Italian Association for the Study of the Liver – Annual Meeting

Transcription

Volume 47 Supplement 1 15 February 2015
ISSN 1590-8658
Digestive
and Liver Disease
An International Journal of Gastroenterology and Hepatology
Abstracts of the 48th A.I.S.F. – Italian Association for the Study of the
Liver – Annual Meeting 2015
Rome, 19-20 February 2015
Digestive
and Liver Disease
An International Journal of Gastroenterology and Hepatology
Vol. 47 Supplement 1 (2015)
Official Journal of:
Italian Society of Gastroenterology
French Society of Gastroenterology
Italian Association for the Study of the Liver
Italian Society for Digestive Endoscopy
Italian Association for the Study of the Pancreas
Italian Society for Pediatric Gastroenterology, Hepatology and Nutrition
Italian Association for Hospital Gastroenterologists and Digestive Endoscopists
Italian Group for the Study of Inflammatory Bowel Disease
Editor Emeritus
Gabriele Bianchi-Porro, Milan, Italy
Editorial Assistant
Serena Rotunno, Rome, Italy
Managing Editor
Paola Piccolo, Rome, Italy
Editor in Chief
Mario Angelico, Rome, Italy
Statistical Consultant
Alessandra Nardi, Rome, Italy
SECTION EDITORS
Inflammatory Bowel Disease
Laurent Peyrin-Biroulet, Vandoeuvre, France
Alessandro Armuzzi, Rome, Italy
General Gastroenterology
Elisabetta Buscarini, Crema, Italy
Cesare Hassan, Rome, Italy
Digestive Oncology
Thomas Aparicio, Bobigny, France
Côme Lepage, Dijon, France
Tamara Matysiak-Budnik, Nantes, France
Hepatology
Jérôme Dumortier, Lyon, France
Massimo Puoti, Milan, Italy
Sébastien Dharancy, Lille, France
Stefano Ginanni Corradini, Rome, Italy
Pediatric Gastroenterology
Salvatore Cucchiara, Rome, Italy
Clinical Methodology
Agostino Colli, Lecco, Italy
Digestive Endoscopy
Massimo Raimondo, Jacksonville, USA
Pier Alberto Testoni, Milan, Italy
Alberto Larghi, Rome, Italy
Pancreatic Disease
Massimo Falconi, Ancona, Italy
Gabriele Capurso, Rome, Italy
Basic Science
Gianfranco Alpini, Temple, USA
Romina Mancinelli, Rome, Italy
EDITORIAL BOARD
Domenico Alvaro, Rome, Italy
Angelo Andriulli, Foggia, Italy
Paolo Angeli, Padua, Italy
Adolfo Attili, Rome, Italy
Gabrio Bassotti, Perugia, Italy
Laurent Beaugerie, Paris, France
Robert Benamouzig, Bobigny, France
Antonio Benedetti, Ancona, Italy
Marc Benninga, Amsterdam, Netherlands
Marina Berenguer, Valencia, Spain
Roman Bogorad, Cambridge, USA
Jaime Bosch, Barcelona, Spain
Jean-Pierre Bronowicki, Vandoeuvre-Lès-Nancy, France
William Brugge, Boston, USA
Nicola Caporaso, Naples, Italy
Carlo Catassi, Ancona, Italy
Umberto Cillo, Padua, Italy
Dario Conte, Milan, Italy
Gino Roberto Corazza, Pavia, Italy
Enrico Corazziari, Rome, Italy
Antonio Craxì, Palermo, Italy
Roberto De Franchis, Milan, Italy
Gianfranco Delle Fave, Rome, Italy
Anthony Demetris, Pittsburgh, USA
Sharon DeMorrow, Temple, USA
Philippe Ducrotte, Rouen, France
Amal Dutta, Dallas, USA
Luca Elli, Milan, Italy
Stefano Fagiuoli, Bergamo, Italy
Luigi Familiari, Messina, Italy
Massimo Fantini, Rome, Italy
Piero M. Fisichella, Boston, USA
Heather Francis, Temple, USA
Mirella Fraquelli, Milan, Italy
Dennis Freshwater, Birmingham, UK
Giovanni B. Gaeta, Naples, Italy
Antonio Gasbarrini, Rome, Italy
Eugenio Gaudio, Rome, Italy
Shannon Glaser, Temple, USA
Pietro Invernizzi, Milan, Italy
Robert Jensen, Baltimore, USA
Michel Kahaleh, Charlottesville, USA
David Laharie, Pessac, France
René Laugier, Marseille, France
Astrid Lièvre, Saint-Cloud, France
Giovanni Maconi, Milan, Italy
Riccardo Marmo, Salerno, Italy
Marco Marzioni, Ancona, Italy
David Mutimer, Birmingham, UK
Mattijs Numans, Leiden, Netherlands
Jean-Marc Phelip, Saint Etienne, France
Antonio Pinna, Bologna, Italy
Franco Radaelli, Como, Italy
Alessandro Repici, Milan, Italy
Oliviero Riggio, Rome, Italy
Mario Rizzetto, Turin, Italy
Renato Romagnoli, Turin, Italy
Massimo Rugge, Padua, Italy
Tilman Sauerbruch, Bonn, Germany
Jean-Christophe Saurin, Pierre-Benite, France
Vincenzo Savarino, Genoa, Italy
Laurent Siproudhis, Rennes, France
Etienne Sokal, Brussels, Belgium
Mario Strazzabosco, New Haven, USA
Giacomo Carlo Sturniolo, Padua, Italy
Giuseppe Tisone, Rome, Italy
Michael Trauner, Vienna, Austria
Vincenzo Villanacci, Brescia, Italy
Frank Zerbib, Bordeaux, France
Contents
Vol. 47 Supplement 1 (15 February 2015)
Index Medicus (MEDLINE), Current Contents/Clinical Practice,
Science Citation Index and EMBASE/Excerpta Medica
Sociedad Iberoamericana de Información Cientı́fica (SIIC)
Associato alla Unione Stampa Periodica Italiana
Abstracts of the 48th A.I.S.F. – Italian Association
for the Study of the Liver – Annual Meeting 2015
Rome, 19–20 February 2015
Abstracts of the 48th A.I.S.F. – Italian Association for the Study of the Liver – Annual Meeting 2015:
Oral communications
e1
Poster sessions – THURSDAY
e19
Poster sessions – FRIDAY
e43
A.I.S.F. 2015: Abstracts evaluation procedure
e67
Digestive and Liver Disease 47S (2015) e1–e18
Contents lists available at ScienceDirect
Digestive and Liver Disease
journal homepage: www.elsevier.com/locate/dld
Liver – Annual Meeting 2015: Oral communications
OC-01
SELECTIVE LXR␣ INTESTINAL ACTIVATION
REDUCES HEPATIC INFLAMMATION AND
FIBROSIS DURING THE DEVELOPMENT OF
CHRONIC LIVER INJURY
I. Pierantonelli 1 , C. Pinto 1 , E. Mingarelli 1 ,
S. Saccomanno 1 , L. Agostinelli 1 , C. Rychlicki 1 ,
L. Trozzi 1 , A. Benedetti 1 , M. Marzioni 1 ,
A. Moschetta 2 , G. Svegliati-Baroni 1
1 Dipartimento di Gastroenterologia, Università
Politecnica delle Marche, Ancona, Italy
2 Dipartimento di Medicina Interdisciplinare,
Università di Bari, Bari, Italy
Background: Hepatic fibrosis represents the wound-healing
response of the liver to chronic injury characterized by increased
and altered deposition of newly generated extracellular matrix. The
pathogenesis of liver fibrosis is not fully understood, leading to a
lack in effective therapies. Liver X receptors (LXR ␣/␤) are important regulators of lipid metabolism. While in the liver LXRs regulate
cholesterol and fatty acid metabolism, intestinal LXR␣ activation
has been implicated in reverse cholesterol transport (RCT) pathway
leading to high levels of the anti-inflammatory HDL.
Aims: To evaluate the effect of the selective intestinal LXR␣ activation on the development of hepatic fibrosis associated to chronic
liver injury.
Methods: Male FVB/N mice with intestinal constitutive LXR␣
activation (iVP16LXR␣) and their control (iVP16) were treated with
twice weekly i.p. injection of Carbon Tetrachloride (1 ␮l/g body
weight) for 2 months.
Results: Immunohistochemistry for the macrophage marker
F4-80 indicated lowered infiltration in iVP16LXR␣ liver (p < 0.05).
Gene expression of the pro-inflammatory cytokines IL-6, TNF␣,
MCP-1 and NF-Kb were reduced in iVP16LXR␣-CCL4 mice (p < 0.05,
p < 0.05, p < 0.005 and p < 0.005 respectively). Collagen synthesis
and deposition was reduced in iVP16LXR␣ mice compared to iVP16
as determined by type-I-collagen and TGF␤ mRNA expression (both
p < 0.005) and Sirius Red morphometry (p < 0.0005). RCT induced by
intestinal LXR␣ activation was assessed by measuring plasmatic
HDL concentration and found increased in iVP16LXR␣-CCL4 mice
(p < 0.005). Such activation is not associated to hepatic de novo
1590-8658/$36.00
lipogenesis as shown by gene expression of SREBP1c and FAS and
decreased triglyceride content (p < 0.005).
Conclusions: Specific intestinal LXR␣ activation reduces liver
injury by increasing the level of the anti-inflammatory HDL cholesterol, thus leading to decreased hepatic fibrosis. Selective intestinal
activation of LXR␣ might be considered as a therapeutic approach
to reduce liver fibrosis avoiding the occurrence of hepatic steatosis
as a side effects associated to systemic LXR induction.
http://dx.doi.org/10.1016/j.dld.2015.01.007
OC-02
GUANFACINE (SPECIFIC ALPHA-2A
ADRENOCEPTOR AGONIST) RESTORES
NATRIURESIS IN EXPERIMENTAL CIRRHOTIC
ASCITES RESISTANT TO CLONIDINE AND
STANDARD DIURETICS
G. Sansoè 1 , M. Aragno 2 , R. Mastrocola 2 ,
M. Parola 2
1
Division of Gastroenterology, Gradenigo Hospital,
Torino, Turin, Italy
2 Department of Clinical and Biological Sciences,
University of Torino, Turin, Italy
Introduction: In human cirrhosis, adrenergic hyperfunction
causes proximal tubular fluid retention and contributes to refractory ascites; clonidine, a sympatholytic drug, plus diuretics
improves natriuresis in otherwise refractory ascites.
Aim: To compare clonidine (aspecific ␣2 -adrenoceptor agonist)
and SSP-002021R (specific ␣2A -receptor agonist and prodrug of
guanfacine), both associated with diuretics, in cirrhotic refractory
ascites.
Methods: Seven groups of 12 rats were studied: controls (G1);
controls receiving furosemide and potassium canrenoate (G2); rats
with ascitic cirrhosis due to 14 CCl4 weeks (G3); cirrhotic rats
treated with furosemide and canrenoate over the 11th–14th CCl4
weeks (G4); cirrhotic rats treated with furosemide, canrenoate
and clonidine (0.5 mcg) (G5); cirrhotic rats treated with diuretics
and low-dose clonidine (0.3 mcg) (G6); cirrhotic rats treated with
diuretics and SSP002021R (5 mg/kg b.w.) (G7). Three rats in each
group had their hormonal status and renal function assessed at the
end of 11th, 12th, 13th, and 14th CCl4 weeks.
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Abstracts / Digestive and Liver Disease 47S (2015) e1–e18
Results: Cirrhotic rats in G3 and G4 gained weight over the
11th–14th CCl4 weeks. In G4, brief increase in sodium excretion
due to diuretics (12th week) preceded worsening of inulin clearance and natriuresis (diuretic resistance). The addition of low-dose
clonidine (G6) or guanfacine (G7) to diuretics increased, respectively, electrolytes excretion over the 11th–12th CCl4 weeks, or
glomerular filtration rate (GFR) and electrolytes excretion over the
13th–14th CCl4 weeks. Natriuretic responses in G6 and G7 were
accompanied by significantly reduced catecholamines serum levels
vs. G3 and G4 (all P < 0.03).
Conclusions: Clonidine briefly potentiates diuretics-dependent
natriuresis. Conversely, guanfacine (␣2A -receptor agonist) restores
glomerular filtration rate and natriuresis, and prevents refractory
ascites in experimental advanced cirrhosis.
OC-03
THE PROTO-ONCOGENE TYROSINE-PROTEIN
KINASE MER AS A NOVEL MODULATOR OF
FIBROGENESIS IN NAFLD
G. Di Maira 1 , S. Petta 2 , A. Cappon 1 , E. Vivoli 1 ,
V. Di Marco 2 , F. Marra 1
1 Dipartimento di Medicina Sperimentale e Clinica,
University of Florence, Italy
2 Dipartimento Biomedico di Medicina Interna e
Specialistica, University of Palermo, Italy
Background and aims: MER (MERTK) is a receptor tyrosine
kinase of the TAM family often overexpressed or activated in
various malignancies. MERTK variants have been associated with
fibrosis severity at genome wide level in chronic hepatitis C, in
particular the variant rs4374383 G>A of MERTK gene has been associated with reduced liver fibrosis progression. However, currently
there is no available experimental evidence on the involvement of
MERTK in the modulation of the biology of hepatic stellate cells
(HSC) or in the fibrogenic process.
Methods: Primary human HSC were isolated from human livers and cultured on plastic. C57BL6/J mice were treated with CCl4
for 6 weeks. Balb/C mice were fed with a methionine and cholinedeficient (MCD) diet for 8 weeks. Intrahepatic gene expression was
assayed by quantitative real time PCR.
Results: Using immunoprecipitation, we found the expression
of MERTK in human HSC. Stimulation of human HSC with the
MERTK ligand GAS6 resulted in a time-dependent activation of
ERK1/2. Exposure to GAS6 also induced an enhancement of cell
migration of human HSC comparable to the effects of 10% FBS. In
both models of fibrosis (chronic CCl4 and MCD diet) we observed
a significant increase of expression of MERTK compared to control
group. Finally we studied the MERTK level in liver specimens of
patients with NASH with different degree of liver fibrosis. MERTK
expression was significantly higher in the group of patients with
more severe fibrosis.
Conclusions: In vitro and in vivo data indicate that MERTK could
play an important role of in the biology of HSC and the fibrogenic
process.
OC-04
THE CYSTIC FIBROSIS CONDUCTANCE REGULAR
(CFTR) CONTROLS C-SRC TYROSINE KINASE
SIGNALING AND REGULATES INNATE IMMUNITY
AND EPITHELIAL POLARITY IN CHOLANGIOCYTES
R. Fiorotto 1 , A. Villani 1 , R. Scirpo 2 , C. Spirli 1 ,
M. Strazzabosco 1,2
1 Department of Internal Medicine, Liver Center and
Digestive Diseases Section, Yale University, New
Haven, USA
2 Department of Surgery and Interdisciplinary
Medicine, University of Milano-Bicocca, Milan, Italy
CFTR is expressed at the apical membrane of cholangiocytes
where it regulates Cl− and HCO3 − secretion. CFTR also modulates
innate immune responses in the biliary epithelium. In fact, TLR4mediated responses to LPS are increased in cholangiocytes from
Cftrtm1Unc (Cftr-KO) mice along with the activity of c-Src, a nonreceptorial tyrosine kinase.
Aim: To understand how CFTR deficiency leads to up-regulation
of c-Src activity in cholangiocytes.
Methods and results: Primary cholangiocytes were isolated
from Cftr-KO mice and their WT littermates. In Cftr-KOcholangiocytes, c-Src inhibition by treatment with PP2 decreased
basal and LPS-stimulated NF-␬B activation and cytokines secretion.
CFTR co-immunoprecipitated with proteins involved in the negative regulation of c-Src (EBP-50, Csk and CBP); confocal imaging
confirmed their co-localization at the apical membrane in WT cells.
Since c-Src is important for maintaining the integrity of epithelial
cell–cell junctions, we investigated the distribution of ZO-1 (tight
junction), afadin (adherens junction) and subcortical F-actin fibers.
In Cftr-KO cells ZO-1 and afadin lost their junctional restriction and
also appeared diffusely distributed in the cytoplasm; also the cortical actin ring failed to form properly, suggesting a polarity defect.
Treatment with c-Src inhibitor PP2, rescued the polarity phenotype,
as shown by ZO-1 and F-actin re-distribution. Inhibition of c-Src
in vivo significantly attenuated biliary damage and inflammation
in a Cftr-KO mouse model.
Conclusions: Expression of CFTR is essential for the assembly of
an apical protein-complex located in lipid rafts that negatively regulates c-Src. Lack of CFTR perturbs this complex. Consequently c-Src
self-activates promoting an increase in TLR4 responses, the destabilization of cell–cell junctions and an impairment in cell polarity.
The protective effects of in vivo cSrc inhibition confirm the pathogenetic relevance of this mechanism and suggest that c-Src is a
potential therapeutic target in CFLD.
OC-05
OC-06
METFORMIN TARGETS A NEW
EZH2-PHOSPHOSTAT3-MIRNAS PATHWAY TO
INHIBIT LIPID DROPLETS ACCUMULATION AND
INTRACELLULAR INFLAMMATION
OMOMYC AGAINST HCC: A MYC DOMINANT
INTERFERING MINIPROTEIN TO COUNTERACT
TUMOR GROWTH
Pediconi 1,2 ,
Cocco 3 ,
Salerno 3 ,
N.
S. Di
D.
L. Belloni 2,3 , S. Piconese 3 , V. Barnaba 3 ,
M. Levrero 2,3
1
Department of Molecular Medicine, Sapienza
University, Rome, Italy
2 Sapienza Center for Life NanoScience, Rome, Italy
3 Department of Internal Medicine – DMISM,
Sapienza University, Rome, Italy
Background and aim: Accumulation of triglyceride-containing
lipid droplets (LDs) within hepatocytes in NAFLD patients is a
potentially reversible process, although sustained activation of
inflammatory signaling pathways leads to non-alcoholic steatohepatitis (NASH) that can evolve into cirrhosis and HCC. Here we
investigated the role of a new EZH2-phosphoSTAT3-miRNAs pathway in LD accumulation and intracellular inflammation in an in vitro
cellular model of vescicularsteatosis.
Methods: DMSO-differentiated human non-transformed HepaRG cells treated with oleic acid were used as a cellular model of
vescicolarsteatosis.
Results: dHepaRG cells treated with oleic acid show: (a) an
increased lipid accumulation and intracellular reactive oxygen
species (ROS) generation as compared to control cells; (b) deregulated lipid metabolism and liver-specific genes, including PDK4,
PLIN4, SLC2A1, ALB and ALDOB; (c) activation of an intracellular
inflammatory response, with the upregulation of IL6, IL8, OAS1,
NFKB and phosphoSTAT3. Oleate treatment also increased the
mRNA and protein levels of the EZH2 (Enhancer of Zeste Homolog
2) histone methyl-transferase that catalyzes the repressive histone
3 lysine 27 tri-methylation (H3K27me3). Treatment of oleateexposed HepaRG with S3I (STAT3 inhibitor) or MC1945 (new EZH2
inhibitor) reduces triglycerides accumulation, represses metabolic
genes over-expression and inhibits phosphoSTAT3 accumulation.
We also found that several STAT3/IL6 responsive miRNAs, including
miR21 and mir24, are upregulated after lipid overload, paralleling STAT3 activation. Chromatin immuno-precipitation (ChIP)
experiments showed that the oleate-dependent transcriptional
deregulation of these miRNAs correlates with H3K27me3 levels,
phosphoSTAT3 and EZH2 bound to their promoters. Moreover, metformin, an AMPK activator widely used as anti-diabetic drug and
known to improve lipid metabolism and to decrease steatosis in
animal models, reduces phosphoSTAT3, inhibits miRNAs upregulation and reduces triglyceride accumulation.
Conclusions: We show that a new EZH2-phosphoSTAT3miRNAs intracellular inflammation pathway amenable to therapeutic targeting is activated in well differentiated hepatocytes in
response to lipid overload and is involved in vescicularsteatosis.
e3
B. Barbaro 1,2 , C. Porcu 1 , G. Toietta 3 ,
R. Maggio 1,4 , M. Savino 4 , S. Nasi 4 , C. Balsano 4
1
Francesco Balsano Foundation, Rome, Italy
Department of Clinical and Applied Sciences and
Biotechnology, L’Aquila University, L’Aquila, Italy
3 Regina Elena Cancer Institute, Rome, Italy
4 IBPM – CNR, Rome, Italy
2
Hepatocellular carcinoma (HCC) is the fifth most common tumor
in the world for which the most promising strategy is targeted
therapy.
Abnormal Myc expression is linked to many solid tumors, but
this is usually not due to mutations in the Myc gene itself but results
from upstream mutations. Then our strategy is to counteract its
action avoiding its heterodimerization with Max, crucial for the
DNA binding and consequent gene expression activation. To this
aim we used a dominant negative molecule, termed OMOMYC.
To note, also Hedgehog pathway, mostly Gli1 target gene, is
reactivated during cancer, being normally switched-off in adults.
Deregulation of Myc and Gli1 pathways is frequently observed in
human hepatocarcinogenesis.
Aim: To determine whether HCC proliferation can be prevented
by Omomyc and to ascertain whether this strategy can affect the
Hedgehog pathway.
Methods: We engineered HepG2 and Huh7.5.1 human hepatoma cell lines to express Omomyc. We determined by microscope
examination, MTS assay and cytofluorimetric analysis, morphology, proliferation and viability of cells with or without Omomyc
induction. Real-time PCR and Western blot analysis were used for
determining the expression of Myc, PCNA (Proliferating Cellular
Nuclear Antigen), CCD1 (Cyclin D1), Hedgehog target gene Gli1.
Results: Omomyc is able to reduce HCC cell proliferation up to
70% in 8 days. Analysis of mRNA expression of genes involved in
cell proliferation and corresponding protein levels confirmed the
Omomyc effect in halting cell growth; moreover Omomyc was able
to decrease Gli1 expression.
Conclusion: Myc dominant negative miniprotein Omomyc is
able to prevent HCC proliferation in vitro by specifically counteracting the Hedgehog-mediated signaling.
We are now testing the outcome of this strategy in preventing
tumor progression in a xenograft mouse model of HCC, using cells
expressing Omomyc; we engineered the cells also with luciferase
to follow cancer growth in vivo by bioluminescence imaging.
e4
OC-07
OC-08
GUT–LIVER AXIS DERANGEMENT DUE TO LACK
OF INFLAMMASOME ACTIVITY LEADS TO
VISCERAL OBESITY AND NASH DEVELOPMENT
THE UK-PBC RISK SCORE: DERIVATION AND
VALIDATION OF A RISK SCORE TO PREDICT LIVER
EVENTS IN THE UK-PBC RESEARCH COHORT
L. Agostinelli 1 , C. Rychlicki 1 , E. Mingarelli 1 ,
C. Saponaro 2 , M. Gaggini 2 , E. Buzzigoli 2 ,
I. Pierantonelli 1 , S. Saccomanno 1 , C. Pinto 1 ,
L. Trozzi 1 , A. Benedetti 1 , S. De Minicis 1 ,
M. Marzioni 1 , A. Gastaldelli 2 , G. Svegliati-Baroni 1
M. Carbone 1,2,3 , S.J. Sharp 3 , M.A. Heneghan 4 ,
J.M. Neuberger 5 , G.M. Hirschfield 6 ,
A.K. Burroughs 7 , D. Thorburn 7 , A. Bathgate 8 ,
M. Aldersley 9 , C. Adgey 10 , P. Trembling 11 ,
K. Williamson 12 , L. Jopson 13 , R.T. Lim 5 ,
N.J. Wareham 3 , H.J. Cordell 14 , G.J. Alexander 1 ,
D.E. Jones 13 , R.N. Sandford 2 ,
G.F. Mells 1,2 , UK-PBC Consortium
1 Department of Gastroenterology, Polytechnic
University of Marche, Ancona, Italy
2 Institute of Clinical Physiology, National Research
Council, Pisa, Italy
Introduction: NAFLD, the most common form of chronic liver
disease, can lead to cirrhosis and hepatocellular carcinoma. Gut
dysbiosis and bacterial translocation induced by specific dietary
habits can elicit a proinflammatory and profibrogenic response.
NLRP3 inflammasome regulates intestinal homeostasis and mediates the release of proinflammatory cytokines in response to
cellular danger signals.
Aim: We want to investigate the role of NLRP3 inflammasome
in a Western-lifestyle diet model of NAFLD.
Methods: Wild-type (WT) C57BL/6 and Nlrp3A350VneoR
(Nlrp3−/− ) mice were fed either a chow diet or a high-fat diet
with fructose in drinking water (HFHC) for 12 weeks.
Results: Nlrp3−/− HFHC gained more weight (p < 0.01), showed
reduced energy expenditure and more fat mass (measured by doubly labeled water, all p < 0.05) with increased adipose tissue TNF␣
expression (p < 0.05), and developed more hepatic steatosis (higher
triglyceride content p < 0.01) compared to WT HFHC. Nlrp3−/−
HFHC showed increased intestinal permeability, (i.e., lower caecum zonulin-1 expression), that led to higher hepatic expression of
TLR4 (p < 0.01) and TLR9 (p < 0.05). No differences were observed
between Nlrp3−/− HFHC and WT HFHC in hepatic expression of
SREBP-1c effectors of de novo lipogenesis (ACC, FAS and SCD-1) that
were significantly increased in both groups. On the other hand,
Nlrp3−/− HFHC showed higher expression of PPAR2 (p < 0.01) and
of its downstream effectors FABP-4 (p < 0.05) and CD36 (p < 0.01)
that regulate lipid uptake and storage. In addition Nlrp3−/− HFHC
showed increased fatty acid mitochondrial-oxidation (i.e., higher
CPT1A expression) that, associated to the reduced expression
of NRF2 transcription-factor, led to increased superoxide production (measured by dihydroethidium-staining) (all p < 0.01).
These events were associated to increased macrophage infiltration, collagen␣1(I) and MCP1 gene expression (p < 0.05) in Nlrp3−/−
HFHC mice only.
Conclusions: In the Western-lifestyle diet, lack of NLRP3 inflammasome is associated with bacterial products translocation that
leads to metabolic alterations in both adipose tissue and liver, and
to NASH development.
