. MS12.01
Pentimalli, Francesca......................................................................... PP02.13
Pernazza, Fausto............................................................................... MS12.03
Perrone, Federica.............................................................................. MS12.01
Perticaroli, Patrizia............................................................................. PP01.44
Pesatori, Angela C.............................................................................. PP01.07
Pesce, Elisa........................................................................................ MS13.02
Petel, Fabien......................................................................................MS08.08
Peters, Susan...................................................................................... PP01.30
Petrucci, Maria Saba.......................................................................... PP01.59
Pettinari, Aldo..................................................................................... PP01.44
Phillips, Melissa................................................................................. MS10.02
Piccolini, Ezio..................................................................... MS12.03, PP01.56
Piffer, Silvano...................................................................................... PP01.59
Pinto, Carmine.................................................................................... PP01.31
Pinton, Giulia................................................. MS02.05, MS11.02, PP02.65
Pirastu, Roberta.................................................................................. PP01.44
Pirker, Christine..................................................................................PP02.69
Piro, Roberto...................................................................... MS05.05, PP01.59
Pisa, Frederica...................................................................................MS06.02
Pissaloux, Daniel...............................................................................MS08.07
Planchard, David.................................................................PP02.37, PP02.41
Plesa, Gabriela.................................................................................... PP01.28
Plestina, Sanja.................................................................................... PP01.40
Pohl, Wolfgang.................................................................................... PP01.62
Poland, Craig A....................................................................................PL01.06
Popat, Sanjay..................................................................... MS07.02, PP02.32
Popper, Helmut................................................................................... PP01.62
Portella, Giuseppe.............................................................................. PP02.13
Postel-Vinay, Sophie........................................................................... PL04.03
Pouliquen, Daniel L........................ MS04.02, PP01.08, PP02.28, PP02.62
Powers, Amy.......................................................................MS07.04, PP01.50
Powley, Ian R..................................................MS02.02, MS02.06, MS11.03,
.............................................................................................PL01.06, PP02.75
Predina, Jarrod................................................................MS03.01, MS04.05
Pringle, J. H........................................................................................ MS11.03
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ABSTRACT BOOK
Prokrym, Kirill....................................................................................MS05.03
Proto, Claudia..................................................................................... PP01.59
Puchalski Kalinke, Luciana................................................................ PP01.60
Q
Qadri, Syed.........................................................................................PP02.48
Quetel, Lisa........................................................................ PL02.05, PL04.05
Quirke, Phillip......................................................................PP01.71, PP01.73
Quispel-Janssen, Josine................................................. MS04.07, PP02.36
R
Rafferty, Judy.....................................................................................PP02.77
Rahman, Naj....................................................................................... PP02.31
Rahoma, Mohamed.............................................PP01.35, PP01.42, PP01.53
Rahouma, M..........................................PP01.37, PP01.38, PP01.39, PP01.61
Rahouma, Mohamed.......................................................................... PP01.29
Rajagopalan, Prabhu......................................................................... MS10.01
Ranucci, Alessandra........................................................................... PP01.44
Rapicetta, Cristian........................................................................... MS05.05
Rassl, Doris M.................................................................... MS05.01, PP02.63
Rath, Emma M.................................................................................... PP02.72
Ravn, Jesper........................................................................................ PP01.69
Rea, Federico...................................................................................... PP01.31
Reck, Martin........................................................................................PP02.32
Reichhart, Eva.....................................................................................PP02.60
Reid, Alison......................................................................................... PP01.30
Reid, Glen........................................MS04.03, MS10.05, MS11.04, PP01.19,
........................................................... PP01.24, PP02.56, PP02.59, PP02.64,
...........................................................................PP02.68, PP02.72, PP02.73
Renier, Annie...................................................... MS08.08, PL02.05, PL04.05
Ribich, Scott........................................................................................ PL04.03
Riboldi, Luciano.................................................................................. PP01.41
Ribrag, Vincent................................................................................... PL04.03
Ricciardi, Sara.................................................................................... MS13.02
Rice, David.......................................................................................... PL05.01
Richards, Cathy................................................................................... PP01.63
Richards, William................................................................MS10.04, PP01.06