1 Division of Gastroenterology and Hepatology,
Addenbrooke’s Hospital, Cambridge, United Kingdom
2 Academic Department of Medical Genetics,
University of Cambridge, Cambridge, United
Kingdom
3 MRC Epidemiology Unit, University of Cambridge,
Cambridge, United Kingdom
4 Institute of Liver Studies, King’s College Hospital,
London, United Kingdom
5 Liver Unit, Queen Elizabeth Hospital, United
Kingdom
6 NIHR Biomedical Research Unit and Centre for Liver
Research, University of Birmingham, Birmingham,
United Kingdom
7 The Sheila Sherlock Liver Centre, The Royal Free
Hospital, London, United Kingdom
8 Liver Unit, Royal Infirmary Hospital, Edinburgh,
United Kingdom
9 Liver Unit, St James’s Hospital, Leeds, United
Kingdom
10 Liver Unit, Royal Victoria Hospital, Belfast, United
Kingdom
11 Liver Unit, Barts and The London NHS Trust,
London, United Kingdom
12 Translational Gastroenterology Unit, John
Radcliffe Hospital, Oxford, United Kingdom
13 Institute of Cellular Medicine, Newcastle
University, Newcastle, United Kingdom
14 Institute of Genetic Medicine, Newcastle
University, Newcastle, United Kingdom
Introduction: Outcomes in primary biliary cirrhosis (PBC) can
be predicted by biochemical response to ursodeoxycholic acid
(UDCA). However, existing definitions do not take the stage of the
liver disease into account and dichotomize UDCA response and
long-term risk whereas both are a continuum. We analyzed the
UK-PBC Research Cohort to develop a risk score that includes markers of disease stage, as well as post-treatment liver biochemistries
modeled as continuous variables.
Methods: We constructed a PBC risk score for LT and liverrelated death at 15 years in a derivation cohort (N = 2422) and
evaluated it in a validation cohort (N = 1600). We used multivariable
fractional polynomials (MFP) to model non-linear risk relations
with continuous variables, and 20 multiple imputations to replace
missing values. We fit a Cox proportional hazards model in each
imputed dataset, and used Rubin’s rules to combine the results.
The resulting coefficients were used together with the baseline survivor function to derive an equation for absolute risk at 15 years.
Net reclassification improvement (NRI) was calculated to compare
the predictive performance of this risk score compared with other
prognostic scores.
Results: Albumin and platelet at baseline, ALP, bilirubin and
transaminases after 12 months of UDCA were independently associated with a liver event. The score incorporated these variables,
appropriately MFP-transformed. When applied to the validation
cohort, the score was highly predictive (c-statistic = 0.90). The NRI
showed that the risk score had greater ability to identify individuals with and without events compared to other risk scores (NRI
of PBC risk score vs. Paris1 = 90%; Paris2 = 64%; Barcelona = 38%;
Toronto = 71%).
Conclusions: Prognosis of UK patients with PBC can be accurately assessed with the PBC risk score by using readily available
objective clinical measures. This may be used to identify high-risk
patients for closer monitoring and second-line therapies, as well
as low-risk patients who require infrequent monitoring and might
even be followed-up in primary care.
OC-09
LIVER DAMAGE CAN BE ASSOCIATED WITH
DEREGULATION OF THE DE NOVO LIPOGENESIS
PATHWAY IN SUBJECTS WITH NON ALCOHOLIC
FATTY LIVER DISEASE
C. Rosso 1 , C. Saponaro 2,3 , L. Mezzabotta 1 ,
E. Vanni 1 , R. Gambino 1 , F. Saba 1 ,
R. Ibrahim Kamal Jouness 1 , M. Gaggini 2,4 ,
E. Buzzigoli 2 , G.P. Caviglia 1 , M.L. Abate 1 ,
F. Salomone 5 , A. Smedile 1 , M. Rizzetto 1 ,
M. Cassader 1 , A. Gastaldelli 2 , E. Bugianesi 1
1
Department of Medical Sciences, University of
2 Cardiometabolic Risk Unit, CNR-Institute of Clinical
Physiology, Pisa, Italy
3 Dottorato Pegaso Regione Toscana in Biochimica e
Biologia Molecolare, University of Siena, Siena, Italy
4 Dipartimento di Patologia Chirurgica, Molecolare e
di Area Critica, University of Pisa, Pisa, Italy
5 U.O.C. of Gastroenterology, Azienda Sanitaria
Provinciale di Catania, Catania, Italy
Introduction: Adipose tissue insulin resistance (IR) and elevated adipose free fatty acid (FFA) flux are prominent features in
NAFLD. De novo lipogenesis (DNL) is up to 3-fold higher in subjects with fatty liver and is not suppressed on fasting. When DNL
is stimulated, the production of saturated FAs is increased and the
oxidation of FAs of any source is reduced. Both mechanisms can
favor inflammation and IR.
Aim: We measured FFA flux/composition and used a surrogate
index of DNL (DNLi) to evaluate their relationship with histological
features in a group of non-diabetic subjects with biopsy-proven
NAFLD.
Materials and methods: Hepatic and adipose tissue-IR indices
were derived from [2H5]glycerol and [2H2] glucose kinetics in a
group of non-diabetic NAFLD patients in the basal state (n = 40)
and after a 4 h oral glucose load test (n = 20). Gas chromatography
mass spectrometry was used to assess FFAs composition. DNLi was
derived as the ratio palmitic/linoleic acid.
Results: Fasting plasma glucose/insulin, lipid profile, hepatic/adipose tissue IR indices and DNLi were similar in the two
groups. Fasting DNLi was associated with triglycerides (TG) and
FFAs levels and with adipose tissue IR (r = 0.597, r = 0.330 and
r = 0.394, respectively). Among histological features, fasting DNLi
significantly correlated with steatosis (r = 0.364, P = 0.02) and NAS
score (r = 0.306, P = 0.05). After the glucose load, TG levels initially
e5
increased despite elevated insulin levels, suggesting a significant
contribution of DNL. Accordingly, DNLi consensually increased with
TG levels (r = 0.749, P < 0.005) and was significantly related to the
degree of fibrosis (rs = 0.514, P = 0.02).
Conclusions: Oral glucose load is associated with changes in
DNL and hepatic triglyceride synthesis that can favor liver fibrosis
in patients with NAFLD.
Funding: Funded by FP7/2007-2013 under grant agreement
n◦ HEALTH-F2-2009-241762 for the project FLIP and by PRIN
2009ARYX4T.
OC-10
THE PRESENCE OF WHITE MATTER LESIONS IS
NOT ASSOCIATED WITH NON-ALCOHOLIC FATTY
LIVER DISEASE BUT WITH ITS HISTOLOGICAL
SEVERITY
S. Petta 1 , A. Tutolomondo 2 , C. Gagliardo 4 ,
R. Zafonte 5 , G. Brancatelli 4 , D. Cabibi 3 ,
C. Cammà 1 , V. Di Marco 1 , L. Galvano 5 , A. Licata 2 ,
F. Magliozzo 5 , G. Merlino 5 , M. Midiri 4 , A. Pinto 2 ,
A. Craxì 1
1 Sezione di Gastroenterologia, DiBiMIS, University of
Palermo, Italy
2 Dipartimento Biomedico di Medicina Interna e
Specialistica (Di.Bi.M.I.S), Università di Palermo,
Palermo, Italy
3 Cattedra di Anatomia Patologica, University of
Palermo, Palermo, Italy
4 Department of Radiology, University of Palermo,
Palermo, Italy
5 Medicina Generale Palermo, Palermo, Italy
Background: Nonalcoholic fatty liver disease (NAFLD) has been
associated with increased cardiovascular risk, including coronary
artery disease and cerebrovascular events. No studies however
assessed the potential relationship between NAFLD and subclinical cerebrovascular alterations. We tested the correlation between
NAFLD and its histological severity with vascular white matter
lesions (WML) in patients with biopsy-proven NAFLD and in non
steatosic controls.
Methods: The anthropometric, biochemical and metabolic features were recorded in 77 consecutive biopsy-proven NAFLD
(Kleiner score), and in 35 controls with normal ALT, without chronic
liver diseases, and without ultrasonographic evidence of steatosis.
All patients underwent magnetic resonance assessment of WML.
WML were classified according to the Fazekas score in absent (0/III),
or present (mild I/III; moderate II/III, and severe I/III). For purpose of
analyses all controls, as plausible, were considered without NASH
and without F2-F4 liver fibrosis.
Results: WML were found in 26% of the entire cohort (29/112),
even if of a moderate–severe grade in 5 patients only. The prevalence of WML was similar in NAFLD compared to no NAFLD (27% vs
23%; p = 0.62). Age ≥50 yrs, female gender, type 2 diabetes, arterial
hypertension, presence of NASH (35% vs 18%, p = 0.05) and presence
of F2-F4 fibrosis (43% vs 17%, p = 0.003) were associated with WML
presence (p = <0.01). At multivariate analysis age >50 yrs (OR 3.44
95% CI 1.01–11.6. p 0.04), female gender (OR 3.71. 95% CI 1.28–10.7.
p 0.01), and F2-F4 fibrosis (OR 3.39. 95% CI 1.17–9.84. p 0.02) were
maintained as factors independently associated with WML. When
considering NAFLD patients only, we confirmed F2-F4 fibrosis as
the only independent predictor of WML (OR 4.24. 95% CI 1.14–15.7.
p 0.03).
e6
Conclusion: The presence of WML is not associated with NAFLD
but with its histological severity. Clinical implications of this issue
need to be assessed by longitudinal studies.
OC-11
LONG-TERM USE OF HUMAN ALBUMIN FOR THE
TREATMENT OF ASCITES IN PATIENTS WITH
HEPATIC CIRRHOSIS: THE INTERIM ANALYSIS OF
THE ANSWER STUDY
M. Bernardi 1 , O. Riggio 2 , P. Angeli 3 ,
C. Alessandria 4 , S. Neri 5 , F.G. Foschi 6 ,
F. Levantesi 7 , S. Boccia 8 , A. Airoldi 9 , S. Fagiuoli 10 ,
G. Svegliati-Baroni 11 , G. Laffi 12 , R. Cozzolongo 13 ,
G. Butera 14 , V. Sangiovanni 15 , P. Toniutto 16 ,
M.A. Zocco 17 , R. De Marco 18 , F. Morisco 19 ,
F. De Leonardis 20 , I. Cacciola 21 , G. Elia 22 ,
A. Federico 23 , S. Massironi 24 , R. Guarisco 25 ,
A. Marin 26 , S. Piano 3 , C. Elia 4 , S. Nardelli 2 ,
D. Maiorca 5 , E. Neri 6 , A. Mastroianni 7 ,
M. Tufoni 1 , L. Simone 8 , L. Cesarini 9 , G. Magini 10 ,
M. Marzioni 11 , R.G. Romanelli 12 ,
M. Zappimbulso 13 , F. Macaluso 14 , G. Parrella 15 ,
F. Pugliese 16 , A. Tortora 17 , M. Cavallin 3 ,
A. Andrealli 4 , C. Pasquale 2 , F. Fidone 5 , A. Lanzi 6 ,
A. Alberti 9 , F. Salerno 27 , A. Roncadori 28 ,
G. Zaccherini 1 ,
P. Caraceni 1 , the ANSWER Study Group
1
Università di Bologna, Italy
Università di Roma “La Sapienza”, Italy
3 Università di Padova, Italy
4 A.O. Città della Salute e della Scienza di Torino, Italy
5 Università di Catania, Italy
6 A.U.S.L. Ravenna – Osp. di Faenza, Italy
7 A.U.S.L. Bologna – Osp. di Bentivoglio, Italy
8 A.O.U. di Ferrara, Italy
9 A.O. Niguarda Ca’ Granda di Milano, Italy
10 A.O. Papa Giovanni XXIII di Bergamo, Italy
11 Università di Ancona, Italy
12 Università di Firenze, Italy
13 IRCSS “De Bellis” di Castellana Grotte, Italy
14 Università di Palermo, Italy
15 A.O.R.N. dei Colli – Osp. Cotugno di Napoli, Italy
16 Università di Udine, Italy
17 Università Cattolica di Roma, Italy
18 A.O. di Cosenza, Italy
19 Università di Napoli “Federico II”, Italy
20 Università di Roma “Tor Vergata”, Italy
21 Università di Messina, Italy
22 A.O.U. di Parma, Italy
23 Seconda Università di Napoli, Italy
24 Università di Milano – Osp. Maggiore Policlinico,
Italy
25 A.S.L. Roma H – Osp. “San Giuseppe” di Marino,
Italy
26 A.U.L.S.S. Mirano – Osp. di Dolo, Italy
27 Università di Milano – Policlinico San Donato, Italy
28 C.I.N.E.C.A. – Bologna, Italy
Methods: In this multicentre, prospective, randomized clinical
trial, 420 patients with cirrhosis and uncomplicated ascites treated
with anti-mineralocorticoids (≥200 mg/day) and furosemide
(≥25 mg/day) are planned to be randomized 1:1 to either standard
medical treatment (SMT) or SMT + HA (40 g twice weekly for 2
weeks, and then 40 g weekly). Death, liver transplantation, TIPS,
refractory ascites requiring ≥3 paracentesis/month or 18 months
follow-up terminate the study. The primary end-point is mortality.
Among the secondary end-points, those related to the management of ascites and the incidence of complications of cirrhosis were
assessed in this analysis.
Results: 386 (SMT: 188; SMT + HA: 198) patients were included.
Their respective median follow-up duration was 183 (129–233)
and 301 (238–355) days (p = 0.021). Baseline demographics and
clinical and laboratory parameters were well balanced between
the two arms. Kaplan–Meier intention-to-treat analysis showed
that mortality was significantly reduced in patients receiving
HA (at 18 months: SMT + HA: 22%, SMT: 34%; p = 0.045). Statistically significant benefits were found in the SMT + HA arm in the
incidence rate of paracentesis (−55%, p < 0.001), incidence of refractory ascites (−42%, p < 0.001), and need of ≥3 paracentesis/month
(−62%; p < 0.001). HA arm also presented an advantage in the
incidence rates of SBP (−57%; p = 0.004), hepatic encephalopathy
grade III–IV (−37%; p = 0.005) and renal impairment (serum creatinine > 1.5 mg/dl) (−28%; p = 0.011). HA administration did not
increase the risk of variceal bleeding, and the incidence of severe
adverse effects was similar in the two arms.
Conclusions: Long-term HA administration prolongs survival in
patients with cirrhosis and ascites, improves greatly the management of ascites and reduces the incidence of severe complications
of the disease.
2
Introduction: Despite long-term human albumin (HA) administration for treating cirrhosis with ascites is commonly used in Italy,
the scientific evidence of its efficacy is still lacking.
Aim: To assess the efficacy of long-term HA administration in
patients with cirrhosis and ascites.
OC-12
VALIDATION OF A NEW CHILD-TURCOTTE-PUGH
CLASS 0 FOR PATIENTS WITH HEPATOCELLULAR
CARCINOMA IN A EUROPEAN COHORT
A. Gianstefani, A. Pecorelli, I. Pettinari,
L. Venerandi, F. Piscaglia,
L. Bolondi, the ITA.LI.CA. Study Group
Department of Medical and Surgical Sciences,
S.Orsola-Malpighi Hospital, Alma Mater Studiorum –
University of Bologna, Bologna, Italy
Background: Hepatocellular carcinoma (HCC) often arises in
the context of very well-preserved liver function, expressed by
a Child-Turcotte-Pugh (CTP) class A, which comprises only two
scores, 5 and 6. Recent published data identified a new CTP
score, named 0, defined by fulfilling all the subsequent criteria:
albumin ≥ 4 g/dL, bilirubin ≤ 0.8 mg/dL, prothrombin time prolongation < 0 s, no ascites, no encephalopathy. In an Asiatic population,
this subgroup of HCC patients with very well-preserved liver function had a better outcome compared to the CTP class A patients.
Aim: To validate the prognostic role of the new CTP 0 in a European cohort of HCC patients.
Methods: A total of 4248 patients with a first diagnosis of
HCC enrolled in the ITA.LI.CA. database between 1986 and 2012
were retrospectively analyzed. Patients were divided in four groups
according to the CTP class: 0 (232 pts), A (2514 pts), B (1221 pts)
and C (281 pts). Patient survival probability was estimated using
Kaplan–Meier method.
Results: The 8.4% of CTP A patients were reclassified as CTP
0. Median overall survivals statistically differed among groups
(months; 95% CI): CPT 0 62 (52.9–71.1), A 44 (41.6–46.4), B 22
(19.7–24.3), C 9 (6.6–11.3), p < 0.0001. Comparisons between survivals of CTP 0 vs A, B and C were also statistically different
(p < 0.0001 in all associations). The prognosis of patients in the
intermediate BCLC stage also differed according to the liver function
(0 vs A vs B, p < 0.0001).
Conclusions: The newly proposed CTP class 0 identifies a different subgroup of patients with a better prognosis, also when applied
in a European cohort, where HCV aetiology is predominant.
This new approach impacts not only on outcome prediction but
also, potentially, on treatment allocation, better stratifying, in particular, the heterogeneous intermediate BCLC-B stage.
OC-13
PATHOGENESIS AND CLINICAL IMPACT OF
RELATIVE ADRENAL INSUFFICIENCY IN
HOSPITALIZED PATIENTS WITH ACUTE
DECOMPENSATION OF CIRRHOSIS
S. Piano 1,2 , E. Favaretto 2 , S. Fasolato 1,2 ,
C. Scaroni 2 , E. Gola 2 , A. Brocca 2 , A. Sticca 2 ,
F. Morando 2 , M. Cavallin 2 , A. Romano 2 ,
E. Gringeri 3 , A. Gatta 2 , U. Cillo 3 , M. Plebani 2 ,
P. Angeli 1,2
1 Unit of Hepatic Emergencies and Liver
Transplantation, University of Padua, Padua, Italy
2 Department of Medicine, University of Padua,
Padua, Italy
3 Unit of Hepatobiliary Surgery and Liver
Background and aims: Relative adrenal insufficiency (RAI) has
been described in patients with cirrhosis and has been recently
associated with the development of hepatorenal syndrome, sepsis,
and poor survival. Its pathogenesis has not yet been clearly defined.
The aim of our study was to explore factors associated with RAI and
to investigate clinical impact of RAI in these patients.
Methods: 94 patients admitted to the hospital for an acute
decompensation of cirrhosis were consecutively enrolled in the
study and followed up for 90 days. Adrenal function was assessed
with short synacthen test (SST). RAI was diagnosed when the
increase in serum total cortisol after SST was <9 ␮g/dL in patients
with basal serum total cortisol < 35 ␮g/dL. Bacterial infections,
markers of inflammations, ACTH, substrates for steroidogenesis
and apolipoprotein A1 (ApoA1), which is required for cholesterylester accumulation in steroidogenic cells, were explored as possible
pathogenetic factors involved in RAI.
Results: RAI was diagnosed in 42.6% of patients. Those with
RAI were younger (57.0 vs 61.5 years; p = 0.047), had higher
MELD-Na score (22 vs 18.4; p = 0.01), higher C-reactive protein
(15.0 vs 11.5 mg/L; p = 0.047), and lower total cholesterol (1.5
vs 2.1 mmol/L; p = 0.028), HDL (0.4 vs 0.6 mmol/L; p = 0.034) and
apolipoprotein A1 (0.6 vs 0.8; p = 0.006) than those without RAI.
TNF-alpha, IL6, IL-10, IL1 beta and ACTH were not significantly
different between the two groups. ApoA1 was the only independent predictor of RAI (OR = 0.12; p = 0.011). In our series RAI
was associated with a higher risk to develop bacterial infections
(65.1 vs 42.4%; p = 0.039), hyponatremia (46.0 vs 14.3%; p = 0.002),
and with poorer 90-day transplant free survival (70.7 vs 90.3%;
p = 0.013). In multivariate analysis MELD-Na (HR = 1.096; p = 0.030),
age (HR = 1.059; p = 0.043) and RAI (HR = 3.277; p = 0.041) were
found to be independent predictors of mortality.
e7
Conclusions: RAI is frequent in patients hospitalized for an
acute decompensation of cirrhosis. Low level of ApoA1 seems to
have a pivotal role in its pathogenesis. RAI is associated with a high
risk to develop bacterial infections, hyponatremia and with a low
probability of survival in these patients.
OC-14
COST-EFFECTIVENESS OF PRE-TRANSPLANT
SOFOSBUVIR TO PREVENT RECURRENCE OF HCV
INFECTION AFTER LIVER TRANSPLANTATION
A. Vitale 1 , G. Spolverato 1 , P. Burra 1 ,
T.M. De Feo 2 , U. Cillo 1 , S. Fagiuoli 3 , On behalf of
the Liver Transplantation NITp Working Group
1
University Hospital of Padua, Padua, Italy
North Italy Transplant Program, Fond. IRCCS Ca’
Granda OMP, Milan, Italy
3 Gastroenterology and Transplant Hepatology,
Ospedale Papa Giovanni XXIII, Bergamo, Italy
2
Background and aims: The strong efficacy of pre-transplant
Sofosbuvir (SOF) plus Ribavirin in preventing recurrence of hepatitis C virus (HCV) infection after liver transplantation (LT) has
been reported. The aim of this study was to evaluate the costeffectiveness of this strategy in the North Italy Transplant program
(NITp) area.
Methods: We first evaluated the impact of HCV infection on
post-LT survival in 2376 consecutive adult patients (MELD ≤ 25,
period 2004–2009, NITp area) and prevalence-costs of conventional standard of care (SOC) antiviral therapy (Peginterferon plus
ribavirin) after LT. A Markov model was developed to compared
two strategies: SOF + Ribavirin (RBV) as pre-LT anti-HCV treatment (strategy A), versus on-demand post-LT SOC antiviral therapy
(strategy B). The endpoint was incremental cost-effectiveness ratio
(ICER) as costs ($)/quality-adjusted life year (QALY).
Results: Among the 1794 patients undergoing LT and meeting the inclusion criteria, 860 (48%) were HCV+ and 50% of them
received SOC therapy post-LT (drugs and adverse effects management mean costs = 16,440$ for patient). HCV etiology had a strong
impact on post-LT survival (Hazard Ratio = 1.59, 95% CI = 1.22–2.09,
p = 0.0007). After Monte Carlo simulation, pre-LT SOF therapy
showed a median survival benefit of 2.0 QALYs, and an ICER of
33,600$/QALY. Costs of SOF therapy, sustained viral response rate
12 weeks after LT, and recipient age were the main ICER predictors
at multivariate analysis.
Conclusions: Pre-transplant SOF in HCV patients proved to be
a cost-effective treatment compared to post-LT SOC therapy.
e8
OC-15
OC-16
ENOS POLYMORPHISMS IN RELATION TO
OUTCOME IN ADVANCED HCC PATIENTS
RECEIVING SORAFENIB
THE AAA+ ATPASE RUVBL1 COORDINATES LIVER
METABOLISM AND HEPATOCELLULAR
CARCINOMA PROGRESSION
A. Casadei Gardini 1,∗ , G. Marisi 1 , E. Scarpi 1 ,
M. Scartozzi 2 , F.G. Foschi 3 , L. Faloppi 3 , E. Tenti 1 ,
S. Tamberi 3 , E. Tamburini 3 , G.L. Frassineti 1 ,
S. Cascinu 2 , P. Ulivi 1
T. Mello 1,2 , M. Materozzi 1 , F. Zanieri 1 ,
O. Bereshchenko 3 , E. Ceni 1 , M. Tarocchi 1,2 ,
G. Marroncini 1 , I. Simeone 1 , S. Polvani 1 ,
S. Tempesti 1 , C. Nerlov 4 , S. Milani 1,2 , A. Galli 1,2
1
1
IRST-IRCCS, Meldola, Italy
Department of Medical Oncology, A.O. Ospedali
Riuniti Università Politecnica delle Marche, Ancona,
Italy
3 Ospedale per gli Infermi, Faenza, Italy; AUSL
Ravenna, Faenza, Italy
2
Background and aims: Cancer cells adapt to hypoxic microenvironment through the activation of many molecules, including
endothelial nitric oxide synthase (eNOS). Sorafenib, by blocking the
vascular endothelial growth factor receptors (VEGFRs), induces an
inhibition of eNOS activity with a consequent decrease of the production of nitric oxide (NO). NO is associated with an increase of
tumor angiogenesis, tumor invasion and metastasis formation. In
our study we analysed the role of eNOS polymorphisms in relation to clinical outcome in patients with hepatocellular carcinoma
treated with sorafenib.
Methods: From a database of 257 patients diagnosed with hepatocellular carcinoma from 2004 to 2014, we selected 54 patients
who received sorafenib. Peripheral blood samples or FFPE tumor
tissues were available for DNA extraction and genotyping analysis. Three eNOS polymorphisms (eNOS +894 G/T, eNOS VNTR 27 bp
4a/b, eNOS −786 C/T) were analyzed by direct sequencing or Real
Time PCR method. We analyzed 21 patients for the VNTR 4a/b polymorphism and 32 patients for −786 C/T polymorphism. All the
candidate genotypes were evaluated to identify a potential correlation with overall survival (OS) (log-rank test).
Results: With regard to eNOS VNTR it was observed that patients
carrying the b allele (5 repetitions of 27 bp) both in homozygosity
(4bb) and in heterozygosity (4ab) were associated with a better
OS. The variants 4aa (4 repeats of 27 bp in homozygosity), 4ab
and 4bb, were associated with a median OS of 5.7, 13.9 and 23.6
months, respectively (p = 0.016). For eNOS-786 the presence of the
T allele both in homozygosity (TT) and in heterozygosity (TC) was
associated with a statistically significant longer OS with respect
to patients with CC genotype (15.6 versus 13.9 months, respectively, p = 0.031). No correlations were observed in relation to PFS
(p = 0.494).
Conclusions: eNOS VNTR and eNOS −786 could represent
prognostic markers in patients with advanced hepatocellular carcinoma treated with sorafenib.
Department of Biomedical, Experimental and
Clinical Sciences “Mario Serio”, University of
Florence, Florence, Italy
2 Careggi University Hospital, Florence, Italy
3 Department of Medicine, Section of Pharmacology,
University of Perugia, Perugia, Italy
4 Institute for Stem Cell Research and MRC Centre for
Regenerative Medicine, University of Edinburgh,
Edinburgh, UK
The AAA+ ATPase Ruvbl1 is overexpressed in several human
cancers, including hepatocellular carcinoma (HCC), in which high
Ruvbl1 expression correlates with a poor prognosis. A growing body
of data from in vitro models supports the concept that dysregulation
of Ruvbl1 expression occurs in cancer to promote its growth and
progression, making this protein an attractive target for anti-cancer
therapies. However, whether Ruvbl1 participate in the oncogenic
transformation and cancer progression in vivo remains speculative.
To challenge this question we realized a hepatocyte-conditional
Ruvbl1 hemizygous mouse and evaluated the tumor onset and progression after DEN injection.
Ruvbl1 hemizygous mice were obtained by crossing Ruvbl1floxed with Albumin-Credeleter mice. The male offspring were
subjected to a single i.p. injection of DEN (5 mg/kg) to induce liver
cancer, and were monitored up to one year after cancer induction.
Conditional liver-hemizygous mice showed no obvious phenotype with respect to liver size, viability and liver function (AST-ALT),
however they had significant alterations of glucose, triglycerides
and cholesterol serum levels, and an increased body weight.
Despite a significant delay in the onset of liver cancer, Ruvbl1+/−
mice eventually developed significantly larger tumors than control mice. We found that Ruvbl1+/− mice had reduced hepatocyte
turnover and impaired mTOR signaling than wild-type mice, which
is likely causing both the metabolic alterations and the delayed
onset of HCC.
In conclusion, this is the first report highlighting a role of
Ruvbl1 as a major regulator of hepatic metabolism. Contrary to our
expectations, we found that although in the hepatocyte-conditional
Ruvbl1+/− mice the onset of HCC is delayed, its progression
is accelerated. The underlying molecular mechanisms are under
investigation, nevertheless this report highlights the potential risks
of prolonged Ruvbl1 inhibition in a intact mammalian organism.
Acknowledgement: This research is funded by the Italian Ministry of Health through grant GR-2009-1600315.
e9
OC-17
OC-18
THE EMPIRICAL ANTIBIOTIC TREATMENT OF
NOSOCOMIAL SPONTANEOUS BACTERIAL
PERITONITIS IN PATIENTS WITH
DECOMPENSATED CIRRHOSIS: RESULTS OF A
RANDOMIZED CONTROLLED CLINICAL TRIAL
RISK FACTOR FOR EARLY KIDNEY DISEASE
AFTER LIVER TRANSPLANTATION AND ITS
IMPACT ON GRAFT LOSS: INSIGHTS FROM THE
LIVER MATCH COHORT STUDY
S. Piano 1,2 , F. Salinas 3 , S. Fasolato 1,2 ,
F. Morando 2 , M. Cavallin 2 , A. Romano 2 , S. Rosi 2 ,
M. Stanco 2 , A. Sticca 2 , M. Senzolo 4 , P. Burra 4 ,
G. Zanus 5 , U. Cillo 5 , A. Gatta 2 , P. Angeli 1,2
Padua, Italy
3 Division of Medicine, Private Hospital “Giovanni
XXIII” of Monastier, Treviso, Italy
4 Multivisceral Transplant Unit, University of Padua,
Padua, Italy
5 Unit of Hepatobiliary Surgery and Liver
Background and aims: Spontaneous bacterial peritonitis (SBP)
is a common and life-threatening complication of cirrhosis. Third
generation cephalosporins (TGCs) are the first line empirical treatment of SBP. In recent years, an increasing rate of SBP due to TGCs
resistant bacteria has been observed, in particular in nosocomial
episodes. Currently a broader spectrum antibiotic regimen such as
carbapenems and glycopeptides/lipopeptides has never been compared to TGCs in the treatment of nosocomial SBP. The aim of our
study was to compare the efficacy of meropenem plus daptomycin
versus ceftazidime in the treatment of nosocomial SBP.
Methods: Patients with cirrhosis, ascites and nosocomial SBP
were randomized to receive meropenem (1 g/8 h) plus daptomycin
(6 mg/kg/day) or ceftazidime (2 g/8 h) plus albumin in both groups.
A diagnostic paracentesis was performed after 48 h of antibiotic
treatment. A reduction in ascitic fluid neutrophil count <25% of the
pre-treatment value was considered a treatment failure and antibiotic therapy was changed accordingly. The primary outcome was
the efficacy of the treatment defined by the resolution of SBP after
7 days of treatment (NCT01455246).
Results: 32 patients were randomized and 31 were analyzed.
The combination of meropenem plus daptomycin was significantly
more effective than ceftazidime in the treatment of nosocomial
SBP (86.7 vs 25%; p < 0.001). TGCs resistant bacteria were isolated
in 81.3% of positive cultures. In multivariate analysis, ineffective response to first line treatment (hazard ratio [HR] = 20.6;
p = 0.01), development of acute kidney injury during hospitalization
(HR = 23.2; p = 0.01) and baseline mean arterial pressure (HR = 0.92;
p = 0.01) were found to be independent predictors of 90-day transplant free survival. The incidence of adverse events was similar
between the two groups.
Conclusions: The combination of meropenem plus daptomycin
is more effective than ceftazidime in the treatment of hospital
acquired SBP. The efficacy of the first line treatment is associated
with improved 90-day survival in these patients.
S. Ginanni Corradini 1 , A.P. Mitterhofer 2 ,
A. Nardi 3 , T. Marianelli 4 , L. Parlati 1 , F. Tinti 2 ,
M. Angelico 4 , the Liver Match Study Group
1
Gastroenterology, Sapienza Università di Roma,
Italy
2 Nephrology, Sapienza Università di Roma, Italy
3 Biostatistics, Università di Tor Vergata, Roma, Italy
4 Hepatology and Transplant, Università di Tor
Vergata, Roma, Italy
Introduction and aim: Early Kidney Dysfunction (EKD) is frequent after liver transplantation (LT). We investigated the risk
factors for EKD and its effect on graft loss.
Methods: We evaluated 1302 patients undergoing LT in Italy
from June 1, 2007 to May 31, 2009 belonging to the Liver
Match cohort. EKD was staged based on the lowest estimated
glomerular filtration rate (eGFR, MDRD-4 formula) at 3 and 6
months after LT, as follows: eGFR > 60 ml/min (EKD stage 1-2);
45 ≤ eGFR < 60 ml/min (EKD stage 3a); 30 ≤ eGFR < 45 ml/min (EKD
stage 3b); and eGFR < 30 ml/min (EKD stage 4–5).
Results: 812 patients had EKD stage 1–2 (62.4%), 404 EKD stage
3a/b (31.0%) and 86 EKD stage 4–5 (6.6%). Multinomial logistic analysis identified four recipient risk factors for EKD: old age, female
gender, and low eGFR and sodium levels at LT (table).
EKD stage 3a/b vs stage 1–2
Odds ratio
Recipients age
(×10 years)
Recipient gender
(F vs M)
eGFR at LT 30–60
vs ≥60 ml/min
eGFR at LT < 30 vs
≥60 ml/min
Recipient sodium
at LT (per
5 meq/L)
95% C.I.
EKD stage 4–5 vs stage 1–2
p-Value
Odds ratio
95% C.I.
p-Value
1.63
1.40–1.89
<0.0001
1.70
1.28–2.27
0.0003
2.29
1.68–3.12
<0.0001
2.00
1.58–3.46
0.0129
2.55
1.79–3.64
<0.0001
2.72
1.50–4.94
0.0010
2.82
1.35–5.89
0.0059
9.74
4.08–23.28
0.86
0.75–0.98
0.0252
0.73
0.58–0.92
<0.0001
0.0082
Multivariable Cox regression identified EKD stage as a significant risk factor for 5-year graft loss. Assuming EKD stage 1–2
as reference level, the hazard ratio of graft loss for EKD stage
3a, 3b, and 4–5 were 0.89 (p = 0.4729), 1.68 (p = 0.0059) and 3.38
(p < 0.0001), respectively.
Conclusions: The probability of EKD can be estimated before
LT and EKD stage > 2b entails a high risk of graft loss. Patients at
high risk for EKD should be considered for alternative treatment
strategies, including combined liver-kidney transplantation.
e10
OC-19
OC-20
PREVENTION OF BLEEDING FOLLOWING
INVASIVE PROCEDURES IN CIRRHOTIC
PATIENTS: A SINGLE CENTER EXPERIENCE
ANTI CAPSID DRUGS HAP12 AND AT130 TARGET
HBV CORE PROTEIN NUCLEAR FUNCTIONS
G. Tosetti 1 , F. Invernizzi 1 , V. La Mura 1 ,
A. Aghemo 1 , M. Primignani 1 , A. Sangiovanni 1 ,
M.F. Donato 1 , A. Nicolini 2 , M. Iavarone 1 ,
M. Colombo 1
1
Division of Gastroenterology and Hepatology,
Fondazione IRCCS Cà Granda Ospedale Maggiore
Policlinico, University of Milan, Milan, Italy
2 Division of Radiology, Fondazione IRCCS Cà Granda
Ospedale Maggiore Policlinico, University of Milan,
Milan, Italy
Background: Risk of bleeding in cirrhosis has predominantly
been associated with coagulopathy and thrombocytopenia due to
impaired liver function and splenomegaly. The evaluation of the
actual bleeding risk in cirrhotic patients undergoing invasive procedures is a critical point to optimize management in terms of platelet
(Plt) or plasma prophylactic transfusions.
Methods: During 2013, 480 cirrhotic patients underwent
diagnostic or therapeutic invasive procedures at our Liver Unit
(Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico,
Milan, Italy). Plt transfusion was performed when Plt count less
than 50,000/mmc while fresh frozen plasma was infused when
international normalized ratio (INR) > 1.5, except those undergoing
paracentesis. Low-molecular-weight-heparin was discontinued
24 h before any procedure while antiaggregant therapy was interrupted five days before any procedure except oesophageal varices
band ligation and paracentesis. Major hemorrhagic events were
those requiring hospitalisation or blood transfusion; minor events
was a haemoglobin decline > 1.5 g/dl post procedure without clinical relevance.
Results: 174 Transarterial Chemo-Embolization (TACE), 16
Radio-Frequency Termal Alblation (RFTA), 214 paracentesis, 59
oesophageal varices banding, 6 trans-jugular liver biopsy (LB) and
11 Percutaneous Ethanol Injection (PEI) were performed. Overall, 61 procedures met the criteria for plasma or Plt infusion and
major bleeding complications occurred in 3 patients (0,6%). In 2
patients, anemia-related paracentesis was treated by blood transfusions whereas one patient following variceal band ligation had
to be hospitalized for severe anemia. Major complications rate was
1.6% in Plt/plasma infusion exposed patients versus 0.47% in unexposed (p = 0.28). Minor events with a > 1.5 g/dl haemoglobin decline
occurred in 17 patients (15 paracentesis and 2 band ligations), rate
was 3.2% in Plt/plasma infusion exposed patients versus 3.5% in
unexposed (p = 0.9).
Conclusion: Pre-treatment platelet transfusion in cirrhotic
patients with Plt count < 50,000/mmc and plasma infusion for
INR > 1.5, was associated with a negligible risk of bleeding and
appeared as a safe, cost/effective strategy.
L. Belloni 1 , G.A. Palumbo 2 , L. Li 3 , S.R. Chirapu 4 ,
L. Calvo 1,2 , L. Lupacchini 1 , M.G. Finn 4 ,
U. Lopatin 5 , A. Zlotnick 3,5 , M. Levrero 2
1 Centre for Life Nanoscience, IIT-Sapienza, Rome,
Italy
2 Department of Internal Medicine – DMISM,
Sapienza University of Rome, Italy
3 Department of Molecular & Cellular Biochemistry,
Indiana University, Bloomington, USA
4 Department of Chemistry, Georgia Institute of
Technology, Atlanta, USA
5 Assembly Bioscience, Bloomington, USA
Introduction and aim: HBV core protein (Cp) represents an
attractive new therapeutic target for HBV chronic infection. Cp
has been shown to bind the nuclear cccDNA mini-chromosome as
well as a number of cellular genes promoters. Several compounds
that target Cp and HBV capsids assembly, including the heteroaryl-dihydropyrimidines (HAPs) and the phenyl-propenamide
derivatives AT61 and AT130, have been shown to inhibit HBV replication in vitro and in vivo. HAPs and AT130 enhance the rate of Cp
assembly and stabilize preferentially non-capsid polymers of Cp.
Here we investigated the ability of the Core protein Assembly Modulators (CaMPs) HAP12 and AT130 to affect both nuclear cccDNA
transcription and cytoplasmic capsid assembly Cp functions.
Methods: HAP12 and AT130 effects on capsid-associated HBVDNA, cccDNA and pgRNA levels (quantitative real-time PCR with
specific primers) were assessed in: (a) HBV-infected NTCP-HepG2
cells; (b) AD38 inducible HBV stable cell line. Recruitment of HBc
and histone modifications on the viral minichromosome were
assessed using the cccDNA ChIP assay in AD38 cells.
Results: CaMPs treatments resulted in a very strong inhibition of
HBV replication (>95%) and a significant but incomplete reduction
of the stable cccDNA pool. A strong effect on cccDNA-dependent
HBeAg production (AD38 tet-off) and pgRNA transcription (AD38
tet-off/tet-on and NTCP-HepG2 infected cells) was also demonstrated. The ability of HAP12 to target cccDNA transcription was
confirmed by the reduced cccDNA-bound H3 histone acetylation
and the decreased HBc occupancy on the cccDNA in induced AD38
cells. Importantly, when CaMPs treatment was started during infection, cccDNA formation/accumulation was completely inhibited
(>95%) and viral replication was blunted.
Conclusions: Anti-capsid compounds (CpAMs) have an impact
on Cp nuclear functions at multiple levels: block of new cccDNA
formation/accumulation, reduction of an established cccDNA pool
and inhibition of HBc occupancy and histone acetylation on the
cccDNA that translate into a reduced pgRNA transcription.
OC-21
OC-22
RESIDUAL HCV-RNA IN LIVER EXPLANTS FROM
PATIENTS UNDERGOING SOFOSBUVIR AND
RIBAVIRIN TREATMENT WHILE AWAITING LIVER
TRANSPLANTATION IS NOT ASSOCIATED WITH
HCV RELAPSE
A HYPER-GLYCOSYLATION OF HBV SURFACE
MAJOR HYDROPHILIC REGION CORRELATES
WITH IMMUNOSUPPRESSION-DRIVEN HBV
REACTIVATION AND HAMPERS HBSAG
RECOGNITION IN VITRO
M. Gambato 1,2 , S. Pérez-del-Pulgar 1 ,
C. Hedskog 3 , J. Svarovskia 3 , M.S. Paulson 3 ,
J. Denning 3 , M.P. Curry 4 , N. Caro-Pérez 1 ,
M.C. Londoño 1 , X. Forns 1
L. Colagrossi 1 , R. Salpini 1 , M. Surdo 1 , A. Battisti 1 ,
M. Pollicita 1 , A. Bertoli 1 , F. Di Santo 1 , C. Becker 2 ,
C. Mastroianni 3 , M. Marignani 4 , S. Maylin 5 ,
C. Delaugerre 5 , F. Morisco 6 , N. Coppola 7 ,
A. Marrone 8 , L. Sarmati 9 , M. Andreoni 9 ,
M. Angelico 10 , J. Verheyen 11 , C.F. Perno 1 ,
V. Svicher 1
1 Liver Unit, Hospital Clinic, IDIBAPS, CIBERehd,
Barcelona, Spain
2 Multivisceral Transplant Unit, Department of
Surgery, Oncology and Gastroenterology, Padua
University Hospital, Padua, Italy
3 Gilead Sciences Inc., Foster City, CA, USA
4 Beth Israel Deaconess Medical Center, Boston, MA,
USA
Background and aims: Sofosbuvir (SovaldiTM , SOF) in combination with ribavirin (RBV) administered to HCV-infected patients
with hepatocellular carcinoma awaiting liver transplant (LT) prevented recurrence of HCV post-LT in 70% in those individuals with
undetectable serum HCV-RNA at time of LT (Curry et al., Gastroenterology, 2014). The present study examined the presence of
HCV-RNA in available liver explants from patients in the Phase 2
clinical study.
Patients and methods: Liver explant samples were collected
from 38 HCV-infected cirrhotic patients that underwent LT. All
patients received 3–52 weeks of SOF + RBV while awaiting LT. HCVRNA levels in the liver were determined by quantitative real-time
PCR in triplicate experiments. Liver explants from 39 HCV-RNA
negative patients were used as controls.
Results: Thirty-four patients were included in the final analysis.
Twenty-three out of 34 (68%) patient liver explants were HCV-RNA
positive (range 0.7–67566 copies/␮g) and 11 (32%) were HCV-RNA
negative. HCV-RNA was not detected in any of the control explants.
Treatment duration and time with undetectable HCV-RNA in serum
before LT were significantly lower in patients with HCV-RNA in
liver explants (16 and 7 weeks, respectively), compared to those
with undetectable HCV-RNA (24 and 13 weeks, p = 0.037, p = 0.045,
respectively). Twenty-four out of 34 patients (71%) achieved SVR12
after LT and 10 patients (29%) presented recurrent HCV infection.
Interestingly, HCV-RNA was detected in liver explants from 16
(68%) of the 24 responders and in 7 (70%) of the 10 non-responders
(p = 0.850).
Conclusions: SOF + RBV regimen is an efficacious therapy for
preventing HCV recurrence. The presence of HCV-RNA in liver
explants did not significantly correlate with the likelihood of recurrent HCV infection post-LT. Residual HCV in liver explants may
represent non-functional HCV-RNA (incomplete genomes versus
unfit viral strains) in the context of HCV-RNA polymerase inhibition
by SOF.
e11
1
Department of Experimental Medicine and Surgery,
Tor Vergata University, Rome, Italy
2 Institute of Virology, University of Cologne,
Germany
3 “Sapienza” University, Rome, Italy
4 Department of Gastroenterology, S.Andrea
Hospital, Rome, Italy
5 Laboratoire de Virologie, AP-HP Hopital
Saint-Louis, Paris, France
6 Department of Clinical Medicine and Surgery,
University of Naples Federico II, Section of Infectious
Diseases, Naples, Italy
7 Department of Mental Health and Public Medicine,
Section of Infectious Diseases, Second University of
Naples, Naples, Italy
8 Internal Medicine and Hepatology Unit, Second
University of Naples, Naples, Italy
9 Infectious Diseases Unit, Tor Vergata University
10 Hepatology Unit, Tor Vergata University Hospital,
Rome, Italy
11 Robert Koch Institute of Virology, University of
Essen, Essen, Germany
Introduction/aim: To investigate N-linked glycosylation patterns of HBsAg in immunosuppression-driven HBV-reactivation
and to evaluate their impact on HBsAg-antigenicity.
Methods: Mutations associated with acquisition of Nglycosylation site were investigated in 127 HBsAg genotype-D
sequences from 47 patients with immunosuppression-driven
HBV-reactivation (defined as Hwang, 2014), and 80 chronically
HBV-infected drug-naïve patients as control.
The impact of N-glycosylation sites on HBsAg-antigenicity
was analyzed by transfecting HepG2-cells with a plasmid encoding wild-type and mutated HBsAg linked to a streptavidin-tag
(strep-tag). The strep-tagged HBsAg amount in supernatants was
quantified by a specifically designed ELISA targeting the Strep-tag
(thus, not affected by HBsAg-mutations), and by ELISAs targeting
the HBsAg (Architect-Abbott, Monolisa-Biorad).
Results: Additional N-glycosylation sites are found in 19.1%
(9/47) of HBV-reactivated patients versus 0/80 controls (P < 0.001).
They localize in the major hydrophilic HBsAg-region (MHR), target of antibodies. In 7 patients, a single additional N-glycosylation
site results from the mutations T115N (n = 2), T123N (n = 2),
T131N (n = 2), and from the insertion of an N between 114 and
115 position (ins114–115N) (n = 1). In the remaining 2 patients,
2 additional glycosylation sites result from S113N+T131N and
ins114–115N+T117N, respectively.
Notably, 5/9 patients with ≥1 additional N-glycosylation sites
remain HBsAg-negative by diagnostic-test at HBV-reactivation
(P = 0.002).
e12
In vitro, all additional N-glycosylation sites decrease the
strep-tagged HBsAg quantification by the 2 ELISAs targeting the
HBsAg. In particular, T115N, T123N, ins114–115N determine a
>90% decrease in HBsAg-quantification by both ELISAs. Similarly,
ins114–115N+T117N cause a 90.2% and 75.4% reduction in HBsAgquantification, respectively. No decrease of strep-tagged HBsAg
is revealed by ELISA targeting the Strep-tag. This suggests that
additional N-glycosylation sites hamper HBsAg-recognition by
antibodies without affecting HBsAg-release.
Conclusions: Additional N-glycosylation sites in MHR correlate
with false HBsAg-negativity at ELISA despite HBV-reactivation, and
profoundly affect HBsAg-antigenicity in vitro. This supports the role
of immune-escape mutations in HBV-reactivation during immunesuppression and the importance of HBV-DNA (more than HBsAg)
in HBV-reactivation diagnosis.
OC-23
HBSAG LOSS IS ENOUGH TO DISCONTINUE
LONG-TERM NUCLEOS(T)IDE ANALOGUE
THERAPY IN HBEAG-NEGATIVE CHRONIC
HEPATITIS B PATIENTS IN REAL PRACTICE?
M. Fasano 1 , M. Ciarallo 1 , G. Niro 2 , R. Fontana 2 ,
R. Cozzolongo 3 , A. Maci 4 , I. Carraturo 4 ,
A. Miglietta 4 , G. Angarano 5 , T. Santantonio 1
1
Infectious Diseases, University of Foggia, Italy
Gastroenterology, Casa Sollievo Sofferenza
Hospital, San Giovanni Rotondo (FG), Italy
3 Division of Gastroenterology 1, National Institute of
Gastroenterology S. de Bellis, Castellana Grotte, Bari,
Italy
4 Infectious Diseases, Vito Fazzi Hospital, Lecce, Italy
5 Infectious Diseases, University of Bari, Italy
2
Introduction: Currently, oral nucleos(t)ide analogues (NAs) are
the most common therapeutic strategy for treatment of chronic
hepatitis B (CHB). HBsAg loss is a well-established endpoint of
therapy, but it is still unclear whether HBsAg clearance alone is
enough to stop long-term NA therapy or seroconversion to anti-HBs
is required.
Aim: To evaluate in clinical practice the outcome in HBeAgnegative CHB patients who discontinued NA therapy after HBsAg
loss.
Methods: HBeAg-negative CHB patients with HBsAg clearance
during long-term NA therapy followed for at least 24 weeks after NA
discontinuation were retrospectively examined. HBsAg, anti-HBs,
HBV DNA levels and alanine aminotransferase levels were monitored every 1–3 months after NA discontinuation in the first year
and every 6 months thereafter. Virological, biochemical and clinical
data were collected and entered into a database.
Results: Among 590 monoinfected HBeAg-negative CHB
patients who received NA therapy between 1998 and 2009, 24
(4%) lost HBsAg after a median duration of therapy of 103 months
(range 22–180) and were included in this study. Median age was 53
years (range 34–170), 18 were males, 18% cirrhotics. Twenty-two
patients received NA monotherapy (15 lamivudine, 3 entecavir, 4
tenofovir) and two patients received lamivudine + adefovir combination therapy. Only 11/24 patients (46%) developed anti-HBs
(anti-HBs level: ≤100 IU/ml in 5 and 100–500 IU/ml in 6), whereas
13 patients remained persistently anti-HBs negative during followup. All 24 patients stopped NA treatment after a median duration
of consolidation therapy of 12 months (range 0–70). No case of
virological and biochemical breakthrough were reported during 24
months (range 6–180) of post-treatment follow-up.
Conclusion: HBsAg loss is a rare event in HBeAg-negative CHB
patients treated with NAs. However, regardless of anti-HBs seroconversion and duration of consolidation therapy, long-term NA
therapy can be safely discontinued in clinical practice in patients
whit HBsAg loss, even in those with advanced liver disease.
OC-24
HIGH CAPABILITY OF CONTRAST ENHANCED
ULTRASOUND IN DEFINING A RAPID DIAGNOSTIC
AND THERAPEUTIC WORK-UP FOR NODULES
<2 CM IN CIRRHOTICS DURING SURVEILLANCE
A. Giorgio 1 , G. Iaquinto 2 , L. Montesarchio 1 ,
P. Gatti 3 , B. Santoro 4 , F. Amendola 5 ,
P. Matteucci 6 , C. Coppola 2,7 , V. Giorgio 7,8
1 Interventional Ultrasound Unit, Tortorella Clinical
Institute, Consorzio ISMESS, Salerno, Italy
2 Unità Operativa di Medicina Interna, Casa di Cura
S. Rita, Atripalda, Avellino, Italy
3 Unità Operativa di Medicina Interna, Ospedale di
Fasano, Brindisi, Italy
4 Unità operativa ecografia interventistica, Athena,
Caserta, Italy
5 Unità Operativa di Medicina Interna, Tortorella
Clinical Institute, Consorzio ISMESS, Salerno, Italy
6 Campus biomedico, Roma, Italy
7 Unità Operativa di Medicina Interna, Epatologia
interventistica, ospedale di Gragnano, Napoli, Italy
8 Università Cattolica del Sacro Cuore, Policlinico
Gemelli, Roma, Italy
Introduction: The disappearance of portal blood flow and the
arterial vascularization is the hallmark of hepatocarcinogenesis,
leading from benign regenerating nodules to well differentiated
HCC. The capability of a dynamic imaging modality detecting arterial hypervascurarization of small nodules is crucial to promote a
rapid diagnostic and therapeutic work-up improving survival.
Aim: To evaluate the capability of CEUS to detect arterial vascularization of ≤2 cm HCC nodules arising during surveillance
shorting diagnostic and therapeutic work-up.
Materials and methods: From July 2011 to June 2014 among
1757 consecutive cirrhotics under surveillance with Ultrasound
(US), 229 new single nodules 7–20 mm in diameter were detected.
All these patients underwent CEUS followed by US guided percutaneous 18 G needle core biopsy of the nodules as gold standard. Of
the 229 nodules, 27 were hyperechoic lesions, 171 hypoechoic and
31 isoechoic.
Results: On histology, 199/229 nodules were HCC and 30 were
benign. Of the 199 HCCs, CEUS showed arterial hypervascularity
in 190 nodules (95.5%) [sensitivity 94.48%; specificity 100%; PPV
100%, NPV 76.92%]. Of the 39 CEUS arterial-unenhanced nodules,
30 were benign and 9 (4,5%) were well-differentiated HCC.