Richards, William G........................... MS02.07, MS07.01, PP02.54, PP02.71
Ried, Michael.................................................................................... PP02.43
Riehm, Jacob J.................................................................................... PP01.04
Righi, Luisella.....................................................................MS07.03, PP01.64
Rimanti, Anita..................................................................................... PP01.59
Rimner, Andreas................................................................ PL05.01, PL05.02
Rinaldi, Catherine A.............................................MS11.07, PP01.78, PP01.79
Rintoul, Robert C............................................................ MS05.01, PP02.31
Rizzello, Roberto Vito......................................................................... PP01.59
Robard, Myriam.................................................MS04.02, PP02.28, PP02.62
Robinson, Bruce............................MS13.01, MS16.02, MS16.07, PP01.23,
........................................................... PP01.70, PP02.14, PP02.23, PP02.26
Robinson, Cleo.................................................................................... PP01.50
Roche, Maria....................................................................................... PL04.03
Roden, Anja........................................................................ MS04.06, PP01.10
Røe, Oluf D........................................................................ MS07.05, PP01.21
Roglic, Mihovil.................................................................................... PP01.40
Rolli, Luigi........................................................................................... PP01.20
Romanelli, Antonio............................................................................. PP01.59
Romeo, Elisa...................................................... MS08.06, PP01.44, PP01.59
Rooijackers, Jos..................................................................................PP02.27
Rosato, Simonetta.............................................................................. PP01.64
Rose, Buerkley................................................................... MS03.06, PL05.06
Rosenzweig, Kenneth E...................................................................... PL05.01
Rosolen, Valentina.............................................................................MS06.02
Roulois, David....................................................................................MS02.03
Roveta, Annalisa.................................................................MS12.03, PP01.31
Ruffini, Enrico..................................................................................... PP01.56
Ruiz, William F..................................................................................... PP01.36
Rullo, Emma.......................................................................................MS08.06
Rusca, Michele................................................................................... PP01.20
Rusch, Valerie......................................................................PL05.01, PL05.05
Rush, Hannah.................................................................................... MS10.02
S
Sagan, Christine................................................................................. PP01.09
Sage, Beth........................................................................................... PL04.06
Sage, Elizabeth.................................................................................. MS13.04
Saito, Atsuhiro................................................................................... MS10.07
Sala, Orietta........................................................................................ PP01.44
Salgia, Ravi......................................................................... PP01.04, PP01.22
Salimu, Josephine............................................................................. MS16.05
Samarzija, Miroslav............................................................................ PP01.40
Santin, Alessandro D......................................................................... MS10.01
Santoni-Rugiu, Eric............................................................................PP01.69
Santoro, Armando.............................................................................. PP01.31
Santoro, Raffaella...............................................................................PP02.57
Sarapa, Nenad................................................................................... MS10.01
Sarun, Kadir........................................................................ PP02.68, PP02.72
Sarun, Kadir H....................................................................................PP02.59
Sato, Akitoshi...................................................................................... PP01.32
Sato, Ayuko....................................................................... MS08.01, MS13.05
Sblattero, Daniele..............................................................................MS02.05
Scagliotti, Giorgio V........................................... PP01.31, PP02.32, PP02.41
Scarnato, Corrado.............................................................................. PP01.44
Scheinberg, David.............................................................................. PL05.05
Schelch, Karin.....................................................PP02.60, PP02.64, PP02.69
Schenk, Peter...................................................................................... PP01.62
Scherpereel, Arnaud.........................................MS02.04, MS08.07, PP01.11,
.......................................................................................... PP02.33, PP02.37
Schinwald, Anja...................................................................................PL01.06
Schlom, Jeffrey..................................................................................MS04.04
Schmalzl, Angelika.............................................................................PP02.67
Schmid-Bindert, Gerald...................................................................... PP02.41
Schmidt, Anna.................................................................................... PL04.03
Schneiter, Didier................................................................................MS09.02
Schwaller, Beat...................................................PP02.15, PP02.19, PP02.56,
............................................................................................ PP02.57, PP02.58
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ABSTRACT BOOK
Schwartz, Jean-Marc......................................................................... MS13.06
Sciacca, Salvatore.............................................................................. PP01.59