The fate of the 9 arterial-unenhanced CEUS HCC patients was
also compared with that of the 6 HCC patients with no arterial
enhancement on ultrafast dynamic gadolinium-enhanced MRI performed for staging in all HCC patients included in the study. All
these 15 patients were treated with percutaneous radiofrequency
ablation: 1-year distant recurrence rate was 11.1% in CEUS group
and 30% in MRI group (p < 0.01).
Conclusions: CEUS has a great capability to detect arterial
hypervascularity of small HCC. Because only 4.5% of new nodules
escape the demonstration of arterial hypervascularity, CEUS must
be performed immediately after conventional US to contrast the
malignant fate of small lesions arising in cirrhotics.
OC-25
MIR-17/92 EXPRESSION PATTERN: A
MOLECULAR SIGNATURE OF HCV-RELATED
MIXED CRYOGLOBULINEMIA
A. Piluso ∗ , L. Gragnani, A. Genovesi, E. Fognani,
M. Monti, T. Urraro, A.L. Zignego
Department of Experimental and Clinical Medicine,
University of Florence, Florence, Italy
Introduction: HCV infection is closely related to the development of lymphoproliferative disorders (LPDs), mainly mixed
cryoglobulinemia (MC) and B-cell lymphoma. Modification of the
expression levels of specific microRNAs (miRNAs) has been associated with different autoimmune and/or LPDs. In particular, the
endogenous miR-17/92 cluster is very often amplified in cancer
and in autoimmunity. Scarce data exist about the modifications of
miRNA expression levels in HCV-related LPDs.
Aim: To evaluate the modifications of miR-17/92 cluster levels
in HCV-positive patients with and without MC.
Methods: miR-17/92 cluster expression levels were evaluated
by Real Time PCR in PBMC samples from 79 HCV patients: 34
without LPD [HCV] and 45 with MC [HCV-MC]; among the 45
MC patients were included 20 patients who experienced a sustained virological and clinical response after antiviral treatment
and of which pre and post therapy sampling was available. Relative expression levels of all the members of the miR-17/92 cluster
(namely, miR-17, miR-18a, miR-19a, miR-19b, miR-20a and miR92a) were evaluated with the 2−Ct method, using miR-let-7d
as housekeeping.
Results: All the members of the miR-17/92 cluster were highly
upregulated in PBMCs from HCV-MC patients (p < 0.001) when
compared to HCV group. A restoration of miRNAs levels was
observed in the samples taken after viral eradication (miR-17, miR18a, miR-19a, miR-19b, miR-20a, p < 0.001 and miR-92a, p < 0.05,
when compared with pre-treatment levels). Regarding miR-20a,
the levels in samples taken after HCV eradication continued to be
significantly higher than in controls (HCV patients) (p < 0.05), in
spite of the sudden fall observed after therapy.
Conclusions: This study shows that the pattern of miRNA-17/92
cluster is modified in PBMC from HCV patients with MC. The sudden
restoration of these values of expression in patients achieving a
sustained virological and clinical response after antiviral treatment,
strongly suggests that miR-17/92 cluster plays a key role in the
pathogenesis of MC.
e13
OC-26
THE ITALIAN ENTAS COHORT STUDY: ENTECAVIR
EFFECTIVENESS IN NAïVE AND TREATMENT
EXPERIENCED PATIENTS WITH CHRONIC
HEPATITIS B
E. Porro 1 , A. Di Leo 2 , A. Marzano 3 ,
G. Brancaccio 4 , S. Maimone 5 , M. Fasano 6 ,
A. Grasso 7 , F. Bronte 8 , S. Fagiuoli 9 ,
T. Santantonio 6 , F. Morisco 10 , E. Petrelli 11 ,
G. Surace 12 , G. Labbadia 13 , L. Badia 14 ,
G.A. Niro 15 , M. Vinci 16 , A. Montineri 17 ,
F. Vinelli 18 , M. Massari 19 , L. Nosotti 20 ,
G. Galati 21 , G. Missale 1,22 , Entas Study Group
1
Infectious Diseases and Hepatology, Azienda
Ospedaliero-Universitaria di Parma, Parma, Italy
2 Gastroenterology and Digestive Endoscopy,
University of Bari, Bari, Italy
3 Gastroepatologia, Ospedale San Giovanni Battista,
Torino, Italy
4 Epatiti Virali Acute e Croniche, II Università degli
Studi di Napoli, Napoli, Italy
5 Clinical and Molecular Hepatology, University
Hospital of Messina, Messina, Italy
6 Malattie Infettive, Università degli Studi di Foggia,
Foggia, Italy
7 Gastroenterologia Osp San Paolo, Savona, Italy
8 Gastroenterologia, Di.Bi.M.I.S., University of
9 Gastroenterologia ed Epatologia Dei Trapianti,
Ospedali Riuniti di Bergamo, Bergamo, Italy
10 Gastroenterology Unit, Department of Clinical
Medicine and Surgery, University of Naples, Federico
II, Napoli, Italy
11 Malattie Infettive, Azienda Ospedaliera San
Salvatore, Pesaro, Italy
12 Clinica di Gastroenterologia, Università
Politecnica delle Marche, Ancona, Italy
13 Dipartimento di Medicina Interna e Specialità
Mediche, UOC Medicina Interna F, Sapienza,
Università di Roma, Umberto I Policlinico, Roma,
Italy
14 Istituto Malattie Infettive, Azienda Ospedaliero
Universitaria Policlinico S.Orsola Malpighi, Bologna,
Italy
15 Gastroenterology, Casa Sollievo Sofferenza
Hospital, IRCCS, San Giovanni Rotondo, Italy
16 SC Epatologia e Gastroenterologia AO Ospedale
Niguarda Cà Granda, Milano, Italy
17 Malattie Infettive, PO Ferrarotto, Catania, Italy
18 Gastroenterologia ed Endoscopia Digestiva,
Azienda Ospedaliero Universitaria di Foggia, Foggia,
Italy
19 Infectious Diseases, IRCCS, Azienda Ospedaliera S.
Maria Nuova, Reggio Emilia, Italy
20 National Institute for Health, Migration and
Poverty (NIHMP), Italy
21 Unità di Medicina Clinica-Epatologia, Policlinico
Universitario Campus Bio Medico, Italy
22 AISF Associazione Italiana per lo Studio del Fegato,
Roma, Italy
Introduction: Real-life studies, mainly report results in
untreated patients.
e14
Aims: To define Entecavir antiviral potency and efficacy in
untreated and IFN- and NUC-experienced patients in clinical
practice.
Material and methods results: 423 patients were evaluated for
undetectable HBV-DNA, HBeAg seroconversion and monitored for
liver function and main complications.
304 patients were untreated (naive), 97 IFN- and 34 NUCexperienced (12 IFN-experienced). Forty-five percent had liver
cirrhosis. Rate of undetectable HBV-DNA was 85–90% at 24–30
months in the whole cohort, while 84–90%, 87–91% and 85–80%
in naive, IFN- and NUC-experienced. Nine patients had been previously treated with LAM or ADV and were not on treatment while 25
switched to ETV or added ETV at enrollment. Two patients switched
to TDF, one because of virologic failure (L180M, M204V, M250V)
and the second for persistence of low viremia (<3 log 10) a third subject added TDF because of partial virologic response. Two patients
are still viremic, one not compliant and the second is on dialysis
treatment.
Thirty of 56 HBeAg positive patients seroconverted to anti-HBe
(53.5%) and 27 lost HBsAg (6.5%).
Bilirubin > 2 mg/ml, albumin < 3.5 g/dl and INR > 1.7 was in 15%,
22% and 8% of cases at baseline with significant improvement at
24 months (bilirubin, p < 0.001; albumin and INR, p < 0.05). Ascites
resolved in 22 of 37 cases while developed in 14 cases; 6 variceal
bleedings were observed. HCC was present in 41 patients at baseline and 30 developed HCC during follow-up with an incidence of
2.5% per year; 80% of new HCC cases were cirrhotic and significantly
older (p < 0.0001), while viremia, HBeAg status, coinfection (HCV,
HDV), ALT levels and BMI were not statistically different compared
to the other patients.
Conclusions: The study of Entecavir treatment in field-practice
confirms its efficacy in suppressing HBV replication also in
treatment-experienced patients. Analysis of HCC rate confirms
results of other real-life studies.
OC-27
NOTCH4 AND MHC CLASS II POLYMORPHISMS
CONTRIBUTE TO HCV-RELATED BENIGN AND
MALIGNANT LYMPHOPROLIFERATIVE DISEASES
A. Piluso 1,∗ , L. Gragnani 1 , A. Genovesi 1 ,
V. De Re 2 , E. Fognani 1 , M. Libra 3 , A.L. Zignego 1
1 Department of Experimental and Clinical Medicine,
2 Bio-Proteomics Facility, Department of
Translational Research, CRO, National Cancer
Institute, Aviano, Italy
3 Department of Biomedical Sciences, Section of
Pathology and Oncology, Laboratory of Translational
Oncology and Functional Genomics, University of
Catania, Catania, Italy
Introduction: Hepatitis C virus (HCV) can cause lymphoproliferative disorders (LPDs). The most frequent one is mixed
cryoglobulinemia (MC), clinically benign, but evolving in about
10% of cases into a non-Hodgkin’s lymphoma (NHL). Recently, a
GWAS allowed the discovery of an association of two SNPs on chromosome 6 and HCV-related MC; the first one is an intronic SNP
(rs2071286) of the NOTCH4 gene, the second one (rs9461776) is
located between HLA-DRB1 and HLA-DQA1 gene segments.Aim:
To define the contribution of rs2071286 and rs9461776 in the predisposition to develop HCV-related LPDs.
Methods: rs2071286 and rs9461776 were determined in 438
patients: 85 HCV patients without LPDs (HCV), 73 HCV patients
with circulating cryoglobulins (MC-HCV), 108 with HCV-associated
MC syndrome (MCS-HCV), 62 with HCV-related NHL (NHL-HCV)
and 110 HCV-negative patients with NHL (NHL).
Results: Concerning rs2071286, significantly higher minor
allele frequency (maf) was observed in all the cases respect to controls (MC-HCV, p = 0.035; MCS-HCV, p < 0.001; NHL-HCV, p = 0.001
and NHL, p = 0.019). Comparing to HCV group, the odds-ratio associated with rs2071286 maf were: MC-HCV, OR 1.79; MCS-HCV, OR
2.59; NHL-HCV, OR 2.6; NHL, OR 1.81.
Regarding rs9461776, the higher maf was observed in NHL-HCV
and in NHL groups respect to HCV (p = 0.011 and p = 0.0283, respectively). The presence of minor allele of rs9461776 conferred an OR
of 2.42 to NHL-HCV and an OR of 1.7 to NHL group, in respect to
controls.
Conclusions: We confirm the previously demonstrated association between the two SNPs and HCV-related MC vasculitis, and
showed similar associations for HCV-related NHLs and, regarding
rs2071286, also for patients with CGs but without symptoms. Furthermore, HCV-negative NHLs showed higher frequencies of the
two minor alleles respect to controls, but lower compared to HCVpositive lymphomas. This suggests that HCV acts on a permissive
genetic substrate and confirms the virus contribution to the lymphomagenesis.
OC-28
17␤-ESTRADIOL INHIBITS HEPATITIS C (HCV)
INFECTION VIA BINDING TO ITS RECEPTOR
P. Giarda, A. Magri, C.Z. Foglia, E. Boccato,
E.M. Burlone, R. Minisini, E. Grossini, M. Pirisi
Department of Translational Medicine, Univertità
del Piemonte Orientale, Novara, Italy
Background: Hormonal factors may play a role in controlling hepatitis C virus infection (HCV). In fact, spontaneous HCV
clearance is more common among premenopausal women than in
men and, when infection persists, histologic progression of chronic
hepatitis is slower in pre-menopausal women in comparison to
postmenopausal women and men. Aim of our study has been to
evaluate sex hormones as inhibitors of HCV replication.
Methods: Huh-7.5 cells infected with the JFH-1 virus were
exposed to one each among the following hormones: dehydroepiandrosterone (DHEA), testosterone, progesterone and 17␤estradiol (the latter in the presence/absence of the estradiol
receptor antagonist fulvestrant). Based on the inhibition of viral
replication, dose–response curves covering the physiological range
were established allowing calculation of IC50 values. In model A,
hormones were added 3 h post-infection. In model B, Huh-7.5 cells
were incubated with hormones for 1 h before being infected; 3 h
later, the inoculum was replaced with fresh medium. In model C,
a 16 h pre-incubation 17␤-estradiol preceded replacement with
fresh medium, followed by infection.
Results: Progesterone and testosterone showed no inhibitory
effect on viral replication at the concentrations tested for both
model A or B. DHEA exhibited only a partial antiviral effect in both
model (39%). In contrast, in model A 17␤-estradiol inhibited viral
replication up to 45%, allowing to estimate IC50 = 490 nM. Moreover,
the inhibitory effect of 17␤-estradiol reached 67% in model B and
64% in model C, with IC50 values = 140 and 160 nM, respectively.
Although fulvestrant did not exhibit any appreciable effect on viral
replication, in its presence the inhibition exerted by 17␤-estradiol
was reverted in a dose-dependent manner.
Conclusions: In vitro, 17␤-estradiol is able to block HCV infection, likely by modulation of intracellular pathways following
binding to the estradiol receptor, whose activation leads to an
antiviral state.
OC-29
ACQUIRED SPHEROCYTIC LIKE ANEMIA
COMBINED WITH INEFFECTIVE
ERYTHROPOIESIS SUSTAINS ANEMIA IN
PATIENTS WITH CHRONIC HEPATITIS C
INFECTION RECEIVING TELAPREVIR OR
BOCEPREVIR-BASED TRIPLE THERAPY
Lupo 1 ,
Russo 2,3 ,
Iolascon 2,3 ,
Ieluzzi 4 ,
F.
R.
A.
D.
P. Toniutto 5 , S. Piovesan 6,7 , E. Raffetti 8 ,
A. Siciliano 1 , A. Matte’ 1 , F. Turrini 9 , F. Donato 8 ,
A. Alberti 6 , V. Zuliani 1 , G. Fattovich 1,4 ,
L. De Franceschi 1
1 Department of Medicine, University of Verona,
Verona, Italy
2 Dipartimento di Medicina Molecolare e
Biotecnologie Mediche, Università degli Studi di
Napoli Federico II, Napoli, Italy
3 CEINGE Biotecnologie Avanzate, Napoli, Italy
4 Clinical Unit of Gastroenterology, Azienda
Ospedaliera Universitaria Integrata Verona, Verona,
Italy
5 Department of Medicine and Pathology Clinical and
Experimental, Medical Liver Transplantation Unit,
Internal Medicine, University of Udine, Udine, Italy
6 Department of Molecular Medicine, University of
Padua, Padua, Italy
7 Clinical Unit of General Medicine, Azienda
Ospedaliera Universitaria Integrata Padua, Padua,
Italy
8 Institute of Hygiene, Epidemiology and Public
Health, University of Brescia, Brescia, Italy
9 Department of Oncology, University of Turin, Turin,
Italy
Background and aims: The addition of protease inhibitors,
boceprevir (BOC) or telaprevir (TVR), to peg-interferon and ribavirin (PR) increases the incidence of anemia. Although genetic
variants in ITPA gene have been linked to the hemolytic anemia
induced by PR, the mechanism sustaining the severe anemia during
triple therapy is still unknown.
Materials and methods: We studied 59 patients with chronic
hepatitis C: 29 treated with TVR/PR and 30 with BOC/PR. Patients
were examined at baseline, at week 4 (end of PR lead-in phase),
at 12, 16 and 24 weeks of treatment. In all patients we evaluated
biochemical and hematological parameters, red cell index; while
in a subgroup, we carried out in vitro functional studies to dissect the mechanism(s) underlying anemia in triple therapy. IL28B
(rs12979860) and ITPA (rs1127354, rs7270101) polymorphisms
were also analyzed.
Results: In chronic HCV patients on triple therapy, we found
an early acute normochromic normocytic hemolytic anemia (4–8
weeks) followed by a late macrocytic hypo-regenerative anemia with low reticulocyte count in response to the degree of
anemia (12–24 weeks). Although the beneficial effects of ITPA polymorphisms impacted the early phase of anemia (4 weeks), the
e15
functional studies on red cells revealed: (i) the presence of spherocytes; (ii) increased osmotic fragility; (iii) changes in red cell
membrane protein composition; (iv) increased phosphorylation
of ␤-adducin with reduced membrane-cytoskeleton stability; (v)
increased release of erythroidmicroparticles.
Conclusions: Our data indicate that the bimodal pattern of anemia in chronic HCV patients on triple therapy might be induced by
acquired spherocytic-like anemia as dominating component in the
early phase (4–8 weeks), followed by the appearance of hyporegenerative macrocytic anemia (12–24 weeks), most likely related
to the combination effects of PR and TVR or BOC on erythropoiesis.
These data could guide strategy of anemia management during
triple therapy.
OC-30
SOFOSBUVIR-BASED ALL-ORAL TREATMENT FOR
ELDERLY CHRONIC HEPATITIS C PATIENTS: A
COST-EFFECTIVENESS ANALYSIS
A. Ciaccio 1 , P.A. Cortesi 2 , G. Bellelli 3 , M. Rota 4 ,
S. Okolicsanyi 1 , M. Rota 1 , L. Mantovani 2 ,
G. Annoni 3 , M. Strazzabosco 1,5
1 Department of Surgery and Translational Medicine,
University of Milano-Bicocca, Milan, Italy
2 Research Centre on Public Health (CESP), University
of Milano-Bicocca, Milan, Italy
3 Department of Health Sciences, Geriatric Medicine,
4 Department of Health Sciences, Centre of
Biostatistics for Clinical Epidemiology, University of
Milan-Bicocca, Milan, Italy
5 Liver Center & Section of Digestive Diseases,
Department of Internal Medicine, Yale University
School of Medicine, New Haven, CT, United States
Background and aim: A relevant proportion of patients affected
by chronic hepatitis C (CHC) is older than 65 years. Comorbidities and a higher susceptibility to drugs toxicity have historically
limited treatment in these patients. Recent approval of interferonfree regimens, characterized by high efficacy and limited toxicity,
provides unprecedented chances for these patients to be cured.
The aim of this study is to assess cost-effectiveness, taking into
account the severity of liver disease, age, and the geriatric (frailty)
status.
Methods: A semi-Markov model of CHC natural history was
built. The study focuses on CHC patients older than 65 years, stratified according to liver fibrosis (METAVIR F3 and F4), age (65–85
years old) and Fried’s frailty phenotype (not frail, pre-frail and frail)
generating 30 simulated cohorts. Treatment with sofosbuvir plus
simeprevir (SOF/SMV) versus no treatment was assessed for each
cohort. The model estimated costs, Life Years and Quality Adjusted
Life Years (QALY), with a lifetime time horizon and the Health
System perspective. Results are presented as incremental costeffectiveness ratios (ICERs) per QALY gained. Cost-effectiveness
was defined as an ICER under the 37,000D threshold.
Results: At each fibrosis score, ICER increased with age and
frailty index. Among F3 patients, ICER ranged from D13,934 in notfrail 65-years-old and D79,354 in frail 85-years-old patients. Among
F4 patients ICER ranged from D13,873 in not frail 65-years-old and
D115,965 in frail 85-years-old patients. In both F3 and F4 cohorts
ICER was below D37,000/QALY up to age 80 in non-frail patients,
up to age 75 in pre-frail patients, up to age 70 in frail patients.
e16
Conclusion: SOF/SMV treatment is cost-effective in most CHC
patients older than 65 years, however a careful assessment of the
patient geriatric status is mandatory. This cost-effectiveness analysis should promote a prospective clinical study to verify efficacy
and side effects in elderly HCV patients.
long natural history of the disease, these OIs have proven to be easy
to collect and to work appropriately. Therefore, they are suitable to
be extended to specialized centres involved in PSC management to
further validate their clinical usefulness.
OC-31
OUTCOME INDICATORS IN PRIMARY
SCLEROSING CHOLANGITIS: INTERIM ANALYSIS
OF THE VALUE-BASED MEDICINE IN
HEPATOLOGY STUDY
L. Fabris 1,2 , A. Ciaccio 3 , S. Okolicsanyi 3 , M. Rota 4 ,
P.A. Cortesi 5 , M.R. Buonocore 3 , M. Gemma 3 ,
P. Giani 6 , L.S. Belli 7 , S. Fagiuoli 6 , L. Scalone 5 ,
M.G. Valsecchi 4 , L.G. Mantovani 5 ,
M. Strazzabosco 2,3
Padua, Padua, Italy
Milano-Bicocca, Milan, Italy
of Milan-Bicocca, Milan, Italy
6 Department of Gastroenterology, Papa Giovanni
XXIII Hospital, Bergamo, Italy
7 Department of Hepatology and Gastroenterology,
Liver Unit, Niguarda Hospital, Milan, Italy
Introduction: Primary sclerosing cholangitis (PSC) is an enigmatic disease with scarce therapeutic options. The clinical
management of PSC remains challenging and may benefit from
Outcome Indicators (OI) to assess the quality of care.
Aims: This study aims to: (A) identify OIs for PSC, and (B) validate
OIs in a clinical context.
Methods: (A) A panel of experts generated a list of OIs by Delphi
method. (B) OIs with the highest RAND/UCLA score were tested in
an ongoing multicentric, prospective study (Value-based Medicine
in Hepatology, VBMH).
Results: Five OIs were identified having the highest rating values and low disagreement indexes: annual rate of acute cholangitis
episodes (OI#1); mortality rate for patients not yet listed for liver
transplantation (OI#2); rate of quality of life improvement, measured by EQ-VAS and EQ-5D (OI#3); number of patients died for
cholangiocarcinoma and colo-rectal carcinoma (OI#4); incidence
and/or worsening of osteoporosis (OI#5). In the validation study,
63 consecutive patients with PSC enrolled in 3 tertiary centres in
Northern Italy were evaluated for a median 24-months follow-up
period. For each OI, the following values were reported: (OI#1)
cumulative incidence of 5.2%, resulting in 0.029 cholangitis/patient;
(OI#2, OI#4) no patients died without being listed for transplantation or because of cancer during study time; (OI#3) 38.9 and 19.4%
of patients showed an improvement in EQ-VAS and EQ-5D parameters, respectively; (OI#5) 3% of patients developed or worsened
osteoporosis.
Conclusions: Five OIs for PSC were identified reporting high
consensus. Albeit the study population is small (as in the case of
rare diseases) and the follow-up time is short as compared to the
OC-32
HEPCIDIN RESISTANCE IN DYSMETABOLIC IRON
OVERLOAD
R. Rametta 1 , S. Pelusi 1 , P. Dongiovanni 2 ,
F. Iuculano 1 , A.L. Fracanzani 1,2 , S. Fargion 1,2 ,
L. Valenti 1,2
1 Department of Pathophysiology and
Transplantation, University of Milan, Milan, Italy
2 Department of Internal Medicine, Fondazione
IRCCS Cà Granda Ospedale Maggiore Policlinico,
Milan, Italy
Background and aim: The dysmetabolic Iron Overload Syndrome (DIOS) is defined by hyperferritinemia associated with
steatosis and insulin resistance, and is associated with increased
risk of hepatic and cardiovascular disease. The pathogenesis of iron
accumulation during DIOS is still unclear. Aim was to characterize
iron metabolism by performing an oral iron tolerance test (OITT)
in DIOS as compared to hereditary hemochromatosis (HH) and
healthy individuals.
Methods: In 17 healthy volunteers, 10 C282Y+/+ homozygous
HH, and 13 DIOS patients negative for mutations in HFE and FPN1,
we administered 65 mg ferrous-sulfate orally. All subjects had normal ferritin (30–150 ng/ml, >4weeks after depletion in HH and
DIOS), normal erythropoiesis and inflammatory indices. Ferritin,
transferrin saturation (TS%), and the iron hormone hepcidin were
evaluated at baseline (T0), 4 (T1), 8 (T2) and 24 (T3) h. Hepcidin was
measured by a new generation ELISA kit (DRG).
Results: HH had increased TS% and lower hepcidin than controls
and DIOS at all time points (p < 0.05). Despite baseline levels were
comparable to those of controls, DIOS patients had both higher
TS% and hepcidin levels at T1 and T3 (p < 0.05 for all). A hepcidin
release index did not differ between controls and DIOS, while it
was reduced in HH. Conversely, a hepcidin resistance index was
similar between HH and controls, while it was twofold higher in
DIOS compared to both controls and HH (p = 0.0004 and p = 0.004,
respectively).
Conclusions: Iron accumulation in DIOS is not secondary to
defective production of the hormone hepcidin, but it seems rather
due to resistance to the action of hepcidin in decreasing serum
iron and intestinal iron absorption. Additional studies are required
to clarify the underlying molecular mechanisms. Conversely, data
confirmed that inadequate hepcidin release underpins iron accumulation in HH. The present OITT protocol may represent a useful
instrument to study human iron metabolism.
OC-33
OC-34
ACUTE KIDNEY INJURY IN CIRRHOTIC PATIENTS
UNDERGOING CONTRAST-ENHANCED
COMPUTED TOMOGRAPHY
OBLITERATIVE PORTAL VENOPATHY IN THE
ABSENCE OF CLINICAL PORTAL HYPERTENSION:
AN UNEXPLORED PLANET
R. Filomia 1 , S. Maimone 1 , C. Saitta 1 , L. Visconti 2 ,
S. Caloggero 3 , A. Bottari 3 , A. Alibrandi 4 ,
I. Cacciola 1,5 , G. Caccamo 1 , S. Spinella 1 ,
D. Vadalà 1 , C.G. Gambino 6 , M.S. Franzè 6 ,
G. Gambino 6 , G. Raimondo 1,5 , G. Squadrito 1,5
M. Guido 1 , S. Sarcognato 1 , A. Sonzogni 2 ,
M.G. Lucà 3 , M. Senzolo 4 , S. Fagiuoli 3 ,
A. Ferrarese 4 , M. Pizzi 1 , L. Giacomelli 1 ,
G. Colloredo 5
1
Division of Clinical and Molecular Hepatology,
University Hospital of Messina, Messina, Italy
2 Division of Nephrology, University Hospital of
Messina, Messina, Italy
3 Department of Biomedical Sciences and
Morphological and Functional Imaging, University of
Messina, Messina, Italy
4 Department of Economical, Business and
Environmental Sciences and Quantitative Methods,
University of Messina, Messina, Italy
5 Department of Clinical and Experimental Medicine,
University Hospital of Messina, Messina, Italy
6 University School of Medicine of Messina, Messina,
Italy
Introduction: Contrast-induced acute kidney injury (CI-AKI)
following intravenous contrast medium administration is one of
the most common causes of hospital-acquired acute kidney damage.
Aim: To investigate the incidence and possible predisposing factors of CI-AKI in cirrhotic patients undergoing contrast-enhanced
computed tomography (CECT).
Patients and methods: We analysed 393 consecutively hospitalized cirrhotic patients. We excluded patients with active
bacterial infection, glomerular filtration rate (GFR) < 30 ml/min,
recently intake of antibiotics and anti-inflammatory drugs. CECT
was performed in 249 patients (CECT-group) and was not in the
remaining 144 (control group). No differences for age, sex, liver
disease aetiology, Child-Pugh (CP) and MELD scores were present
between the two groups. GFR was estimated using the Modification
of Diet in Renal Disease (MDRD-6) formula. The contrast inducednephropathy (CIN) and the AKI-network (AKIN) criteria were used
to assess CI-AKI development. Serum creatinine was evaluated just
before and 72 h after CECT examination and, analogously, it was
tested in control group at admission and 72 h later.
Results: CI-AKI occurred more frequently in CECT compared to
control group (22/249 vs 5/144, p < 0.05 according to AKIN criteria,
and 42/249 vs 14/144, p = 0.05 according to CIN criteria). The following variables were significantly associated with CI-AKI at the
multivariate regression analysis: male sex (p < 0.0001, OR: 0.118,
CI: 0.04–0.34), lower GFR (p = 0.022, OR: 1.019, CI: 1.003–1.035),
lower serum sodium levels (p = 0.025, OR: 0.87, CI: 0.770–0.982).
CI-AKI occurred independently of the presence of diabetes, arterial
hypertension, CP and MELD scores. Three-months follow-up was
available in 16/22 patients who developed CI-AKI showing that
serum creatinine persisted abnormally elevated in 10/16 (62.5%)
patients.
Conclusions: CECT is a risk factor for the development of CIAKI in hospitalized cirrhotic patients independently of the presence
of known causes of chronic kidney injury and of the severity of
liver disease. In these patients CECT-associated kidney dysfunction
seems to persist over time.
e17
1
Surgical Pathology & Cytopathology Unit,
Department of Medicine-DIMED, University of
Padua, Padua, Italy
2 Pathology Department, Papa Giovanni XXIII
Hospital, Bergamo, Italy
3 Gastroenterology and Transplant Hepatology, Papa
Giovanni XXIII Hospital, Bergamo, Italy
Surgical and Gastroenterological Sciences, University
Hospital of Padua, Padua, Italy
5 Department of Internal Medicine, San Pietro
Hospital, Ponte San Pietro (BG), Italy
Obliterative portal venopathy (OPV) is the primary histological
lesion associated with idiopathic non-cirrhotic portal hypertension (INCPH). Data on OPV in the absence of portal hypertension
(PH) are very scarce. This retrospective study aimed to assess the
prevalence of OPV lesions in patients with long-lasting abnormalities of liver function tests (LFTs) of unknown etiology, but
no evidence of PH. Four-hundred-eighty-two cases were retrospectively collected from the clinical and pathological databases
of three referral centers for liver diseases. Criteria for inclusion
involved the absence of clinical signs of PH and any known cause of
abnormal LFTs, and the histological demonstration of OPV lesions.
Histological findings were matched towards clinical and laboratory features. OPV lesions were identified in 19.9% of cases. LFTs
abnormalities were mild in most cases. Serum autoantibodies
turned positive in 54.2% of patients. Pro-thrombotic and autoimmune disorders were highlighted in 5.9% and 12.5% of tested
cases, respectively. Histologically, the most prevalent OPV-related
changes were aberrant vessels (96.9%) and portal angiomatosis
(74.0%). Among other INCPH-associated lesions, the most frequent were sinusoidal dilatation (65.6%) and increased number of
parenchymal veins (56.3%). Nodular regenerative hyperplasia was
detected in 9.6% of cases. Conclusions: OPV changes occur in a
consistent subgroup of patients with abnormal LFTs of unknown
origin, even in the absence of clinical PH. OPV lesions are frequently
associated with histological and clinical features commonly related
with INCPH, suggesting that patients may be in an early presymptomatic phase of INCPH. This condition is probably largely
underestimated throughout the clinical practice.
e18
OC-35
OC-36
ANTICOAGULATION DOES NOT INCREASE
PORTAL HYPERTENSION RELATED BLEEDING,
BUT EXPOSES PATIENTS WITH CIRRHOSIS TO A
HIGH RISK OF MINOR HEMORRHAGES: RESULTS
FROM A COMPARATIVE COHORT STUDY
DIAGNOSTIC ACCURACY OF LIVER AND SPLEEN
STIFFNESS MEASUREMENT FOR PORTAL
HYPERTENSION USING BIDIMENSIONAL SHEAR
WEAVE ELASTOGRAPHY
V. La Mura 1,2,∗ , S. Braham 3 , F. Branchi 4 ,
M. Moia 3 , A.L. Fracanzani 5 , M. Colombo 1 ,
A. Tripodi 3 , M. Primignani 1
1
U.O. Gastroenterologia-1, IRCCS-Ca’ Granda,
Ospedale Maggiore Policlinico, Università degli studi
di Milano, Milan, Italy
2 U.O. Medicina Interna, IRCCS-San Donato,
Dipartimento di Scienze Biomediche per la Salute,
Università degli studi di Milano, Milan, Italy
3 Centro Emofilia e Trombosi Angelo Bianchi Bonomi,
IRCCS-Ca’ Granda, Ospedale Maggiore Policlinico,
Università degli studi di Milano, Milan, Italy
4 U.O. Gastroenterologia-2, IRCCS-Ca’ Granda,
Ospedale Maggiore Policlinico, Università degli studi
di Milano, Milan, Italy
5 U.O. Medicina Interna, IRCCS-Ca’ Granda, Ospedale
Maggiore Policlinico, Università degli studi di
Milano, Milan, Italy
Introduction: Anticoagulation with vitamin K antagonists
(VKAs) is an effective and relatively safe therapy for patients with
portal vein thrombosis (PVT).
Aim: Exploring the haemorrhagic risk of VKAs in relation with
the presence of cirrhosis.
Material and methods results: Retrospectively, we compared
the VKAs-related bleeding risk in cirrhotic patients with de novo
PVT (PVT-cohort, n = 62) vs non-cirrhotic patients with a thromboembolic event (TE-cohort, n = 160). Any bleeding during 4-years
of follow-up or withdrawal of anticoagulation, was recorded.
Bleeding-risk due to portal hypertension (PHT) in the PVT-cohort
was compared with an independent series of cirrhotics with
PHT unexposed to VKAs during follow-up (CH-cohort, n = 53).
Major bleeding episodes under anticoagulation were intracranial
or retroperitoneal events, fatal bleeding events, need of hospitalization or transfusion, otherwise they were considered minor
bleedings. All patients with cirrhosis were prophylaxed for PHTrelated bleeding according to current guidelines.
TE-cohort and PVT-cohort were comparable for age, sex and
time in therapeutic range (TTR) of the INR. The treatment with VKAs
was longer for the TE-cohort (31.1 ± 16.9 vs 23.0 ± 16.2 months,
p = 0.001). Overall, 41 patients under anticoagulation experienced
a bleeding episode (14 major/27 minor). The actuarial probability
of major/minor bleedings was higher in PVT-cohort (23%/30%) than
in the TE-cohort (6%/20%) (p < 0.001). However, the risk of uppergastro-intestinal bleeding in PVT-cohort (15%) was the same as in
the CH-cohort (13%) also adjusting for potential confounders, confirming that VKAs do not increase the risk of bleeding due to PHT.
Finally, the exclusion of the upper-gastrointestinal bleeding in PVTcohort led to a significant reduction of major bleedings accountable
for VKAs, leaving a significant residual risk only for minor episodes
(p < 0.05).
Conclusions: VKAs expose patients with cirrhosis and PVT to
an additional risk of minor bleedings. This should be taken into
account in future clinical studies to ameliorate the benefit/risk ratio
of anticoagulation in this clinical setting.
H. Stefanescu 1,2 , B. Procopet 3,4 , G. Allegretti 1 ,
A. Berzigotti 3 , N. Gamal 1 , F. Conti 1 , D. Festi 1 ,
P. Andreone 1 , L. Bolondi 1 , J. Bosch 3 , F. Piscaglia 1
1
University of Bologna, Italy
2 Hepatology Department, Regional Institute of
Gastroenterology and Hepatology, ClujNapoca,
Romania
3 Liver Hemodynamic Unit, Hospital Clinic y
Provincial, University of Barcelona, Spain
4 3rd Medical Clinic, University of Medicine and
Pharmacy, ClujNapoca, Romania
Background: Liver and spleen stiffness measurement
(LSM;SSM) are used for the noninvasive assessment of chronic liver
diseases and may identify clinically significant portal hypertension
(CSPH). Besides transient elastography new methods are proposed,
but their performance to assess PH and their reliability criteria are
unknown. We aimed to assess diagnostic accuracy for CSPH and to
determine a set of reliability criteria for bidimensional shear wave
elastography (2D-SWE).
Methods: 51 consecutive cirrhotic patients (61% M; 59.7 yrs)
submitted to HVPG measurement were enrolled. All patients
underwent LSM and SSM by 2D-SWE (Aixplorer, SupersonicImaging). Multiple recordings of LS and SS were performed; the
final result was calculated as their median. Each measurement provides mean stiffness within Region of Interest (ROI) and standard
deviation (SD). The median of individual SD and the interquartile
range (IQR) of measurements for each patient were also calculated.
Results: LSM was successful in 47 patients (92.2%), while SSM
only in 42(82.4%). The median number of valid measurements was
5 (3–11) for LSM and 4 (1–10) for SSM.
Both LSM and SSM correlated well with HVPG (r = 0.663;
p < 0.0001 and r = 0.622; p < 0.0001, respectively). Median LSM and
SSM significantly differed in patients without/with CSPH (15.0 vs.
26.1 kPa; p < 0.0001 and 26.0 vs. 39.3 kPa; p = 0.001, respectively).
LSM and SSM had good performance to predict CSPH:
AUROC = 0.893; accuracy = 75.5% (34/45) and AUROC = 0.837; accuracy = 65% (26/40).
Using IQR < 30% of median LSM as additional reliability criteria,
the accuracy increased up to 85.2% (23/27 patients). No benefit was
observed by increasing the threshold of valid measurements > 3, or
by using stricter SD (<20% or <10% of median LSM).
IQR/median and SD/median SSM (both <0.3) raised the accuracy
to 76.5% (13/17 patients) and 82.3% (14/17 patients), respectively.
Conclusion: LSM and SSM by 2D-SWE have a good applicability in patients with PH and acceptable diagnostic performance for
CSPH. However, the reliability is increased if at least 3 measurements are recorded and if IQR/median < 0.3 is used.
Liver – Annual Meeting 2015: Poster sessions – THURSDAY
T-01
ADRENERGIC HYPERFUNCTION IS A KEY
TRIGGER OF SOLUTE-FREE WATER RETENTION
IN ASCITIC CIRRHOSIS: VASOPRESSIN (ADH) IS
NOT THE ONLY AGENT TO BLAME
G. Sansoè 1 , M. Aragno 2 , R. Mastrocola 2 ,
F. Rosina 1 , M. Parola 2
Conclusions In ascitic cirrhosis, reduced volaemia, adrenergic
hypertone, and secondary aldosteronism, especially when exacerbated by furosemide, contribute to tubular water retention.
Sympatholytic agents are as effective as V2 -antagonists to blunt
tubular water retention.
T-02
1
Division of Gastroenterology, Gradenigo Hospital,
Turin, Turin, Italy
2 Department of Clinical and Biological Sciences,
University of Turin, Turin, Italy
Introduction Adrenergic hyperfunction causes proximal tubular fluid retention and reduces renal excretion of solute-free water,
but, in advanced cirrhosis, non-osmotic hypersecretion of vasopressin (ADH) is considered the cause of water retention and
hyponatremia. Aim. Since ADH V2 receptor antagonists are not
beneficial in long-term treatment of ascites, we hypothesize that
water retention in experimental ascitic cirrhosis depends also on
adrenergic hyperfunction.
Methods Hormonal status, renal function and tubular freewater reabsorption (TFWR) were assessed in 6 groups of rats
with ascitic cirrhosis: rats with cirrhosis due to 13-week CCl4
administration (group G1); cirrhotic rats receiving daily diuretics
(0.5 mg/kg furosemide plus 2 mg/kg K+ -canrenoate) during 11th 13th weeks of CCl4 (G2); cirrhotic rats treated daily with diuretics
associated with guanfacine oral prodrug (␣2A adrenergic receptor agonist and sympatholytic agent) 2 (G3), 7 (G4), or 10 mg/kg
(G5), or SSP-004240F1 (V2 receptor antagonist) 1 mg/kg (G6) during
11th -13th weeks of CCl4 .
Results Sodium excretion was lower in G1 than G2-G4 and
G6 (all P < 0.05). TFWR was higher in G1 (untreated cirrhosis, 32 ± 11 microL/min) than in G6 (diuretics + V2 -antagonists,
21 ± 9 microL/min) or G3 (diuretics + guanfacine 2 mg/kg, 20 ± 8
microL/min) (all P < 0.03). Guanfacine, added in G3 to diuretics (G2),
reduced serum norepinephrine from 423 ± 122 to 211 ± 111 ng/L
(P < 0.01), plasma renin activity from 25 ± 12 to 9 ± 7 ng/mL/h
(P < 0.03), and TFWR from 45 ± 18 to 20 ± 8 microL/min (P < 0.01).
Among all rats, TFWR did not correlate with ADH levels (r = 0.02, P:
n.s.), but did with plasma aldosterone (r = 0.51, P < 0.01) and urinary
potassium excretion (r = 0.90, P < 0.001).
1590-8658/$36.00
CHRONIC PARTICULATE MATTER EXPOSITION
AND WESTERN DIET INDUCE THE PROGRESSION
FROM STEATOSIS TO STEATOHEPATITIS
M. Tarocchi 1,∗ , G. Marroncini 1 , S. Polvani 1 ,
S. Tempesti 1 , E. Ceni 1 , T. Mello 1 , M. Peluso 2 ,
A. Galli 1
1 Department of Experimental and Clinical
Biomedical Sciences, University of Florence, Florence,
Italy
2 Cancer Risk Factor Branch, Cancer Prevention and
Research Institute, Florence, Italy
Introduction: Nonalcoholic fatty liver disease (NAFLD) is the
most common chronic liver disease, and the prevalence is rapidly
increasing in developed countries. Nonalcoholic steatohepatitis
(NASH), the severe form of NAFLD, can also progress to liver cirrhosis and hepatocellular carcinoma. Recent evidences suggest that
environmental factors can trigger hepatic inflammation and progression of steatosis to NASH.
Aim: We evaluate if a western style diet in association with
chronic urban particulate matter exposition can modifies the
pathogenesis and progression of NASH.
Materials and methods: The experimental model was created to
reproduce urban lifestyle: C57Bl/6 mice were fed with a western
style diet (HFD), and treated with particular matter (PM) collected
from the urban area of Florence (Italy). After 4 and 8 weeks were
performed the morphologic analysis of liver tissues, the evaluation
of inflammatory cell infiltrate and the collagen deposition; we evaluate also the effects of PM on cytokine production and oxidative
stress related cellular damage.
Results: Both the HFD groups developed fat accumulation in the
liver, and at 8 weeks, the NASH score was significantly increased
in HFD-PM group (p < 0.01) based on the histological analysis,
the tissue fat content, the inflammatory cell infiltrate and the
e20
collagen deposition. The cytokine profiles indicate an increased
production of pro-inflammatory molecules in presence of PM. The
levels of DNA adducts were significantly increased in HFD groups
and,interestingly, the effects of HFD and PM were synergistic in fact
the DNA adducts were about thirty fold greater in HFD mice compared to controls treated with the same amounts of air particulate
matters.
Conclusions: Our data suggest that in subjects with steatosis
due to a western style of life, the association with a chronic exposition to urban particulate matter plays an important role in the
pathogenesis of NASH and its evolution to cirrhosis and also to
cancer.
early systolic dysfunction. Similarly, ESV was increased in patients
with fibrosis (69.2 ± 16.9 vs 94.5 ± 31.4cc, p = 0.018), whereas
ejection fraction (51 ± 7 vs 59 ± 7%, p = 0.034) and cardiac index
(2.9 ± 0.7 vs 3.8 ± 0.9, p = 0.03) were significantly reduced.
Conclusions: In NAFLD subjects metabolic derangements and
histological features are associated with early systolic LV dysfunction, independently of diabetes, hypertension and dyslipidemia.
Funded by FP7/2007-2013 under grant agreement n HEALTHF2-2009-241762 for the project FLIP and by PRIN 2009ARYX4 T
NLRP3 INFLAMMASOME INCREASES HEPATIC
FIBROSIS BY INDUCING INFLAMMATORY
SIGNALS IN HEPATIC STELLATE CELLS
T-03
INSULIN RESISTANCE AND LIVER DAMAGE ARE
ASSOCIATED WITH EARLY SIGNS OF LEFT
VENTRICULAR SYSTOLIC DYSFUNCTION IN
NON-DIABETIC, NON-DYSLIPIDEMIC,
NORMOTENSIVE PATIENTS WITH
NONALCOHOLIC FATTY LIVER DISEASE
E. Vanni 1,∗ , L. Mezzabotta 1 , R. Faletti 2 ,
M. Morello 3 , A. Marengo 1 , G. Battisti 2 , S. Frea 3 ,
M. Cannillo 3 , C. Rosso 1 , E. Mosso 1 ,
L. Bergamasco 4 , M. Rizzetto 1 , E. Bugianesi 1
1 Department of Medical Sciences, Division of
Gastroenterology, University of Turin, Turin, Italy
2 Department of Diagnostic Imaging and
Radiotherapy, Radiology, University of Turin, Turin,
Italy
Cardiology, University of Turin, Turin, Italy
4 University of Turin, Turin, Italy
Background andAims: Nonalcoholic Fatty Liver Disease (NAFLD)
has been associated with subclinical cardiovascular disease (CVD).
This study was undertaken to evaluate the relationship between
metabolic parameters, histologic features and parameters of cardiac morphology and function in NAFLD subjects.
Methods: Nineteen non-diabetic, non-dyslipidemic, nonhypertensive patients with biopsy-proven NAFLD (17 men, 41 ± 8
years, BMI 26.8 ± 3 kg/m2 ) and 9 healthy controls (5 men, 30 ± 2
years, BMI 22.5 ± 2 kg/m2 ) underwent transthoracic echocardiography and cardiac-MRI to evaluate cardiac morphology and
function. Endogenous glucose production (EGP) and lipolysis were
assessed by stable isotope tracers. Hepatic Insulin resistance (IR)
as EGPxfasting insulin, Oral Glucose Insulin Sensitivity (OGIS),
adipo-IR as free fatty acids (FFAs)xfasting insulin were calculated.
Results: NAFLD patients had significantly higher concentration of FFAs than controls (p < 0.05) and higher total
saturated and monounsaturated levels (p < 0.05). In NAFLD
basal Hepatic-IR (NAFLD vs controls: 92 ± 34 vs 52 ± 18
umol/minkg*mU/L) and Adipo-IR (NAFLD vs controls: 21 ± 10
vs 11 ± 5 mmol/L*mU/L) were significantly increased (p < 0.03 for
all) and OGIS significantly reduced (NAFLD vs controls: 11.0 ± 1.64
vs 13.1 ± 1.1 mg/kgmin, p = 0.005). The end-systolic LV diameter
(30.4 ± 3.7 vs 27.2 ± 3.5 mm, p = 0.044) was significantly higher
in patients than in controls. In NAFLD patients, both hepatic-IR
and adipo-IR directly correlated with MRI end systolic LV volume
(ESV) (r = 0.63, p = 0.004 and r = 0.54, p = 0.018, respectively), while
OGIS was inversely related to end-systolic LV diameter (r = -0.48,
p = 0.037) and ESV (r = -0.48, p = 0.036). At liver biopsy, steatosis
≥33% was associated with increased ESV (p = 0.047), suggesting
T-04
C. Rychlicki ∗ , L. Agostinelli, E. Mingarelli,
S. Saccomanno, C. Pinto, I. Pierantonelli, L. Trozzi,
A. Benedetti, M. Marzioni, S. De Minicis,
G. Svegliati-Baroni
Department of Gastroenterology, Polytechnic
University of Marche, Ancona, Italy
Background and Aims: Hepatic fibrosis represents the woundhealing response process to chronic liver injury, independently
from aetiology. Hepatic stellate cells (HSCs) are the main liver extracellular matrix producing cells and also exert proinflammatory
activity. The NLRP3 inflammasome mediates the release of proinflammatory cytokines in response to cellular danger signals in
various organs, but its role in the progression of hepatic injury is
still unclear. Thus, aim of the study was to evaluate the role of NLRP3
inflammasome in fibrosis development and in HSCs behaviour.
Methods: Wild-type (WT) C57BL/6 and Nlrp3A350VneoR (Nlrp3-/− )
mice underwent bile duct ligation for 3 weeks. Quiescent HSCs
(quHSCs) and in vitro activated HSCs (AcHSCs) from WT mice were
obtained respectively after 12 hours and 6 days of culture.
Results: Compared to WT, Nlrp3-/- BDL mice showed a significant
reduction in liver inflammasome activation, associated to significantly decreased TLR4, TLR9 and IL6 mRNA levels. Type I collagen,
TGF␤, CTGF and TIMP1 gene expression were significantly reduced
in Nlrp3-/- BDL mice, leading to reduced collagen deposition measured by Sirius Red staining. In vitro HSCs activation was associated
to significantly decreased gene expression of inflammasome components compared to (qu)HSCs. However, (Ac)HSCs stimulation
with LPS induced expression of the inflammasome pathway, without affecting neither type I collagen mRNA nor cell proliferation.
After priming with LPS, incubation with ATP was needed for the
release of IL1␤. (Ac)HSCs incubation with recombinant IL1␤ further stimulated the inflammasome pathway and led to increased
TGF␤ and CTGF gene expression and HSCs migration.
Conclusions: NLRP3 inflammasome contributes to hepatic fibrosis by increasing the inflammatory response and inducing a
paracrine loop that maintains TLRs activation and IL6 production.
IL1␤ released by (Ac)HSCs stimulates fibrogenesis and cell migration in an autocrine manner. Several new therapeutic targets can
be identified in the NLRP3 inflammasome pathway to treat hepatic
fibrosis.
T-05
A NOVEL ROLE FOR THE KYNURENINE
PATHWAY IN EXPERIMENTAL STEATOHEPATITIS
E. Vivoli 1 , A. Cappon 1 , A. Cozzi 1 , N. Navari 1 ,
M. Gargano 2 , F. Fallarino 2 , F. Marra 1
1
Dipartimento di Medicina Sperimentale e Clinica,
2 Medicina Sperimentale, University of Perugia,
Perugia, Italy
Background and Aims: The pathogenic mechanisms underlying development of nonalcoholic steatohepatitis are still elusive.
Indoleamine 2,3-dioxygenase (IDO-1), an enzyme that mediates
the catabolism of L-tryptophan to L-kynurenine, plays an important role in hepatic immune regulation, mediating inflammation or
tolerance depending on the type of injury and the tissue involved.
In the present study, we examined the effect of pharmacological or
genetic inhibition of IDO-1 on the development of steatohepatitis
induced by a methionine and choline-deficient (MCD) diet in mice.
Methods: Balb/C mice fed a MCD diet for 8 weeks, were
treated with the specific IDO inhibitor, 1-methyl-D-triptophan
(1MT, 5 mg/ml) or its vehicle in drinking water. WT and IDO-1 KO
C57Bl/6 mice were fed MCD or its control for 4 weeks. Intrahepatic
gene expression was assayed by quantitative real time PCR.
Results: 1MT administration caused a significant reduction of
ALT levels in MCD-fed mice, while no changes were observed
comparing the WT + MCD and IDO-1 KO + MCD groups. Both pharmacologic and genetic interference with IDO-1 resulted in an
amelioration of the inflammatory phenotype. These effects were
associated with reduced intrahepatic expression of proinflammatory factors, including CD11b, CCL2, TNF␣, and IL-1␤. Fibrosis
induced by the MCD diet was decreased by 1MT co-administration
(performed for 8 weeks), together with reduced intrahepatic gene
expression of TGF-␤ and ␣-SMA.
Conclusions: Interference with the IDO pathway by 1-MT
administration or by genetic deletion of IDO-1 ameliorates the phenotype of dietary experimental steatohepatitis and modulates the
expression of proinflammatory and profibrogenic factors in this
context.
T-06
REDUCTION IN SUMOYLATION-DEPENDENT
S100A4 NUCLEAR IMPORT IN
CHOLANGIOCARCINOMA BY LOW DOSE
PACLITAXEL HALTS TUMOR INVASIVENESS AND
HEMATOGENOOUS METASTASIZATION BY
DOWN-MODULATING RHO-A AND CDC42
ACTIVITIES
G. Spagnuolo 1,∗ , M. Cadamuro 1,2 , L. Sambado 1 ,
S. Indraccolo 3 , G. Nardo 3 , A. Rosato 3 , E. Novelli 4 ,
C. Spirli 5 , M. Strazzabosco 1,5 , L. Fabris 1,2,5
1 Dep. of Surgery & Translational Medicine,
University of Milan-Bicocca, Italy
2 Dep. of Molecular Medicine, University of Padua,
Italy
3 Istituto Oncologico Veneto, IRCCS, Padua, Italy
4 Clinica San Gaudenzio, Novara, Italy
5 Section of Digestive Diseases, Yale University, USA
Background. Cholangiocarcinoma (CCA) is characterized by
early and strong invasiveness. Nuclear expression of the S100A4
e21
protein is a marker of increased CCA invasiveness and our preliminary data showed that Paclitaxel (PTX) could reduce S100A4
nuclearization. We aimed at studying if nuclear S100A4 promotes
CAA invasiveness and may represent a therapeutic target.
Methods and Results. CCA cells expressing nuclear S100A4
(EGI-1) were treated with increasing PTX doses to study its effects
on nuclear S100A4 expression, S100A4 sumoylation (a mechanism
of nuclear import of proteins), cytoskeletal integrity, cell proliferation/apoptosis, motility, invasiveness and Rho GTPases activity.