Sciaranghella, Daniele....................................................................... PP02.71
Scolyer, Richard A..............................................................................MS04.03
Scuderi, Tiziana.................................................................................. PP01.59
Seifert, Burkhardt............................................. MS03.05, MS09.02, PP01.06
Seiwert, Tanguy Y.............................................................................. MS16.06
Seiwerth, Sven.................................................................................... PP01.40
Sekido, Yoshitaka...............................................................................PL01.09
Sekine, Ikuo........................................................................................PP02.66
Sekine, Yasuo..................................................................... MS08.02, PP01.32
Seward, Shelly M............................................................................... MS10.01
Sgarbi, Giorgio...................................................................................MS05.05
Sgargi, Paolo....................................................................................... PP01.59
Shah, Rajesh...................................................................... MS05.06, PP01.68
Sharkey, Annabel J...................................... MS09.01, MS09.04, PP01.71,
........................................................................................... PP01.73, PP01.74
Sharples, Linda................................................................................... PP02.31
Shaw, Jacqui........................................................................................PP01.74
Sheaff, Michael.................................................................................. MS10.02
Shen, Ronglai...................................................................................... PL05.01
Sherlock-Brown, Graham................................................................... PP02.31
Shibata, Eisuke................................................................... PP01.03, PP02.07
Shimada, Hideaki..............................................MS08.02, PP02.52, PP02.66
Shimizu, Shigeki................................................................................ MS13.05
Shingyoji, Masato...............................................................................PP02.66
Shinohara, Yoshiyasu........................................................................ MS13.05
Shrestha, Raunak............................................................................... PP01.65
Sideris, Antonios................................................................................PP02.71
Siegert, Charles J...............................................................................PP02.39
Signorelli, Diego................................................................................. PP01.59
Silini, Enrico Maria............................................................................. PP01.20
Silvestri, Stefano................................................................................. PP01.44
Simone, Charles B.............................................MS03.02, MS14.01, PP02.08
Singhal, Sunil................................................... MS03.01, MS03.02, MS04.05
Siozopoulou, Vasiliki........................................................................... PP02.10
Sivasothy, Pasupathy.......................................................................... PP02.31
Sklarek, Jürgen.................................................................................. MS15.04
Skubiszewska, Agnieszka....................................................PP01.51, PP01.54
Smeele, Patrick....................................................................................PP01.11
Smit, Egbert........................................................................................ PP02.41
Smits, Evelien..................................................................................... PP02.10
Smojver-Jezek, Silvana....................................................................... PP01.40
Sneddon, Sophie...............................................................................PP01.70
Soeberg, Matthew J..........................................................MS06.05, PP01.43
Solin, Jessica......................................................................MS16.07, PP02.26
Solinas, Michela................................................................................. PP01.20
Soltermann, Alex................................................................................ PP01.06
Soluri, Maria Felicia...........................................................................MS02.05
Somigliana, Anna B...........................................................................MS06.03
Soria, Jean-Charles............................................................................ PL04.03
Sørensen, Jens B............................................................... PP01.27, PP01.69
Spicer, James..................................................................................... MS10.02
Spraggon, Lee.................................................................................... MS13.03
Spriggs, Ruth..................................................................................... MS11.06
Staal-Vd Brekel, Jeske........................................................................PP02.36
Stahel, Rolf......................................................MS09.02, MS11.01, MS15.05,
.............................................................PP01.06, PP02.56, PP02.57, PP02.70
Staumont, Bernard............................................................................MS02.04
Steele, Jeremy................................................................................... MS10.02
Steinberg, Seth..................................................................................MS04.04
Stella, Franco......................................................................................PP02.46
Stephens, Richard............................................................................. MS01.05
Sterman, Daniel................................................................. MS04.05, PL06.05
Stetler-Stevenson, Maryalice............................................................. PP02.16
Stobo, David B...................................................................................MS03.03
Stockhammer, Paul.............................. PP01.15, PP01.17, PP01.18, PP01.62
Storchi, Cinzia..................................................................................... PP01.59
Stracci, Fabrizio.................................................................................. PP01.59
Straus, Christopher M.......................................................................MS03.06
Studnicka, Michael............................................................................. PP01.62