PTX (1.5 and 15 nM) induced a marked reduction in nuclear S100A4.
This decrement was linked with a significantly reduced sumoylated
fraction and an attenuation of cell migration, invasiveness and
Rho-A/Cdc42 activation, without affecting proliferation/apoptosis
or cytoskeletal integrity. SCID mice xenografted with EGI-1 cells
by spleen injection, were treated, after tumor engraftment, with
low-dose metronomic regimen of PTX (2,6 mg/kg/die; n = 5) for
14 days, using untreated mice as controls (n = 5). After mice
sacrifice, we evaluated PTX effects on tumor size, number of
micrometastasis (MM) and isolated tumour cells (ITC) in liver
and lung samples, along with proliferation, apoptosis and S100A4
expression of EGI-1. PTX induced a significant reduction in both the
tumor size and number of lung MM/ITC. The primary CCA mass of
PTX-treated mice, showed a significantly reduced number of EGI-1
cells expressing nuclear S100A4, whereas proliferation/apoptosis
rate did not change.
Conclusion. Down-regulation of nuclear S100A4 by PTX at doses
below those commonly used in chemotherapy but able to interfere
with the sumoylation process, results in a decreased CCA cell motility and invasiveness and in a reduction of hematogenous spread.
These effects are not dependent upon changes in cell proliferation,
apoptosis or cytoskeleton integrity, but upon down-modulation of
Rho-A and Cdc42 activities by a reduced S100A4 nuclearization.
T-07
COMPLICATIONS AFTER PERCUTANEOUS
RADIOFREQUENCY ABLATION (RFA) OF
HEPATOCELLULAR CARCINOMA (HCC) IN
CIRRHOSIS: 20 YEARS EXPERIENCE IN A SINGLE
CENTER
A. Giorgio 1,∗ , G. Iaquinto 2 , L. Montesarchio 1 ,
P. Gatti 3 , B. Santoro 4 , F. Amendola 5 ,
P. Matteucci 6 , C. Coppola 2,7 , V. Giorgio 7,8
1 Interventional Ultrasound Unit, Tortorella Clinical
Institute, Consorzio ISMESS, Salerno, Italy
2 Unità Operativa di Medicina Interna, Casa di Cura
S. Rita, Atripalda, Avellino
3 Unità Operativa di Medicina Interna, Ospedale di
Fasano, Brindisi, Italia
4 Unità operativa ecografia interventistica, Athena,
Caserta, Italia
5 Unità Operativa di Medicina Interna, Tortorella
Clinical Institute, Consorzio ISMESS, Salerno, Italy
6 Campus biomedico, Roma, Italia
7 Unità Operativa di Medicina Interna, Epatologia
interventistica, ospedale di Gragnano, Napoli, Italia
8 Università Cattolica del Sacro Cuore, Policlinico
Gemelli, Roma, Italia
Aim: To report 20 years experience on complications after RFA
of HCC in cirrhotics, giving special emphasis on the changes of Child
Class, haemorragic events and deaths.
Material and Methods: From April 1994 to March 2014, 1787
RFA procedures were performed percutaneously under ultrasound
e22
guidance in 1162 consecutive cirrhotics (57-85 years; mean 68;
882 males; 852 Child A, 310 Child B). Diameter of HCC nodules ranged from 1.2 to 6.8 cm. ProtrombinTime (PT) and Platelets
Count were > 50% and 50.000 mm3, respectively; total bilirubin
ranged from 0,80 and 3.4 mg/dl (mean 1.6). Three RFA devices
were employed: hooks elecrtrode-needle (170 cases), perfused
electrode-needle (1041 cases) and cold electrode- needle (476
cases). 67 cirrhotics underwent RFA on both intraparenchymal HCC
nodule and tumor thrombus in the main portal vein (PVTT).
Results: Mortality. 4 patients (0,4%) died after RFA: 2 for
haemoperineum, 1 for haemotorax, 1 for liver failure. No patient
treated for both intraparenchimal HCC and PVTT died.
Morbidity. Changes in Child Class of cirrhosis were observed in
19 patients (1.6%): 8 Child A cirrhotics had development of liver
decompesation (ascites and/or jaundice and/decreased PT) changing from Child A to Child B class in 7, and from Child A to Child
C in 1 case, respectively. 11 Child B patients changed to Child C
Class. 6 patients had haemoperitneum with no death. Abscess formation was observed in 2 patients. One patient had an intraepatic
haematoma, that resolved spontaneously.
Conclusion: RFA of HCC in cirrhotic patients can be considered safe, even in case of advanced disease, such as invasion
of portal venous system. Care must be taken in the evaluation
and surveillance of functional liver reserve. Haemorragic adverse
events remain the leading cause of mortality.
T-08
PHARMACOLOGICAL INHIBITION OF SIRT1
PROMOTES APOPTOSIS AND SENESCENCE OF
HEPATIC STELLATE CELLS DURING LIVER INJURY
M. Tarocchi ∗ , G. Marroncini, E. Ceni, S. Polvani,
S. Tempesti, T. Mello, A. Galli
Department of Experimental and Clinical Biomedical
Sciences, University of Florence, Florence, Italy
Introduction: Hepatic fibrosis is characterized by excessive
deposition of extracellular matrix (ECM) which leads to alterations
in liver function. Activated hepatic stellate cells (HSC) are considerate to be the key cell type responsible for excessive ECM deposition
after liver injury. Modulation of SIRT1, a NAD-dependent histone
deacetylase, seems to regulate cellular metabolism, senescence and
apoptosis.
Aim: We investigated the role of SIRT1 activation in HSC during
liver injury as a possible therapeutic target for liver fibrosis.
Materials and methods: We evaluated the expression of SIRT1
in human tissues; we evaluated its role in response to stress conditions in primary human and mouse HSC. We also tested the in vivo
effects of SIRT1 inhibition in different mouse models of liver injury
induced by carbon tetrachloride administration and bile duct ligation.
Results: We found that SIRT1 is constitutively expressed in HSC,
and in higher levels compare to other hepatic cells. In vitro, we
demonstrated that starving, oxidative stress or hypoxia resulted
in SIRT1 increased activity and protected HSC in stress conditions.
Conversely its pharmacological inhibition reduced HSC lifespan due
to higher susceptibility to pro-apoptotic stimuli and the acquisition
of a senescent phenotype. Selective SIRT1 inhibition in mouse models of liver injury lowered the activated HSC specific genes as aSMA,
MMPs, TIMPs and pro-inflammatory cytokines. This was mainly
due to a decrease in activated HSC content in the parenchyma. Interestingly this alteration in the hepatic cellular composition resulted
in a reduction in collagen deposition.
Conclusions: Selective SIRT1 inhibition in vitro increases stressrelated senescence and apoptosis in primary HSC; the modulation
of SIRT1 in the animal model liver decreases the number of activated and proliferating HSC with a consequent reduction of ECM
deposition. The inhibition of SIRT1 could therefore prevent the
fibrogenic alteration of the parenchyma during liver injury and
promote the recovery process.
T-09
JNK SIGNALING ACTIVATED BY
PLATELET-DERIVED GROWTH FACTOR D
(PDGF-D) STIMULATES SECRETION OF VASCULAR
ENDOTHELIAL GROWTH FACTOR-C (VEGF-C) BY
CANCER-ASSOCIATED FIBROBLASTS TO
PROMOTE LYMPHANGIOGENESIS AND EARLY
METASTATIZATION IN CHOLANGIOCARCINOMA
M. Cadamuro 1,2 , M. Vismara 1 , S. Brivio 1 ,
A. Furlanetto 3 , M. Strazzabosco 1,4 , L. Fabris 1,2,4
University of Milan-Bicocca, Milan, Italy
Padua, Padua, Italy
3 Pathology Unit, Treviso Regional Hospital, Italy
4 Section of Digestive Diseases, Yale University
School of Medicine, New Haven, Connecticut, USA
Background: Cholangiocarcinoma (CCA) is a highly invasive
malignancy with a poor prognosis. Currently, < 30% of CCA patients
undergo surgical resection, due to the early lymph node metastasization. In CCA, lymphangiogenesis develops within an abundant
stroma, mainly composed by cancer-associated fibroblasts (CAF)
recruited by Platelet-derived Growth Factor-D (PDGF-D) secreted
by CCA cells. Mechanisms governing lymphangiogenesis in CCA are
unknown. We aimed at investigating the role of PDGF-D-mediated
epithelial-mesenchymal interactions in promoting lymphangiogenesis.
Methods/Results: Immunohistochemistry on human CCA specimens (n = 6), were performed to understand the relationship
among lymphatic vessels (D2-40+ ), blood vessels (CD34+ ), CCA and
CAF (␣SMA+ ), and the expression of PDGF-D, PDGFR␤, VEGF-C and
VEGFR3. Furthermore, lymphatic (LMVD) and vascular microvessel density (VMVD) were calculated in CCA and compared with
hepatoma (n = 6). With respect to hepatoma, CCA was characterized by an increased LMVD and a reduced VMVD. In CCA,
lymphatic endothelial cells (LEC) laid in close vicinity to CAF and
to cancer cells. We observed a specific expression of PDGF-D (on
CCA cells), PDGFR␤ and VEGF-C (CAF), and VEGFR-3 (LEC), suggesting a sequential CCA cells-CAF-LEC crosstalk. To verify this,
we measured in primary fibroblasts challenged with PDGF-D the
secretion of lymphangiogenic growth factors (VEGF-C, Ang-1 and
Ang-2) (ELISA). PDGF-D stimulated fibroblasts to secrete VEGF-C
but not Ang-1 (Ang-2 was not expressed), an effect significantly
reduced by PDGFR␤ (imatinib mesylate), and JNK (SP600126)
inhibitors. Using Boyden chambers, we evaluated the motility
of human LEC following stimulation with conditioned medium
of PDGF-D-challenged fibroblasts with/without SP600126. Conditioned medium induced a significantly stronger effect on LEC
recruitment than that of unstimulated fibroblasts, an effect significantly hampered by SP600126.
Conclusions: PDGF-D released by CCA cells stimulates CAF to
secrete VEGF-C through JNK signaling. In turn, VEGF-C secreted by
CAF stimulates LEC recruitment. This cross-talk identifies PDGFR␤
and JNK as targets for antilymphagiogenic therapies in CCA.
T-10
THE AMPK RELATED KINASE NUAK2 INTERACTS
WITH TGF-BETA AND REGULATES THE
ACTIVATION PROCESS OF HEPATIC STELLATE
CELLS (HSC)
A. Provenzano 1,∗ , C. Tosti Guerra 1 , A. Caligiuri 1 ,
K. Rombouts 2 , M. Pinzani 2 , F. Marra 1
1 Dipartimento di Medicina Sperimental e Clinica,
2 Institute for Liver and Digestive Health, University
College London, London, United Kingdom
Background and Aims: Nuak2 is a member of AMPK related
kinases (ARKs), that act as energy sensors and controllers of cellular
structure, with different effects on cell motility and cytoskeletal
organization, depending on the cell types. AMPK possesses antifibrogenic activity. A recent study showed a link between Nuak2
and TGF␤, a major driver of hepatic fibrogenesis-. However, little
information is available on the role of Nuak2 in liver fibrosis, and
in particular on HSC trans-differentiation. Aims of this study was
to elucidate the involvement of Nuak2 in HSC transactivation, and
to investigate the possible interaction between Nuak2 and TGF␤
signaling.
Methods: HSC were isolated from normal rat and human liver
and activated by culture on plastic. Knockdown of Nuak2 was
achieved by siRNA. Cell migration was evaluated in modified Boyden Chambers. Protein expression and signaling pathways were
analyzed by Western blotting.
Results: In fully activated HSC, down-regulation of Nuak2
positively modulated cell migration and induced changes in the
expression of molecules involved in cytoskeletal organization.
Knockdown of Nuak2 increased ␣-SMA and p-SMAD3 expression,
in HSCV exposed to TGF␤. Moreover, Nuak2 was up-regulated in
HSC following TGF␤ treatment.
Conclusions: The AMPK related kinase, Nuak2, is modulated
during the activation process of HSC, regulates cytoskeletal organization and cell motility and interacts with TGF-␤ in regulating the
pro-fibrogenic properties of HSC.
e23
T-11
THE ITA.LI.CA STAGING SYSTEM FOR PATIENTS
WITH HEPATOCELLULAR CARCINOMA: A
MULTICENTER COHORT STUDY
F. Farinati 1,1 , A. Vitale 1,1 , F. Trevisani 2 , T. Huo 3,4 ,
Y.-H. Lee 3,4 , U. Cillo 1 , On behalf of the ITA.LI.CA.
study group
1 Department of Surgery, Oncology and
Gastroenterology, University of Padua, Padua, Italy
2 Department of Medical and Surgical Sciences,
Division of Semeiotics, Alma Mater
Studiorum–University of Bologna, Bologna, Italy
3 National Yang-Ming University, Taiwan
4 Department of Medicine, Taipei Veterans General
Hospital, Taipei, Taiwan
Background The Barcelona Clinic Liver Cancer (BCLC) classification ability to stratify HCC patient survival still remains
controversial when compared to that of other systems (HKLC,
MESIAH, CLIP, JIS). The aim of this study is to develop and validate
a new staging system for HCC.
Methods The ITA.LI.CA (Italian Liver Cancer) database was
divided in training (n = 3628, 70%) and validation (n = 1555, 30%)
cohorts. A further cohort of 2651 HCC patients from Taiwan was
used for external validation. We defined the ITA.LI.CA Tumor Status (TS) using BCLC stages (0, A, B, C) but adding 3 intermediate
B sub-stages: B1 = single > 5 cm or 2-3 nodules 3-5 cm; B2 = 23 nodules > 5 cm or > 3 nodules ≤ 5 cm; B3 = > 3 nodules > 5 cm or
presence of intrahepatic macrovascular invasion. TS was then
included in a multivariate survival model together with Child
Pugh Score (CPS) and ECOG performance status (PST), and a novel
ITA.LI.CA prognostic staging was developed.
Results The ITA.LI.CA staging system was based on the following stages: 0n, An, B1n, B2n, B3n, Cn, where capital letters defined
TS stages indicating potential for treatment as in BCLC (from radical to palliative therapies), while n defined an additional score
(0/1 = eligible for HCC therapy; 2/ > 2 = contraindication to therapy)
based on CPS and PST indicating treatment feasibility. The concordance (c)-statistics for this model in training, internal and external
validation cohorts were respectively 0.72, 0.71, and 0.78, and they
were superior to that for BCLC (0.65, 0.64, and 0.73), CLIP (0.69,
0.68, and 0.75), JIS (0.67, 0.67, and 0.70), MESIAH (0.69, 0.69, and
0.77), and HKLC (0.68, 0,68, and 0.75).
Conclusion We developed a novel ITA.LI.CA staging system, easy
to apply in clinical practice, and with a strong prognostic impact in
two large western and eastern populations.
1
These authors equally contributed to this work.
e24
T-12
T-13
MIR-106B FOR HCC PATIENT MANAGEMENT
AFTER THERAPY
OLEUROPEIN REDUCES INFLAMMATORY
MEDIATORS AND HEPATIC IMMUNE CELLS
INFILTRATION IN A MOUSE MODEL OF NAFLD
D. Pascut 1,∗ , R. Patti 2 , N. Mezzina 3 , C. Abazia 3 ,
F. Masutti 3 , S.L. Crocè 2,3 , C. Tiribelli 1,2,3
1
Fondazione Italiana Fegato, AREA Science Park
Basovizza, Trieste, Italy
2 Department of Medicine, Surgery, and Health
Sciences, University of Trieste, Trieste, Italy
3 Department of General Surgery, Teaching Hospital
Cattinara, Trieste, Italy
Background: MicroRNAs (miRNAs) are regulatory noncoding
RNAs. Their aberrant expression is observed in many diseases,
including Hepatocellular carcinoma (HCC). They are stable in biological fluids making miRNAs promising class of potential non
invasive blood biomarkers for patients’ follow-up. mir-106b is frequently de-regulated in HCC with a role in cell proliferation and
migration. Recent findings reported a circulating mir-106b altered
expression in HCC.
Aim: To investigate the role of miR-106b as peripheral blood
biomarker in HCC.
Methods: 15 consecutive patients with early/intermediated
stage HCC were enrolled. Patients were treated according to the
ESSL/AASLD practice guidelines. All patients were staged at time 0,
1 and 6 months after therapy with CT scan and/or MRI. The longest
follow-up was 20 month. Stabilized Pax-gene tubes where used
to collect total blood before (T0) and one month (T1) after the
treatment. Small RNA were purified and quantitatively analyzed
by RT-qPCR. The relationships between their expression levels
and clinicopathological parameters were further determined. The
Kaplan-Meier model was used to estimate disease-free survival
(DFS).
Results: A correlation between miRNA levels and treatment
response was observed (P < 0.001), 60% of the patients showed a
complete responseto treatment with a median mir-106b expression of 2.4 at T1. The 40% showed a partial response or progressive
disease (median mir-106b expression = 0.78). Kaplan-Meier estimates and the log-rank test showed that high expression of
mir-106b correlated with the longest DFS (P < 0.0038). Patients
(n = 8) with higher miR-106b expression had a longer DFS time
(median = 15.5 months) as compared to patients (n = 7) with lower
expression (median = 1 months). The expression of miRNA-106b
was significantly correlated with the with BCLC staging A1, A2 and
A3 (P < 0.001).
Conclusions: Circulating mir-106b detection appears as a
promising non invasive marker to identify patients with longer DFS
in response to anticancer therapies.
M. Arciello 1,2,∗ , B. Barbaro 1 , A. Longo 1 ,
R. Maggio 1 , C. Viscomi 1 , C. Balsano 1,3
1 Laboratory of Molecular Virology and Oncology,
2 Department Internal Medicine and Medical
Specialties, “Sapienza” University of Rome, Rome,
Italy
3 Institute of Molecular Biology and Pathology
(IBPM) - CNR, Rome, Italy
Introduction: The metabolic syndrome (MeS) is a cluster of
metabolic abnormalities like obesity, insulin-resistance and cardiovascular disease, with a growing worldwide prevalence that
reaches about 40% in population over 50 years of age. Nonalcoholic fatty liver disease (NAFLD) is considered a pathogenic factor
of MeS as well as its hepatic manifestation. In some cases NAFLD
may progress from simple steatosis to end-stage liver diseases.
Olives, main component of the Mediterranean diet, exerts beneficial effects on liver and heart, and reduces inflammation in MeS
patients, probably due its polyphenols such as oleuropein.
Aim: We aim to assess whether oleuropein supplementation
may counteract metabolism alterations and inflammation produced by an excessive fat intake.
Methods: As model for NAFLD we used C57BL/6 mice fed with
a high fat diet (HFD). After 8 weeks of HFD feeding, mice received
a HFD supplemented with 3% oleuropein (OLE) for further 8 weeks
(HFD + OLE).
Results: After preliminary evaluation of oleuropein efficacy in
in vitro model of steatosis, we evaluated its effects in mouse model
of NAFLD. After 16 weeks, HFD-fed mice show elevated fat deposition, increased body (BW), liver (LW) and heart (HW) weights and
an increase of several circulating cytokines. HFD + OLE mice show
reduced weight gain (BW - 25%, LW–50%, HW–70%) compared to
HFD-fed mice, reduced liver damage and inflammatory infiltration. Moreover, using the Bio-Plex multiplex biometric ELISA-based
immunoassay in HFD + OLE we appreciated a significant reduction
of various cytokines, including monocyte chemoattractant protein
1 (MCP1) and chemokine (C-X-C motif) ligand 1 (CXCL1). Interestingly, MCP1 and CXCL1 are renowned players in the recruitment of
immune cells and their increase is correlated to MeS.
Conclusion: These results suggest that oleuropein may prevent
NAFLD progression and MeS occurrence counteracting immune
cells infiltration in the liver, a key event in the progression of hepatic
damage.
T-14
T-15
INCIDENCE AND RISK FACTORS OF
HEPATOCELLULAR CARCINOMA IN UNTREATED
SUBJECTS WITH CHRONIC HEPATITIS B VIRUS
INFECTION: A SYSTEMATIC REVIEW AND
META-ANALYSIS
ADHERENCE TO AISF RECOMMENDATIONS FOR
INTEGRATED MANAGEMENT OF
HEPATOCELLULAR CARCINOMA IN A
REAL–WORLD CLINICAL PRACTICE
E. Raffetti 1,∗ , G. Fattovich 2,3 , F. Donato 1
1 Unit of Hygiene, Epidemiology and Public Health,
University of Brescia, Brescia, Italy
2 Department of Medicine, University of Verona,
Verona, Italy
3 Clinical Unit of Gastroenterology, Azienda
Ospedaliera Universitaria Integrata Verona, Verona,
Italy
Introduction and aims. The hepatocellular carcinoma (HCC) risk
in chronic hepatitis B virus (HBV) infection may vary in different
clinical settings. We aimed to assess incidence rates, and factors
related with occurrence, of HCC in untreated subjects with HBV
chronic infection.
Materials and Methods. We performed a systematic review and
meta-analysis searching for published articles in Medline, Embase,
and the Cochrane Library up to October 2014 as well as in reference literature. We included longitudinal studies assessing HCC
incidence in untreated HBV infected patients. The HCC incidence
rates (IRs) estimates and their 95% confidence intervals (95% CIs)
were extracted by each study and pooled together in random effects
models. Assessment of heterogeneity and meta-regression analyses were done.
Results. 79 studies were included with a total of 363,838 participants and 3,472 HCC cases. In asymptomatic carries, the HCC
IRs were 0.04 (95%CI: 0.012-0.07), 0.19 (0.07-0.31) and 0.44 (0.230.66) per 100 person-years in Europe, North America (around 70%
Asiatic subjects) and East Asia, respectively. In inactive carriers and
subjects with chronic hepatitis, the HCC IRs were 0.02 (0.0-0.06)
and 0.12 (0.03-0.27) in Europe, and 0.05 (0.03-0.06) and 0.49 (0.320.66) in East Asia, respectively. In Child-Pugh A cirrhosis, IRs were
2.03 (1.30-2.77), 2.89 (1.23-4.55) and 3.62 (2.65-4.58) in Europe,
North America and East Asia, respectively. Meta-regression analysis showed a statistically significant IR increase with increasing age
(every 10 years: 1.09; 1.05-1.14), male percentage (1.02; 1.00-1.05),
in North America (3.10; 1.00-9.98) and East Asia (2.24; 1.04-4.82)
with respect to Europe, and in asymptomatic carriers (1.84; 0.923.66), and patients with chronic hepatitis (7.84; 3.44-17.98) and
compensated cirrhosis (18.92; 9.93-35.71) with respect to inactive
carriers.
Conclusions. The risk of developing HCC in untreated subjects
with HBV chronic infection is strongly influenced by age, gender,
macro-area of origin, and HBV liver disease status.
e25
G. Piai 1 , V. Messina 2 , G. Valente 1 , L. Rinaldi 1 ,
G. Moggio 3
1 UOSD Fisiologia Epatica con Servizio di Assistenza
ai Trapiantandi e Trapiantati Epatici, AORN
Sant’Anna e San Sebastiano, Caserta, Italy
2 UOS Ottimizzazione e Monitoraggio delle Epatiti
Croniche Virali UOC Malattie Infettive, AORN
Sant’Anna e San Sebastiano, Caserta, Italy
3 UOSD Radiologia Interventistica, AORN Sant’Anna e
San Sebastiano, Caserta, Italy
Introduction and Aim: AISF recommendations (AISF-R) for
HCC management are based on the assumption of a multidisciplinary approach in a network with availability of all diagnostic
and therapeutic resources. In our real world, however, a specialized
hepatic surgery unit is missing. Hepatologists, radiologists, oncologists were then all invited to join a multidisciplinary HCC-team that
we connected to many centres of liver surgery throughout Italy.
Aim of study was to evaluate the efficiency of this organized
application of AISF-R, considered as a methodological effort to
improve quality of assistance and to reduce the high health mobility
from our geographic area.
Methods: One hundred twenty-six HCC patients, stratified
according to BCLC system, were discussed by HCC-team (January
2012 - September 2014). Each formal clinical record was then independently revised, expressing a “theoretical” indication according
to AISF-R and comparing it to the really applied choice.
Results: In 77 cases surgery appeared obviously not indicated
or contraindicated, while liver surgery advice was requested in 49
cases (39%): subdivided in BCLC A1-A3/A4/B/C/D groups, rates were
100%, 2%, 37%, 26%, 17%, respectively.
Overall, 69% of patients were treated according to AISF-R. Adherence was 11/12 (92%) early HCC theoretically eligible for resection,
10/21(48%) liver transplantation, and 21/28 (75%) percutaneous
treatments (PT). TACE was performed in 12/61 (20%) early HCC,
in 3/31 (10%) advanced HCC, and excluded in 14/34 (41%) intermediate HCC. Resection was performed in 11/12 (92%) early HCC
eligible for PT and in 1/33 (3%) advanced HCC.
Conclusions: The method of a multidisciplinary HCC-team 1)
seems to overcome some limitations due to local unavailability of
liver surgery, as shown by high rates of adherence to qualified recommendations, with well-founded reasons for discrepancies and
for referring to liver surgery centres; 2) provides the advantages of
a collegial choice and often reduces useless/costly patient mobility.
e26
T-16
T-17
POST-TRANSLATIONAL REGULATION OF
POLYCYSTIN 2 (PC2)EXPRESSION AS A NOVEL
MECHANISM OF CHOLANGIOCYTE REACTION TO
BILIARY DAMAGE AND REPAIR
ANTIVIRAL THERAPY WITH SOFOSBUVIR PLUS
RIBAVIRIN FOR THE TREATMENT OF SEVERE
HCV RECURRENCE AFTER LIVER
TRANSPLANTATION: PRELIMINARY DATA FROM
A SINGLE-CENTRE EXPERIENCE
C. Spirli 1,∗ , A. Villani 1 , C.M. Morell 3 , L. Fabris 2 ,
R. Fiorotto 1 , M. Strazzabosco 1,3
1 Section of Digestive Diseases, Yale University, New
Haven, Connecticut, USA
Padua, Padua, Italy
3 Department of Surgery and Interdisciplinary
Medicine, University of Milan-Bicocca, Milan, Italy
Introduction: In polycystic liver diseases, defective PC2 leads
to elevated production of cAMP, PKA-dependent activation of
the ERK1/2 pathway, HIF1␣-mediated VEGF production and
stimulation of cyst growthand progression. Activation of the
ERK/HIF1␣/VEGF pathway in cholangiocytes is fundamental during repair from biliary damage. In this study we hypothesized that
PC-2 levels can be modulated during biliary damage/repair.