Stura, Antonella.................................................................................. PP01.59
Suh, Hyerim....................................................................................... MS11.04
Sulemani, Merve.................................................................................PP02.56
Sun, Jing............................................................................................ MS16.01
Sun, Xiao-Ming..............................................MS02.02, MS02.06, MS11.03,
............................................................................ MS11.06, PL01.06, PP02.75
Sundquist, Kristina............................................................................MS06.06
Suzuki, Toshio.....................................................................PP02.52, PP02.66
Szatmari, Tände.................................................................................. PP01.13
Szatmari, Tünde..................................................................................PP02.67
Szeto, Kyle.......................................................................................... PP01.22
Szlosarek, Peter............................................................... MS05.01, MS10.02
Szöke, Tamas......................................................................................PP02.43
Sætrom, Pål........................................................................................ PP01.21
T
Tabi, Zsuzsanna............................................................... MS16.04, MS16.05
Tada, Yuji.......................................................... MS08.02, PP02.52, PP02.66
Taddei, Sofia......................................................................................MS05.05
Tagawa, Masatoshi.......................................... MS08.02, PP02.52, PP02.66
Taggart, Dj........................................................................................... PP01.23
Taghavi, Shahrokh............................................................................. MS15.02
Tagliabue, Giovanna........................................................................... PP01.59
Takasaki, Chihiro............................................................... MS15.06, PP02.45
Takeshima, Yukio................................................................................PP02.04
Takuwa, Teruhisa............................... MS09.06, PP02.49, PP02.50, PP02.51
Tan, Angelina.....................................................................................MS04.06
Tan, Yi-Hung Carol.......................................................... MS16.06, PP01.22
Tanaka, Fumihiro............................................................... MS09.06, PP02.51
Tanaka, Toshiki...................................................................................PP02.05
Tangy, Frédéric...................................................................................MS04.02
Tanji, Mika........................................................................................... PP01.50
Tanyi, Janos L..................................................................................... PP01.28
Tao, Hiroyuki.......................................................................................PP02.05
Tarnoki-Zach, Julia........................................................................... PP02.55
Tatsumi, Koichiro................................................................PP02.52, PP02.66
Taub, Robert N...................................................MS12.02, MS15.07, PP02.02
Taverna, Giacomo.............................................................................. MS12.03
Tavian, Daniela...................................................................................PP02.65
Taylor, Morag.......................................................................PP01.71, PP01.73
Taylor, Paul......................................................................... MS05.06, PP01.68
Telford, Nick........................................................................................ PP01.68
Tenconi, Sara................................................... MS05.05, MS09.01, MS09.04
Terada, Takayuki..................................................PP01.03, PP01.52, PP02.07
Terracini, Benedetto........................................................................... PL01.04
Tewaternaude, Jim.............................................................................. PP01.57
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ABSTRACT BOOK
Thomas, Anish................................................................... MS04.04, PP02.16
Thomson, Blythe................................................................................. PL04.03
Thurm, Rivka.......................................................................................PP02.02
Tinwell, Brendan................................................................................ MS07.02
Tironi, Andrea....................................................................................MS05.04
Tiseo, Marcello................................................................... PP01.20, PP01.59
Tod, Angela......................................................................................... PP02.31
Todaro, Aldo........................................................................................ PP01.07
Todisco, Liana.................................................................................... MS12.03
Tognelli, Flavio.................................................................................... PP01.33
Torigian, Drew A................................................................. MS03.02, PP01.28
Toulmonde, Maud............................................................................... PL04.03
Trama, Annalisa.................................................................................. PP01.59
Tranchant, Robin................................................................ PL02.05, PL04.05
Truini, Anna........................................................................................MS05.04
Tsamardinos, Ioannis.......................................................................... PP01.21
Tsao, Anne............................................................................PL05.01, PL05.05
Tsao, Ming........................................................................................... PP02.74
Tsim, Selina......................................................MS03.03, PP01.01, PP01.02
Tsubota, Noriaki................................................. MS09.06, PP02.49, PP02.51
Tsujimura, Tohru............................................MS08.01, MS09.06, MS13.05,
.............................................................................PP02.07, PP02.49, PP02.51
Tsunoda, Tatsuhiko............................................................................ MS07.04
Tumino, Rosario.................................................................................. PP01.59
Tuncel, Tunç...................................................................................... PP02.53