Results: PC2 protein expression was significantly reduced in
livers from mice undergoing cholangiocyte damage (Mdr2-/- -KO,
BDL, treatment with dehydrocollidine–DDC). However, PC-2-gene
expression was not reduced,suggesting that the decrease in PC2
was due to increased degradation. To understand if factors involved
in biliary damage influence PC2 protein expression, mouse cholangiocytes were treated with pro-inflammatory cytokines, nitric
oxide (NO) donors and ER stressors. Expression of PC2 protein
was again significantly reduced, but not its gene expression.
Noteworthy, downregulation of PC-2 leads to increased ERK1/2
phosphorylation, HIF1␣ transcription activity and secretion of
VEGF. Expression of Herp and NEK, proteins that promote PC2
degradation via the proteasome complex was also increased.
Pre-treatment with proteasome inhibitor restored the expression
of PC2 in cells treated with cytokines but not in cells treated
with NO donors or with ER stressors. In the latter conditions,
PC2 degradation was inhibited by inhibition of PI3 K, that blocks
autophagosome and by blockade of lysosomes, suggesting a role for
autophagy. Finally, treatment of DDC-treated mice with the proteasome inhibitor bortezomib, significantly reduced the extent of the
liver damage and restored PC2 expression.
Conclusion: PC2 is modulated post-translationally by proinflammatory cytokines, ER-stressors and NO-donors and
is reduced in mice with biliary damage. Down regulation
of PC2 protein expression in cholangiocytes increases the
ERK1/2/HIF1␣/VEGFsecretion, thereby playing a pivotal role
in the regulation of cholangiocyte response to biliary damage.Treatments able to restore PC2 expression may represent a
new therapeutic approach in biliary diseases.
D. Arese 1,∗ , S. Martini 1 , S. Strona 1 , M. Sacco 1 ,
D. Cocchis 2 , S. Mirabella 2 , G. Rizza 2 , F. Tandoi 2 ,
R. Romagnoli 2 , A. Ottobrelli 1 ,
M.R. Torrani-Cerenzia 1 , F. Balzola 1 , M. Salizzoni 2 ,
M. Rizzetto 1
1 Gastrohepatology Unit, A.O.U. Città della Salute e
della Scienza di Torino, Torino, Italy
2 Liver Transplant Centre, General Surgery Unit,
A.O.U. Città della Salute e della Scienza di Torino,
Torino, Italy
Background and aims: Recurrent hepatitis occurs in the majority
of HCV viremic recipients at liver transplantation (LT). We evaluated safety and efficacy of antiviral therapy (Tx) with sofosbuvir
(SOF) plus ribavirin (RBV) in a population of patients (pts) having
severe recurrent hepatitis after LT.
Methods: Among HCV viremic pts transplanted in our Centre
since 2000, we enrolled to date 107 pts affected by HCVRNApositive recurrent hepatitis with fibrosis score F3 (21 pts) or F4 (86
pts) according to Metavir. 78% male, mean age 60.5 years, mean
BMI 24.7; 49% receiving cyclosporine, 43% mycophenolate, 38%
tacrolimus and 3% m-TOR inhibitors. GT1 75% (GT1a 13%), GT2 5%,
GT3 15%, GT4 5%; IL28CC 24%; 65% experienced. According to compassionate use, pts receive SOF 400 mg/die plus RBV (weight-based)
for 24 weeks. Median time from LT to Tx 4.96 years (0.25-14.25);
mean baseline HCVRNA 6.62 log10 IU/mL (3.4-7.9 log10 IU/mL);
mean GFR 76 mL/min.
Results: Until now, 31/107 pts completed 12 weeks of Tx. After
1 week of Tx, median decrease in HCVRNA was 3.61 log10 IU/mL;
50/98 (51%) at week 4, 76/77 (99%) week 8, 31/31 week 12, 9/9
week 16, 2/2 week 24 on Tx were HCVRNA negative. Of the 2
pts who completed the planned therapy and reached 4 weeks
post-Tx, one relapsed; he is IL28TT, GT1a and the only one who
tested HCVRNA < 15 IU/mL at w8 on Tx. The most common adverse
events were fatigue, headache and nausea. During Tx, 1 patient had
variceal bleeding (day 78) and 1 died due to multiorgan failure (day
86); 25% received blood transfusion/epoetin. Minimal immunosuppression dose adjustments were required on Tx and no rejections
were recorded.
Conclusions: In pts with severe HCV recurrence post-LT, antiviral regimen with SOF plus RBV is well tolerated and easy to manage,
allowing rapid HCV clearance.
T-18
T-19
ASSESSMENT OF FREE LIGHT CHAINS IN HCV
POSITIVE PATIENTS WITH MIXED
CRYOGLOBULINEMIA VASCULITIS UNDERGOING
RITUXIMAB TREATMENT
PNPLA3 RS738409 I748 M IS ASSOCIATED WITH
STEATOHEPATITIS IN NON OBESE SUBJECTS
WITH HEPATITIS C
Basile 1,∗ ,
Gragnani 2 ,
Torti 1 ,
Piluso 2 ,
U.
L.
E.
A.
F. Gulli 1 , T. Urraro 2 , M.T. Dell’Abate 1 , C. Stasi 2 ,
M. Monti 2 , G.L. Rapaccini 3 , A.L. Zignego 2
1
Department of Laboratory Medicine of the Catholic
University of the Sacred Heart, Rome, Italy
2 Department of Experimental and Clinical Medicine,
3 Institute of Internal Medicine, Catholic University
of the Sacred Heart, Rome, Italy
Introduction: Mixed Cryoglobulinemia (MC) is an HCV-related
lymphoproliferative disorder, secondary to a systemic vasculitis of
small vessels. Treatment with anti-CD20 monoclonal antibodies
Rituximab (RTX) is proved to be very useful. Free light chain (FLC)
␬/␭ ratio and FLC patterns were associated with MC and/or B-non
Hodgkin’s lymphoma.
Aims: The aim of this study was to evaluate changes in serum
free light chains (FLC) in HCV positive patients with related Mixed
Cryoglobulinemia (MC) undergoing anti-CD20 monoclonal antibody Rituximab (RTX) therapy. Furthermore, we attempted to
correlate FLC values with therapy response.
Patients and Methods: We retrospectively enrolled 46 patients
with HCV infection (26 females, 20 males), including 10 patients
without signs/symptoms of MC-related vasculitis, 36 with MCvasculitis. Clinical and biological data were recorded at baseline
and six months after RTX treatment. Nephelometric measurement
of serum FLCs was performed using a serum FLC assay.
Results: The mean serum FLC-k level was significantly higher in
MC patients, compared to HCV patients without MC and to blood
donors (p = 0.05 and p < 0.0001, respectively); the mean serum FLCratio was significantly higher in MC patients, compared to HCV
patients without MC and to blood donors (p = 0.0023 and p = 0.0008,
respectively). An abnormal FLC-ratio at baseline correlated with
presence of cryoglobulins, C4 consumption, higher RF level and
higher vasculitis rate (p < 0.005 for each parameter).
In order to evaluate the predictive value of FLC patterns, MC
patients were divided into two groups according to RTX therapy
outcome (responders and no/partial responders).
Abnormal baseline FLC-ratio was significantly associated with
no/partial response (OR 4.86–95% C.I. 0.89-28.72, p = 0.0314).
Conclusions: RTX-treatment in HCV-related MC induces a
reduction of FLC-k and RF levels. Moreover, pre-treatment FLCratio, which can be easily assessed by a routine test, may be useful
to predict response to this expensive treatment for HCV-related MC
patients ineligible to IFN-based therapy.
e27
S. Petta 1,∗ , E. Vanni 2 , E. Bugianesi 2 , C. Rosso 2 ,
D. Cabibi 3 , C. Cammà 1 , V. Di Marco 1 , M. Eslam 4 ,
S. Grimaudo 1 , F.S. Macaluso 1 , D. McLeod 5 ,
R.M. Pipitone 1 , M.L. Abate 2 , A. Smedile 2 ,
J. George 4 , A. Craxì 1
1 Section of Gastroenterology, Di.Bi.M.I.S., University
of Palermo, Palermo, Italy
2 Division of Gastro-Hepatology, Department of
Medical Sciences, San Giovanni Battista Hospital,
3 Cattedra di Anatomia Patologica, University of
4 Storr Liver Unit
5 Institute of Clinical Pathology and Medical
Research, Westmead Millennium Institute and
Westmead Hospital, University of Sydney, Sydney,
Australia
Background and Aims: The PNPLA3/Adiponutrin rs738409 C/G
single nucleotide polymorphism is associated with the severity of
steatosis, steatohepatitis and fibrosis in patients with non-alcoholic
fatty liver disease, as well as with the severity of steatosis and fibrosis in patients with chronic hepatitis C (CHC). We tested in genotype
1 (G1) - CHC patients the putative association between the PNPLA3
variant and histological features of steatohepatitis, as well as their
impact on the severity of fibrosis.
Methods: 434 consecutively biopsied Caucasian G1-CHC
patients were genotyped for PNPLA3 rs738409. Histological features of steatohepatitis in CHC were assessed using the Bedossa
classification. Hepatic expression of PNPLA3 mRNA was evaluated
in 63 patients.
Results: The prevalence of steatohepatitis increased from 16.5%
in patients with PNPLA3 CC, to 23.2% in CG and 29.2% in GG genotype (p = 0.02). By multiple logistic regression, PNPLA3 genotype
(OR 1.54, 95%CI 1.03-2.30, p = 0.03) together with age (OR 1.03,
95%CI 1.00-1.05, p = 0.02), BMI ≥ 30 (OR 2.06, 95%CI 1.04-4.10,
p = 0.03) and HOMA (OR 1.18, 95%CI 1.04-1.32, p = 0.006) were
independently linked to steatohepatitis. When stratifying for obesity, PNPLA3 was associated with NASH in non obese patients
only (12.0% in CC vs 18.3% in CG vs 27.3% in GG, p = 0.01), including after correction for metabolic confounders (OR 2.06, 95%CI
1.26-3.36, p = 0.004). We confirmed the independent association
of rs738409 with the severity of steatosis (OR 1.71, 95%CI 1.202.45, p = 0.003), and, indirectly by steatohepatitis promotion (OR
2.05, 95%CI 1.05-4.02, p = 0.003), with severe fibrosis. Higher liver
PNPLA3 mRNA was associated both with the severity of steatosis
(adjusted p = 0.03) and steatohepatitis after adjusting for gender,
age, BMI and HOMA (p = 0.002).
Conclusions: In G1-CHC patients, the PNPLA3 G variant is associated with a higher risk of steatosis severity and steatohepatitis in
chronic hepatitis C, particularly among non obese subjects.
e28
T-20
T-21
HLA DPB1 RS9277535 POLYMORPHISM
STRONGLY PREDICTS HBSAG CLEARANCE IN
INTERFERON TREATED GENOTYPE D
HBEAG-NEGATIVE PATIENTS WITH CHRONIC
HEPATITIS B
ADD-ON PEGINTERFERON ALFA-2A
SIGNIFICANTLY REDUCES HBSAG LEVELS IN
CHRONIC HEPATITIS B, HBEAG-NEGATIVE,
GENOTYPE D PATIENTS FULLY SUPPRESSED ON
NUCLEOT(S)IDE ANALOGUES TREATMENT:
HERMES STUDY INTERIM ANALYSIS
P. Lampertico 1,∗ , E. Galmozzi 1 , F. Facchetti 1 ,
C. Cheroni 2 , F. Invernizzi 1 , V. Valveri 2 ,
R. Soffredini 1 , M. Viganò 3 , S. Abrignani 2 ,
R. De Francesco 2 , M. Colombo 1
2 INGM Istituto Nazionale Genetica Molecolare
Milan, Italy
3 Division of Hepatology, Ospedale San Giuseppe,
University of Milan, Milan, Italy
Introduction and Aim: HLA DP polymorphisms have recently
been associated to spontaneous hepatitis B virus (HBV) clearance in
Asians patients. Whether these SNPs influence IFN-induced HBsAg
response in difficult to cure HBeAg-negative genotype D chronic
hepatitis B Caucasian patients, is currently unknown.
Materials and Methods, Results: 126 such patients with compensated disease (46 years, 82% males, 90% genotype D, HBV DNA
6.2 log cp/ml, ALT 132 IU/L, 40% with cirrhosis) were followed
for a median of 11 (1-23) years after 22 (6-48) months of either
standard or pegylated (Peg)IFN alfa treatment. The primary endpoint was HBsAg clearance. HLA DPA1 rs3077, HLA DPB1 rs9277535
and rs9277534, and IL28B rs12979860 genotypes were assessed
by using TaqMan SNP Genotyping Assays. Twenty-eight (22%)
patients ultimately cleared serum HBsAg with a 15-year cumulative probability of 30%, with a preference for patients with baseline
lower HBV DNA levels, higher ALT levels, IL28B rs12979860 CC,
HLA DPB1 rs9277534 AG/GG and HLA DPB1 rs9277535 AG/GG.
At multivariate analysis, HLA DPB1 rs9277535 AG/GG genotype
was the strongest independent predictor of HBsAg seroclearance
(HR: 6.88; 95%CI 2.82-16-7, p < 0.000) along with baseline ALT levels (HR 1.00, 95%CI 1.00-1.00, p = 0.001), baseline HBV DNA levels
(HR 0.54, 95%CI 0.37-0.79, p = 0.002), and IL28B rs12979860 CC
genotype (HR 2.2, 95%CI 1.02-5.0, p = 0.044). The 15-year cumulative rates of HBsAg clearance were 62% in carriers of the HLA
DPB1 rs9277535 AG/GG genotype compared to 13% in carriers of
the AA genotype (p < 0.0000). Conclusions: HLA DPB1 rs9277535
polymorphism strongly predicts IFN-induced HBsAg clearance in
HBeAg-negative patients chronically infected by genotype D HBV.
This genetic signature may help to select patients to IFN based
regimen.
P. Lampertico 1,∗ , M.R. Brunetto 2 , A. Craxì 3 ,
G.B. Gaeta 4 , M. Rizzetto 5 , G. Palmieri 6 ,
M. Colombo 1
Ospedale Maggiore Policlinico, University of Milan,
Milan, Italy
2 Liver Unit, Reference Centre for Chronic Liver
Desease and HCC of the Tuscany Region, University
Hospital of Pisa, Pisa, Italy
3 Gastroenterology and Hepatology, Di.Bi.M.I.S.,
University of Palermo, Palermo, Italy
4 Department of Internal Medicine, Section of
Infectious Desease, Second University of Naples,
Naples, Italy
5 Department of Gastroenterology, University of
Turin, Turin, Italy
6 Roche S.p.A., Monza, Italy
Backgound: Since nucleos(t)ide Analogues (NA) suppress viral
replication in HBeAg-negative CHB without inducing significantly
HBsAg loss, several studies are investigating the impact to HBsAg
levels of Peginterferon alfa-2a (PEG-IFN) addition to NA treated
patients. Here we present the week 24 analysis of an ongoing trial
evaluating PEG-IFN add-on in NA treated CHB, HBeAg-negative,
genotype D, Caucasians patients (HERMES study). Methods: In this
phase IIb, open label, single arm, multicenter Italian study, patients
who were receiving NA monotherapy, with HBV-DNA persistently
below 20 IU/ml for at least 12 months and HBsAg > 100 IU/ml, initiated add-on treatment with Peginterferon alfa-2a 180 ␮g sc once
weekly for 48 weeks. Patients without any HBsAg decrease at week
24 or those who will complete the add-on treatment period (week
48) are going to discontinue PEG-IFN and be followed-up for additional 48-weeks. Serum HBsAg decline at the end of treatment
with PEG-IFN plus NA combination (week 48) is the primary study
endpoint. Results: 70 Caucasians who had completed 24 weeks of
combination treatment were included in this analysis. Median age
was 50.5 (29-64) and 81% were males. At week 24 of combination treatment, 27.1% decreased serum HBsAg≥50% from baseline,
while 15.7% discontinued study due to lack of response. Median
qHBsAg significantly decreased from 1160 IU/ml at baseline to
743 IU/ml at week 24 (p < 0.0001). The proportion of patients with
HBsAg < 500 or < 1000 IU/ml increased from baseline (23% and 47%
respectively) up to week 24 (39% and 59%, respectively). Fifty-seven
patients (81.4%) experienced adverse events (AEs), 4.3% and 5.7% of
the patients discontinued or temporarily interrupted PEG-IFN due
to AEs respectively, while in 8.6% of patients PEG-IFN dosage was
adjusted due to AEs. Conclusion: In HBeAg-negative, CHB, genotype
D patients treated with nucleot(s)ide analogues, add-on treatment
with PEG-IFN results in significant reduction of serum HBsAg levels.
T-22
T-23
ROLE OF KILLER CELL IMMUNOGLOBULIN-LIKE
RECEPTORS AND THEIR HLA CLASS I LIGANDS IN
AUTOIMMUNE HEPATITIS
SLOW ACHIEVEMENT OF HCV-RNA
UNDETECTABILITY IN CIRRHOTIC PATIENTS
TREATED WITH SOFOSBUVIR + RIBAVIRIN:
POSSIBLE CLINICAL IMPLICATIONS IN THE LIVER
TRANSPLANT LIST MANAGEMENT
R. Littera 1 , C. Carcassi 1 , L. Secci 2 , S. Lai 1 ,
L. Cappai 1 , R. Porcella 1 , F. Alba 1 , R. Maddi 1 ,
M. Serra 1 , S. Cappellini 2 , C. Salustro 2 , S. Onali 2 ,
C. Balestrieri 2 , G. Serra 2 , M. Conti 2 , T. Zolfino 3 ,
R. Scioscia 2 , L. Barca 2 , L. Chessa 2
1 Medical Genetics, Department of Medical Sciences
“M.Aresu”, University of Cagliari, Italy
2 Center for the Study of Liver Diseases, Department
of Medical Sciences “M. Aresu”, University of Cagliari,
Italy
3 S.C. Gastroenterologia, AOB, Cagliari, Italy
Introduction: It is well known that CD4, and CD8 T lymphocytes
are critical protagonists of the immunopathogenetic mechanisms
of autoimmune hepatitis (AIH). However, so far, data on the
involvement and role of natural killer (NK) cells are scarce.
Aim: Killer cell immunoglobulin receptors (KIRs) represent the
major family of cell surface receptors that inhibit and activate NK
cells. The elevated expression of NK cells in liver tissue prompted
us to evaluate the impact of NK cells and KIRs on age of onset and
evolution of the disease.
Materials and Methods: We retrospectively analyzed a total
of 114 Sardinian patients diagnosed with type I AIH. Metabolic
and genetic causes of hepatopathy were excluded in all patients.
Patients and controls were typed at high resolution (4 digits) for
the alleles at the HLA-A, -B, -C and DR loci and at 15 KIR gene
loci. Patients were divided into 2 groups according to homozygosity
for KIR haplotype A (KIR genotype AA), heterozygosity or homozygosity for KIR haplotype B (KIR genotype Bx).The immunogenetic
characteristics of the AIH patients were compared with those of
221 healthy individuals from the Sardinian bone marrow donor
registry.
Results: The activating KIR gene KIR2DS1 had a significantly
higher frequency in AIH patients compared to controls [57% vs
43%; HR (95% CI) = 1.7 (1.0-2.7); P = 0.03]. Also the presence of
KIR2DL1 + /KIR2DS1 + /HLA-C2 + was higher in AIH patients [47.3%
vs 35.7%; HR (95% CI) = 1.6 (1.0-2.6); P = 0.04].
Conclusions: Based on our findings, it can be hypothesized that
NK cells are involved in the pathogenetic mechanisms of type I AIH
and more specifically, that the activating KIR gene KIR2DS1 is associated with the development of the disorder. The role of KIR2DS1
as a marker for AIH warrants further investigation.
e29
I. Lenci 1 , V. Cento 2 , M. Rendina 3 , M.F. Donato 4 ,
M. Milana 1 , D. Sforza 5 , M. Manuelli 5 , M. Aragri 2 ,
V.C. Di Maio 2 , A. Abedrabbo 1 , A. Castellaneta 3 ,
F. Malinverno 4 , S. Monico 4 , M.L. Ponti 6 ,
R. Canu 6 , R. Ganga 6 , R. Alfieri 7 , L. Milanesi 7 ,
A. Di Leo 3 , G. Tisone 5 , C.F. Perno 2,8 ,
F. Ceccherini-Silberstein 2 , M. Colombo 4 ,
M. Angelico 1
1 Hepatology Unit, Policlinico Tor Vergata, Tor
Vergata University, Rome
2 Virology Chair, Tor Vergata University, Rome
3 Gastroenterology and Digestive Endoscopy,
University Hospital, Policlinico Bari
4 Gastroenterology and Liver Transplant Units, Cà
Granda Maggiore Hospital Policlinico and University
of Milan, Milan, Italy
5 Liver Transplant Centre, Policlinico Tor Vergata, Tor
Vergata University, Rome
6 Grastroenterology and Hepatology Units, Brotzu
Hospital, Cagliari, Italy
7 Institute for Biomedical Technologies-CNR, Segrate,
(Milano)
8 Virology Unit, Policlinico Tor Vergata, Tor Vergata
University, Rome
Sofosbuvir (SOF) treatment ± ribavirin (RBV) prior to LT has the
potential to change the HCV recurrence after Liver Transplantation
(LT). This approach has been reported to avoid graft reinfection
in compensated patients with hepatocellular carcinoma (HCC),
but only among those who reached and maintained undetectable
HCV-RNA (TND) before LT. Since the time to obtain this result in
decompensated cirrhotics is unknown, we sought to investigate
the early HCV-RNA decay in this setting.
Sixteen decompensated patients (M/F 12/4, median age 55.3,
CPT score ≥ B7), infected by HCV genotype 1a, 1b, 3 and 4 (2-84-2), 4 of whom with HCC, were treated with SOF 400 mg/day
and RBV (200-1000 mg/day), except 2 who received SOF alone, for
a median(IQR) of 12(11-16) weeks awaiting LT. HCV-RNA levels
were measured weekly and safety and clinical parameters were
analyzed.
No serious adverse events were reported. The median(IQR)
RBV dose was 600(400-800). Despite 11/16 patients had low
baseline viremia (<600.000 IU/ml), HCV-RNA decay in the first 4
weeks of treatment was suboptimal (median[IQR] = -3.7[-4.3;-3.3]
LogIU/ml). Only 3/16 (18.7%) patients reached TND HCV-RNA at
week 4 (rapid virological response, RVR), and 5/10 (50%) evaluable patients were still viremic at week-12. Median(IQR) MELD
decreased from 15(13-16) at baseline to 13(12-15) at week 4,
when 14/16 (87.5%) patients returned in a compensated stage. One
patient underwent LT after 6 weeks of treatment, while HCV-RNA
was still positive (26 UI/mL). SOF was interrupted only during the
e30
first 4 perioperative days, obtaining TND HCV-RNA at week 10 of
therapy.
The kinetics of HCV-RNA decay in decompensated cirrhotic
patients are slower compared to those obtained in non cirrhotic
patients. As a practical implication, pre-LT treatment may need to
be longer and/or based on more effective antiviral strategies. Alternatively, MELD-based prioritization to LT should be reassessed in
light of the results of antiviral therapy.
T-24
CHRONIC HEPATITIS B TREATMENT
INDIVIDUALIZATION BY MEANS OF SERUM
HBSAG AND MIR-B-INDEX KINETICS
Cavallone 1 ,
Oliveri 1 ,
Colombatto 1 ,
D.
F.
P.
B. Coco 1 , P. Ciccorossi 1 , V. Romagnoli 1 ,
B. Cherubini 1 , F. Moriconi 1 , F. Bonino 2 ,
M.R. Brunetto 1
1
Hepatology Unit, Reference Center of the Tuscany
Region for Chronic Liver Disease and Cancer,
University Hospital of Pisa, Italy
2 Digestive and Liver Disease, General Medicine II
Unit, University Hospital of Pisa, Italy
Background and Aims Non-viral biomarkers of sustained
control of HBV infection are an unmet need for treatment individualization in Chronic-Hepatitis-B (CHB). We reported that
Inactive-Carriers and patients with sustained-virologic-response
(SVR) to Peg-IFN share a common serum miRNA signature (MiR-BIndex, MBI). We studied the kinetics of MBI and HBsAg during/after
Peg-IFN.
Patients and Methods 46 CHB patients (11 HBeAg-pos, 29
males, median-age 48.6y, 22.4-64.1y; genotypes: A 5, D 39, F 2)
were treated with Peg-IFN2a 180 ␮g/w (median 13.2months, 824 m): 20 SVR; 17 relapsers (REL) and 9 non-responders (NR).
The quantitative analysis of miR-122-5p, miR-99a-5p, miR-192-5p,
miR-126-3p, miR-335-5p and miR-320a was performed as previously reported. HBsAg and MBI were tested at baseline (BL),
24-48w, end-of-therapy (EOT), 24w-post-treatment (PTFU).
Results MBI > -1.7 were found: at BL in 5 HBeAg-neg pts (4 SVR,
1 NR); at EOT in 14/19 (73.7%) SVR (4 HBeAg-pos, 10 HBeAg-neg)
and at PTFU in all SVR, but in 1 NR and 1 REL. Kinetics of HBsAg
and MBI in HBeAg-neg pts are reported in the table; MBI≥-3.0
and HBsAg < 1000 IU/mL at EOT had: 100-93.3% sensitivity, 94.785.0% specificity, 93.3-82.4% PPV, 100-94.4% NPV, 97-88.6% DA in
identification of SVR (AUROC 0.988 and 0.951 respectively).
Conclusions Overall HBsAg and MBI show good performances
in SVR prediction. In HBeAg-neg-CHB MBI predicts SVR since therapy beginning and identifies all SVR at EOT qualifying as the best
biomarker for individual therapy monitoring.