Tunesi, Sara.........................................................................PL01.04, PP01.56
Turchetti, Daniela............................................................................... PP01.64
V
Vaickus, Lou....................................................................................... MS10.04
Vaja, Ricky......................................................................... MS09.01, MS09.04
Valentino, Francesco.......................................................................... PP01.31
Vallely, Michael P................................................................................PP02.59
Van Audenaerde, Jonas...................................................................... PP02.10
Van Camp, Guy................................................................................... PP01.67
Van Dam, Loes....................................................................................PP02.54
Van Den Broek, Daan......................................................................... PP01.19
Van Heemst, Robbert.........................................................................PP02.36
Van Keulen, Virginia...........................................................................MS04.06
Van Meerbeeck, Jan.......................... MS05.02, PP02.10, PP02.32, PP02.33
Van Meerbeeck, Jan P.........................................................PP01.16, PP01.67
Van Nimwegen, Menno...................................................... MS16.03, PP02.21
Van Raemdonck, Dirk........................................................................MS03.04
Van Veer, Hans...................................................................................MS03.04
Van Zandwijk, Nico........................................MS04.03, MS06.05, MS10.05,
............................................................MS11.04, PP01.19, PP01.24, PP01.43,
............................................................ PP02.42, PP02.59, PP02.68, PP02.73
Vandecaveye, Vincent........................................................................MS03.04
Vandermeers, Fabian.........................................................................MS02.04
Vansteenkiste, Johan........................................................ MS03.04, MS15.01
Vatrano, Simona................................................................................ MS07.03
Vattiato, Rosa...................................................................................... PP01.59
Vd Noort, Vincent................................................................................PP02.36
Velema, Derek.....................................................................................PP02.32
Venegas, Ollin................................................................... MS03.01, MS04.05
Ventura, Luigi...................................................................................... PP01.20
Verbeken, Eric....................................................................................MS03.04
Verheij, Marcel................................................................................... MS14.03
Vermaelen, Karim Y.............................................................................PP01.16
Verschakelen, Johny..........................................................................MS03.04
Viberti, Clara........................................................................PP01.56, PP01.72
Vicentini, Massimo............................................................................. PP01.59
Vigneswaran, Wickii...........................................................................MS08.05
Vigneswaran, Wickii T........................................................PL05.06, PP01.22
Villaflor, Christopher........................................................................... PP01.22
Villamizar, Guillermo A......................................................................PP02.01
Visca, Paolo....................................................................................... MS07.03
Vitale, Maria Francesca...................................................................... PP01.59
Vlacic, Gregor.................................................................................. MS08.03
Von Wangenheim, Ute........................................................................PP02.32
Vrugt, Bart........................................................... MS11.01, PP01.06, PP01.24
Vyberg, Mogens................................................................................. MS07.05
W
Wada, Sae........................................................................................... PP01.34
Waddell, Nicola................................................................................... PP01.70
Wagner, Christina...............................................................................PP02.64
Wahba, Hisham................................................................... PP01.53, PP02.09
Waithman, Jason................................................................................PP02.23
Walker, Graeme................................................................................. MS13.01
Waller, David.....................................................MS09.01, MS09.04, PP01.71,
............................................................................. PP01.73, PP01.74, PP02.31
Wallin, Bruce A................................................................................. PP02.35
Walter, Annette.................................................................................. MS10.01
Walts, Ann..........................................................................................MS08.05
Wang, Jung-Der.................................................................................. PP01.47
Wang, Yan.......................................................................... MS10.04, PP02.21
Wang, Yuzhuo..................................................................................... PP01.65
Wasielewski, Eric.................................................................PP01.11, PP02.37
Wason, James.....................................................................................PP02.63
Watson, Sydeaka................................................................................ PL05.06
Weaver, David T.................................................................................. MS10.04
Weder, Walter.................................. MS03.05, MS07.04, MS09.02, MS11.01,
........................................................... MS15.05, PP01.06, PP01.19, PP01.24,
.............................................................................PP02.56, PP02.57, PP02.70
Weir, Chris........................................................................................... PP02.72
Whiting, Caroline............................................................................... MS01.05
Wilczynska, Ania................................................................................ MS11.06
Wild, Peter........................................................................................... PP01.24
Wilk, Ewa..............................................................................PP01.51, PP01.54
Willemin, Marie C................................................................................PP01.11
Willems, Luc....................................................................... MS02.04, PP02.12
Williams, Marissa........................................... MS04.03, MS11.04, PP01.19,
............................................................................................ PP02.59, PP02.73
Willis, Anne E.................................................. MS02.02, MS02.06, MS11.03,
............................................................................ MS11.06, PL01.06, PP02.75
Woll, Penella..................................................................... MS04.08, MS10.06
Won, Brian M...................................................................................... PP01.22
Wong, Kwok....................................................................................... MS10.04
Woo, Kaitlin M..................................................................................... PL05.01
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ABSTRACT BOOK
Woodward, Rosemary........................................................ MS03.03, PP01.02
Wright, Casey M..................................................................................PP02.59
Wu, Abraham J.................................................................................... PL05.01
Wu, Di................................................................................. MS08.02, PP01.32
Wu, Licun......................................................... MS14.04, PP02.15, PP02.18,
........................................................................................... PP02.19, PP02.74
Wu, Matthew.....................................................................MS14.04, PP02.18
Wu, Ting-Hui....................................................................................... PP01.47
Wuilleret, Guillaume.......................................................................... MS15.05
Zhou, Zhi............................................................................................ MS13.04
Ziino, Antonio...................................................................................... PP01.59
Ziltener, Gabriela.................................................................PP02.57, PP02.70
Zimmerman, Marion..........................................................................MS04.07
Zöchbauer-Müller, Sabine................................................................. MS15.02
Zonca, Sara....................................................................... MS02.05, MS11.02
Zucchi, Luigi.......................................................................................MS05.05
Zucman-Rossi, Jessica...................................... MS08.08, PL02.05, PL04.05
Zuo, Zhixiang...................................................................................... MS16.06
Zwaenepoel, Karen............................................................................. PP02.10
Y
Yahia, Maha........................................................................ PP01.61, PP02.09
Yamagishi, Tomoko............................................................................. PP01.34
Yan, Ming............................................................................................ PL05.02
Yang, Haining................................................... MS05.03, MS07.04, PP01.50
Yeap, Beow Y.......................................................................................MS07.01
Yearly, Jennifer H................................................................................ PL06.05
Yehia, Maha........................................................................................ PP01.53
Yildirim, Huseyin................................................................................. PP01.25
Yildizeli, Bedrettin............................................................................... PP02.47
Yilmaz, Senay..................................................................................... PP01.25
Yilmaz, Ulku........................................................................................ PP01.25
Yokohori, Naoko.................................................................................. PP01.32
Yorke, Ellen..........................................................................PL05.01, PL05.02
Yoshida, Kumiko.................................................................................PP02.05
Yoshiyama, Kouichi.............................................................................PP02.05
Young, Jane......................................................................................... PP01.43
Yuan, Constance................................................................................. PP02.16
Yuan, Zhenqiang................................................................................. PL04.06
Yuksel, Mustafa.................................................................................. PP02.47
Yumuk, Fulden.................................................................................... PP02.47
Yun, Hana............................................................................................ PP02.18
Yun, Hana Z......................................................................................... PP02.74
Yun, Zhihong.......................................................MS14.04, PP02.15, PP02.19
Yusa, Toshikazu..................................................................................MS08.02
Z
Zacarias-Cabeza, Joaquin................................................MS11.03, PP02.75
Zaffaroni, Nadia................................................................................. MS12.01
Zago, Giulia........................................................................................MS04.07
Zai, Silvia............................................................................................ MS12.03
Zanelli, Francesca.............................................................................MS05.05
Zanin, Tina.........................................................................................MS06.02
Zauderer, Marjorie G.......................................................... PL05.01, PL05.05
Zemek, Rachael M.............................................................................. PP01.79
Zhang, Jingli....................................................................... MS04.04, PP02.16
Zhang, Shu Dong................................................................................PP02.54
Zhang, Xing........................................................................................MS06.06
Zhao, Yidan D.....................................................................................PP02.74
iMig2016.ORG
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