SVR(15)
REL(15)
NR(5)
P*
T-25
HBX–DLEU2 LNCRNA COMPLEX AFFECTS
TRANSCRIPTION OF NEW TARGET PROMOTERS
F. Guerrieri 1,2 , L. Chiodo 1 , S. Jeddari 2 ,
D. D’Andrea 3 , A. Tramontano 3 , G. Ruocco 1 ,
M. Levrero 1,2
1 CLNS@SAPIENZA, Istituto Italiano di Tecnologia
(IIT), Rome, Italy
2 Department of Internal Medicine, Sapienza
3 Biocomputing Lab, Department of Physics, Sapienza
Background: A ChIPSeq analysis of HBx genome wide recruitment identified 39 long non coding RNAs (lncRNAs), including
DLEU2, as direct HBx transcriptional targets in HBV replicating
cells. DLEU2 lncRNA overlaps the first exon of the TRIM13 gene and
the pri-Mir of the miR-15a-16.1 cluster. Up-regulation of specific
DLEU2 splicing variants correlates with tumors.
Objectives: Aim of this study was to investigate HBx role in
DLEU2-TRIM13-mir15.mir16 regulation.
Methods: Anti-HBx ChIP were performed in HepG2 cells replicating wt and HBx-mt HBV. HBx ChIPed DNA and lncRNA-gene
expression levels were analysed by TaqMan RT-PCR using specific
primers. Specific LNATM longRNA GapmeRs (Exiqon) were used for
highly efficient inhibition of DLEU2 lncRNA function. The I-Tasser,
RNAfold and 3dRNA softwares were used for prediction of HBx protein structureand RNA secondary and tertiary structures.
Results: HBx binding to the DLEU2 promoter leads to increased
H4 acetylation, a different DLEU2 splicing profile, down-regulation
of miR-15a-16.1 cluster and up-regulation of the antisense
autophagic gene TRIM13, in the absence of HBx binding to the
TRIM13 promoter. GapmeR-mediated selective degradation of
DLEU2 RNA results in reduced TRIM13 promoter H4 acetylation
and a ∼50% reduction of TRIM13 expression in HBV replicating
cells. These results directly link DLEU2 RNA species with TRIM13
transcriptional regulation in the presence of HBx. The interaction
between HBx and DLEU2 was confirmed by RIP (RNA Immune
Precipitation) experiments. By coupling ab initio modeling of HBx
protein and DLEU RNA tertiary structures and we constructed a
docking model of DLEU2-HBV complex. Finally, we found that
DLEU2 inactivation has a profound impact on pgRNA transcription, suggesting a functional relevance of DLEU2-HBx interaction
for HBV replication.
Conclusion: HBx targets the DLEU2 promoter leading to a different DLEU2 splicing profile and its binding to DLEU2 RNA species
affects cellular genes transcription and HBV replication.
HBsAg (Log10 IU/mL)
BL
T24w
T48w
EOT
PTFU
P**
3.45
3.76
3.70
.042
2.81
3.63
3.60
.001
2.69
3.50
3.43
< .001
2.15
3.44
3.58
< .001
1.33
3.34
3.10
< .001
.002
.008
.607
*Mann-Whitney (SVR vs REL + NR); **ANOVA (BL vs EOT).
MiR-B-Index
BL
T24w
T48w
EOT
PTFU
P**
−4.49
−9.81
−7.26
.001
−1.65
−8.31
−8.29
< .001
1.16
−8.80
−7.20
< .001
1.46
−7.22
−8.82
< .001
4.60
−4.90
−5.92
< .001
< .001
.034
.335
T-26
T-27
IMPULSIONAL, POINT AND BIDIMENSIONAL
SHARE WAVE ELASTOMETRY FOR PORTAL
HYPERTENSION: SAME STIFFNESS THRESHOLD?
SERPINB3 AND YAP INTERPLAY INCREASES MYC
ONCOGENIC ACTIVITY
H. Stefanescu, G. Allegretti, C. Serra, G. Marasco,
N. Gamal, F. Conti, A. Colecchia, D. Festi,
P. Andreone, L. Bolondi, F. Piscaglia
S.Orsola-Malpighi Hospital, Alma Mater Studiorum University of Bologna, Bologna, Italy
BACKGROUND: Liver stiffness (LS) is used to investigate the
presence of portal hypertension (PH). LS measured with Transient
elastography (TE) correlates with HVPG, and 21.1 kPa is proposed
as cut-off to predict clinically significant PH (CSPH). New elastographic techniques may overcome some limitations of TE, but their
diagnostic accuracy remain to be defined. We aimed to compare
the performance of TE, point shear wave elastography (ElastPQ)
and 2D-Shear Wave Elastography (2D SWE) to predict the grade of
PH in a cohort of cirrhotic patients.
METHODS: 65 consecutive patients (63.1%M, 60.2y) with
chronic liver disease submitted to HVPG measurement and at
least one elastographic technique: TE and/or ElastPQ (Philips IU22)
and/or 2D SWE (Aixplorer by Supersonic Imagine) blindly one from
each-other, were enrolled between December 2013 and November
2014.
RESULTS: 44/65 patients (67.7%) had CSPH (HVPG > 10 mmHg).
40 patients (61.5%) underwent TE, 31(47.7%) ElastPQ, 25(38.5%) 2DSWE measuring liver stiffness.
HVPG was well correlated with either TE (r = 0.695;p < 0.0001),
ElastPQ (r = 0.684;p = 0.001), or 2D-SWE (r = 0.762;p < 0.0001).
The overall concordance between elastographic techniques
(tested in 51 patients who underwent either 2 or 3 methods) was
only moderate (ICC = 0.409). TE was concordant with both other
techniques (ICC = 0.772 for ElastPQ, ICC = 0.754 for 2D-SWE), while
ElastPQ and 2D-SWE were less concordant (ICC = 0.488).
No significant difference was detected among AUROCs for predicting CSPH (0.872 for TE, 0.962 for ElastPQ and 0.860 for 2D-SWE,
respectively).
For the 21.1 kPa cutoff, the accuracy of each technique to correctly classify CSPH was: 85%(34/40) for TE, 55%(17/31) for ElastPQ
and 76%(19/25) for 2D-SWE, respectively.
CONCLUSION: Our findings showed that liver stiffness, assessed
by new elastographic techniques (ElastPQ and 2D-SWE), have
similar performance to predict CSPH like TE. However the best
diagnostic threshold for this diagnosis differs significantly among
different elastography technologies and specific diagnostic accuracy studies appear warranted for each type of equipment.
e31
C. Turato 1 , S. Cannito 2 , D. Simonato 1 ,
G. Villano 1,2,3 , L. Terrin 1 , S. Quarta 1 , A. Biasiolo 1 ,
M. Ruvoletto 1 , S. Fasolato 3 , G. Zanus 3 , U. Cillo 3 ,
A. Gatta 1 , M. Parola 2 , P. Pontisso 1
1 Department of. Medicine, University of Padua,
Padua, Italy
2 Department of. Clinical and Biological Sciences,
3 Department of Surgical, Oncological and
Gastroenterological Sciences, University of Padua,
Padua, Italy
Background SerpinB3 has been recently described as an early
marker of liver carcinogenesis. Myc is one of the most important
oncogenes in human cancer. Somatic amplification and overexpression of Myc often correlate with more advanced and aggressive
tumour forms. Yes-associated protein (Yap), the main effector of
the Hyppo pathway, is a central regulator of proliferation and
self-renewal of normal and cancer stem cells. This molecule,
found up-regulated in hepatocellular carcinoma (HCC), has been
described also to increase Myc expression.
Aim The present study has been designed in order to investigate whether SerpinB3 may functionally modulate Myc in different
experimental models and in human HCC specimens.
Materials and Methods Expression of Myc, Yap and its target
genes was evaluated in relation to SerpinB3 expression in HCC
specimens, in C57BL/6 mice transgenic for human SerpinB3 and
in hepatoma cells transfected with plasmid carrying the SerpinB3
gene. Moreover the inhibitory effect of SerpinB3 on calpain was
assessed by means of the calpain activity assay.
Results A positive correlation between Myc and SerpinB3
expression was observed at transcription and protein level in HCC
specimens, where Myc oncogene was found predominantly in the
nucleus of cancer cells overexpressing SerpinB3. Myc expression
was significantly up-regulated by SerpinB3 through calpain and
Hyppo-dependent molecular mechanisms. Recombinant SerpinB3
protein was indeed capable to inhibit the activity of calpain in
vitro, likely reducing its ability to cleave Myc in its non-oncogenic
Myc-nick cytoplasmic form. Furthermore, SerpinB3 increased the
transcription of Myc through the induction of Yap pathway, as
documented by a remarkable Yap nuclear translocation and upregulation of Yap target genes.
Conclusions Data from the present study provide evidence that
SerpinB3 can improve the production of Myc oncogene through
direct and indirect mechanisms that include the inhibition of generation of its cytoplasmic form and the activation of Yap pathway.
e32
T-28
T-29
TRANSARTERIAL CHEMOOCCLUSION (TACO)
WITH DEGRADABLE-STARCH-MICROSPHERE IN
UNRESECABLE HEPATOCELLULAR CARCINOMA:
A PROSPECTIVE PILOT-STUDY
HIF2␣ NEDDYLATION AS A SELECTIVE
SERPINB3-DEPENDENT MECHANISM LEADING
TO ITS INCREASED STABILIZATION AND
NUCLEAR TRANSLOCATION IN LIVER CANCER
CELLS
S. Francioso, A. Orlacchio, F. Chegai, F. Santopaolo,
I. Lenci, M. Milana, A. Brega, M. Angelico
Liver and Transplant Unit, Tor Vergata University,
Rome, Italy
Background: According to BCLC algorithm, Transarterial
Chemoembolization is recommended for unresecable hepatocellular carcinoma (HCC) in patients with preserved liver function. The
intraarterial chemotherapy with transient artery occlusion could
be a valid treatment alternative in patients with high-risk of liver
failure.
Aim of the present study was to evaluate efficacy and safety
of TACO using Degradable-Starch-Microspheres (DSM) in patients
with unresecable HCC.
Methods We prospectively enrolled 24 cirrhotic patients
(21/3 M/F, mean age 66.3 ± 10.5 years) with unresecable HCC, to
be treated with three consecutives DSM-TACO at 5-week intervals. Chemotherapy drug was Doxorubicin Chloridrate (50 mg/m2 )
mixed with 275 mg DSM (Embocept®S, PharmaCept). Liver function before and after treatments was evaluated with Child Pugh and
MELD scores. Treatment response was assessed by CT-scan 4 weeks
after procedures according to mRECIST criteria.
Results: Overall, Complete Response (CR) was observed in 5/24
(20.8%), 9/24 (37.5%) and 14/24 (58.3%) pts after the first, the second
and the third procedure, respectively. No patient showed Stable
Disease or Disease Progression at the end of the study. Patients
with monolobar disease (14/24: 58.3%) showed significant CR after
the 1st procedure compared with bilobar HCC localization (5/14
vs 0/10; p = 0.017). At the end, CR did not differ between mono
or bilobar disease (9/14: 64.2% vs 5/10: 50%; p = ns). Drop-outs
occurred in 8 patients (33.3%): 4 after the 1st treatment (3 worsened liver function, 1 liver transplanted), 4 after the 2nd treatment
(2 worsened liver function, 1 liver transplanted, 1 non compliance).
Postembolization syndrome was observed in 9 (37.5%), in 4 (16.6%)
requiring extended hospitalization. One patient had cholecystitis
resolved with medical care.
Conclusions: DSM-TACO offers a valid TACE alternative in
patients with unresecable HCC. Careful patients selection is
required to avoid liver failure in patients with border-line liver
compensation. Further investigation to define DSM-TACO effects
on survival is warranted.
S. Cannito 1 , G. Villano 2 , C. Turato 2 , E. Morello 1 ,
C. Paternostro 1 , E. Novo 1 , S. Quarta 2 ,
S. Colombatto 3 , F. Lopitz-Otsoa 4 ,
M.L. Martínez-Chantar 4 , P. Pontisso 2 , M. Parola 1
1 Department of Clinical and Biological Sciences, Unit
of Experimental Medicine, University of Turin, Turin,
Italy
Padua, Italy
3 Department of Oncology, University of Turin, Turin,
Italy
4 Department of Metabolomics, CIC bioGUNE, Centro
de Investigacion Biomedica en Red de Enfermedades
Hepaticas y Digestivas (Ciberehd), Technology Park
of Bizkaia, Bizkaia, Spain
Background SerpinB3, a cysteine-proteases inhibitor overexpressed in hepatocellular carcinoma (HCC), has been reported to
be up-regulated by hypoxia through a HIF2␣-dependent mechanism and to act as a paracrine mediator able to induce stabilization
and increased nuclear translocation of HIF2␣ and up-regulation
of HIF2␣-related genes in liver cancer cells. The ubiquitin-like
molecule NEDD8 is a key regulator of cell growth, viability and
malignant transformation and neddylation promotes stabilization
of proteins with essential regulatory roles. Overexpression of global
neddylation has been detected in HCC and suggested to be associated with its poorest prognosis.
Aims To investigate whether SerpinB3 may induce selective
HiF2␣ neddylation/stabilization in liver cancer cells.
Methods The cross-talk between SerpinB3 and HIF2␣ has been
investigated by taking advantage of morphological, molecular and
cell biology techniques in the following experimental models: i)
control HepG2 exposed to recombinant SerpinB3 (rSerpinB3); ii)
HepG2 stably transfected to overexpress SerpinB3 (HepG2/SB3);
iii) transgenic mice overexpressing SerpinB3 in the liver.
Results Immunohistochemistry performed onliver sections
from SerpinB3 transgenic mice shows an impressive level of nuclear
staining for HIF2␣ and exposure of HepG2 to rSerpinB3 results in
increased HIF2␣ nuclear levels. Different experimental approaches
revealed that in relation to the action of SerpinB3: increased HIF2␣
protein levels are i) unrelated to its increased transcription as well
as ii) to an inhibition of overall proteasome activity by SerpinB3;
iii) overexpression of SerpinB3 leads to an increase in the level
of neddylated proteins and of levels of NEDD8 activating enzyme
1 (NAE-1); iv) the use of a specific inhibitor of NAE-1 selectively
reduces HIF2␣ levels.
Conclusions SerpinB3 can affect the behaviour of target cells by
increasing protein levels of HIF2␣, followed by its nuclear translocation and induced transcription of target genes, by inducing HIF2␣
selective neddylation and consequent stabilization which is independent on hypoxia.
T-30
T-31
D-DIMER AND FIBRINOLYTIC ACTIVITY IN
PATIENTS WITH DECOMPENSATED LIVER
CIRRHOSIS
SURVIVAL OF PATIENTS WITH HEPATOCELLULAR
CARCINOMA (HCC) WITHIN THE BOLOGNA LIVER
ONCOLOGY GROUP: COMPARISON WITH
INTERNATIONAL GUIDELINES
R.G. Romanelli a , A.P. Cellai b , D. Lami b ,
F. Natucci a , C. Tosti-Guerra a , R. Abbate b ,
D. Prisco b , G. Laffi a
a
Dipartimento Medicina Clinica e Sperimentale
(DMSC) Liver Unit, Italy
b Sezione delle Malattie Aterotrombotiche del
Dipartimento dell’Area Critica Medico Chirurgica
Università di Firenze - Azienda Ospedaliero
Universitaria Careggi (AOUC), Firenze, Italy
Background and Aims: Cirrhotic plasma could generate similar or even greater amount of thrombin; a procoagulant imbalance
has been introduced. Ascitic fluid has been hypothesized to be the
origin of hyperfibrinolysis. Standard tests (INR) fail to really reflect
bleeding tendency. Aim of this study was to determine whether a
fibrinolytic activity is detectable in ascites.
Material andMethods: We evaluated 33 patients with liver cirrhosis (11 in Child-Pugh class A, score 5.6, mean age 65 ± 11 (yrs);
9 in Child-Pugh class B, score 7.9, mean age 69 ± 17 (yrs); 13 in
Child-Pugh class C, score 11.4, mean age 70 ± 12 (yrs) and 21 control
healthy subjects, mean age 68 ± 14 (yrs). We studied Clot Lysis Time
(CLT), D-dimer, tissue plasminogen activator (t-PA), plasminogen
activator inhibitor (PAI-1), alpha2-antiplasmin (a2AP), plasminogen (PLG), thrombin activatable fibrinolysis inhibitor (TAFI), and
(through EuroCLOT) clot formation, structure, and lysis.
Results: We found that a2AP (%), PLG (%), TAFI (mg/mL) and
fibrinogen (mg/dl) levels were significantly higher in plasma than
ascites (p < 0.001) and lower in plasma from patients than controls (p < 0.05). Instead, D-dimer levels (ng/ml) were significantly
lower in plasma than in ascitic fluid (p < 0.001), whereas similar
concentrations were found for t-PA and PAI-1. D-dimer, PAI-1 and
t-PA levels were significantly higher (p < 0.05) and CLT shorter in
plasma from patients than from healthy subjects (p < 0.05). By EuroCLOT, statistically significant differences were observed between
cirrhotic patients and healthy controls (AUC control subjects: 585;
Child-Pugh A 316*; Child-Pugh B 257*; Child-Pugh C 129*** (from
p < 0.05* to p < 0.001***).
Conclusion: Cirrhotic patients show hyperfibrinolytic activity
versus healthy subjects. In ascitic fluid from cirrhotic patients high
levels of D-dimer were found. These data further suggest that ascitic
fluid compartment is in continuous exchange with plasmatic compartment, possibly through lymphatic flux, and contributes to the
coagulopathy of liver cirrhosis.
e33
E. Terzi, M. Piccinnu, F. Piscaglia, S. Leoni,
A. Granito, L. Bolondi, on the behalf of the
BLOG-Bologna Liver Oncology Group
Division of Internal Medicine, Department of
Medical and Surgical Sciences, University of Bologna,
Bologna, Italy
Introduction: The Barcelona Clinic Liver Cance system (BCLC),
endorsed by the latest international EASL-EORTC and AASLD guidelines, represents the reference therapeutic and prognostic staging
system.
Aim: was to evaluate the overall survival and clinical determinants of survival in our series of HCC patients and to compare
the results to those of guidelines eventually attempting to identify
reasons for discrepancy.
Materials and Methods: Among 1028 HCC patients referred to
our center between January 2000 and August 2013, we retrospectively identified the outcome of the 595 consecutive patients seen
in our center on occasion of the first diagnosis of HCC.
Results: Stage was BCLC-0 in 23 patients (4%), BCLC-A in 273
(46%,), BCLC-B in 155 (26%), BCLC-C in 114 (19%) and BCLC-D in 30
patients (4%). Median survival in BCLC-0 was 88 months (95% C.I.
41.3-134.7) with 1-, 3- and 5-year survival rates of 97%, 86% and
61%; in BCLC-A was 44 months (95% C.I. 37.4-50.4) with 1-, 3- and
5-year survival rates of 92%, 60% and 36%; in BCLC-B was 20 months
(95% C.I. 14.2-25.8) with 1-, 3- and 5-year survival rates of 65%, 34%
and 21%; in BCLC-C was 8 months (95% C.I. 5.2-10.8) with 1-, 3- and
5-year survival rates of 39%, 2% and 0%; in BCLC-D was 7 months
(95% C.I. 4.8-12.2). At multivariate survival analysis, age > 67 years,
neoplastic portal vein thrombosis, AFP > 19 ng/mL and BCLC C-D
were statistically associated with shorter survival.
Conclusion: The present study validates survival data reported
by the guidelines in a large unselected series of consecutive patients
with HCC managed under real life conditions. Such results were
achieved through a multidisciplinary and individually tailored
treatment strategy of HCC patients within the Bologna Liver Oncology Group (BLOG).
T-32
TACE WITH CONE BEAM COMPUTED
TOMOGRAPHY IS MORE EFFECTIVE THAN
TRADITIONAL TECHNIQUE IN BCLC A HCC
PATIENTS INELIGIBLE TO SURGERY
R. Patti 1,2 , N. Mezzina 1,2 , F. Masutti 1,2 ,
C. Abazia 1,2 , V. Lanzillotti 1,2 , D. Pascut 2 ,
C. Sukowati 2 , F. Pozzi Muceli 3 , C. Tiribelli 1,2 ,
S.L. Crocè 1,2
1 Dipartimento di Scienze Mediche, Clinica Patologie
del Fegato, Università di Trieste, Trieste
2 Fondazione Italiana Fegato, Italy
3 Dipartimento di Scienze Mediche, UCO di
Radiologia, Università di Trieste, Trieste
Introduction: In the last years despite the improvement in cancer diagnostic and therapeutic tools HCC mortality is still high. The
decision-making process in HCC treatment is mainly based on the
e34
Barcelona Clinic Liver Cancer (BCLC) staging system that consider
both clinical and tumor characteristics. BCLC A patients should
undergo surgical ablation but they are often not eligible due to
comorbidities or tumor position. So the only treatment available
is the trans-arterial-chemoembolization (TACE) performed either
with traditional radiologic procedures or with Cone Beam Computed Tomography (CBCT) that allow a better 3D reconstruction of
the HCC nodule and its supplying vessels.
Aim: Our study aims to evaluate the efficacy of TACE performed
with CBCT or with traditional tecniques in the subpopulation of
patient in BCLC A ineligible to surgery.
Methods: Between January 2012 and June 2014 32 patients
were included in the study and 54 procedures were performed.
Patients in BCLC A treated with CBCT were 57% (12/21) and patient
treated with traditional technique were 27% (9/33).
Diagnosis of HCC is done considering EASL criteria and the radiologic response was evaluated at one month after treatment with the
RECIST score.
Results: The two groups were homogenous with no statistically
differences in their characteristics.
Patients in the CBCT group show a better response than patients
in traditional group (a 83% vs. 36%, p = 0.029). Patients in BCLC
B, instead, have similar response irrespective to the treatment
received (44% vs 54, CBCT and traditional group respectively, NA).
Conclusion: In BCLC A patients TACE performed with CBCT is
significantly more effective that the conventional procedure. This is
probably due to a more precise identification of the small nodule(s)
and blood supply. When the lesion is lager (group B) the advantage
of CBCT disappears.
T-33
CLINICALLY GUIDED ADJUSTMENT OF
SORAFENIB DAILY DOSE IS ADVISABLE FOR
CIRRHOTICS PATIENTS WITH HEPATOCELLULAR
CARCINOMA (HCC)
G. Gallusi, A. De Santis, C. Iegri, M. Lupo,
A. Mascolo, A.F. Attili
Department of Gastroenterology, La Sapienza
University of Rome, Rome, Italy
Introduction: Sorafenib is approved for treatment of advanced
HCC. Its efficacy/tolerability is often unpredictable. We looked for
prognostic factors of survival in a population of patients assuming
sorafenib for HCC.
Patients and methods: Ninety-three cirrhotic patients with HCC
were prospectively followed-up until last clinical control/death.
Commonest patients’ characteristics were: male sex (79,6%); viral
etiology (HCV/HBV) of cirrhosis (51,1%); ECOG PS 0 (68,9%) vs 1
(26,7%) and 2 (4,4%); BCLC C (60,4%) vs B (39,6%); presence of: portal vein thrombosis (57,8%), esophageal varices (57,1%), metastasis
(76,7%). Median age was 70 (45-88; 95% CI 6472); median ChildPugh score was 6 (range 5-9; 95% CI 5,79-6,20). Everybody started
therapy at 800 mg/day. If needed, posology was then reduced
according to patient’s tolerability and/or adverse events. Modifications of the posology/interruptions of treatment were registered. At
the end of observation, the dose assumed for ≥60% of the treatment
period was considered representative for each patient.
Results: In the entire population, median overall survival was
185 days (95% CI 161-256); median treatment duration 133 days
(95% CI 103-182). Skin toxicity (hand-foot skin reaction/rash) was
the most common adverse event (41,9%) and was associated to a
significantly higher median survival (396 vs 206 days, p = 0,02).
Fifty-four patients (58.1%) assumed sorafenib 800 mg/day and 39
patients (41.9%) 400 mg/day. The two groups did not differ for
the variables listed above. Probability of survival was significantly
higher with daily dose of 400 mg than 800 mg (p = 0,000488) (Figure 1). Accordingly, prevalence of skin toxicity was significantly
higher in patients treated with 400 mg/day than 800 mg/day. At
multivariate analysis, sorafenib dose and skin toxicity resulted
the only variables significantly associated to survival (respectively
p = 0,0009 and p = 0,0245).
Conclusions: When guided by clinical judgment of opportunity,
dose reduction should be considered feasible and maybe advisable,
especially in case of skin toxicity.
T-34
ANALYSIS OF THE ASSOCIATION OF PEDIATRIC
NAFLD WITH TWO SNPS OF GENES ENCODING
FOR FIBROBLAST GROWTH FACTORS 19 AND 21
RECEPTOR SYSTEM
Crudele, S. Ceccarelli, N. Panera, D. Gnani,
C. De Stefanis, A. Alisi, V. Nobili
Liver Research Unit, Bambino Gesù Children Hospital
and IRCCS, Rome, Italy
Introduction: Non-alcoholic fatty liver disease (NAFLD) is one
of most common liver diseases worldwide characterized by a
large spectrum of histological damages, including simple steatosis (NAFL), necro-inflammation and non-alcoholic steatohepatitis
(NASH), eventually associated with liver fibrosis. It is known that
NAFLD is a multifactorial disease caused by the interaction between
environmental factors and genetic background that trigger the
multiple mechanisms involved in disease pathogenesis. Fibroblast
growth factor(FGF)19 and FGF21 have been recently associated to
pediatric NAFLD suggesting a crucial role of their receptor system.
Aim: The aim of the present study was to investigate the
potential association of NAFLD with two single nucleotide polymorphisms (SNPs) of two genes encoding for proteins that compose the
FGF19/21 receptor system: the variant rs17618244 (Arg728Gln) of
Klotho-beta co-receptor (KLB) and the variant rs1966265 (Val10Ile)
of fibroblast growth factor receptor 4 (FGFR4).
Materials and Methods: To this purpose, we performed a casecontrol study including 100 Italian children without NAFLD and
150 with biopsy-proven NAFLD (74 with NAFL and 76 with NASH).
The SNPs were evaluated by a 5 -nuclease TaqMan assay.
Results: In the case-control study, the presence of NAFLD
increased with the number of G alleles for rs17618244 variant in
KLB (p < 0.01) and for rs1966265 variant in FGFR4 (p < 0.001). In
NAFLD children the severity of the disease (measured in terms
of the presence of NASH) was associated with several anthropometric and metabolic parameters of patients (body mass index,
cholesterol, transaminases and triglycerides), but not with a higher
presence of rs17618244 G allele. Interestingly, NAFLD children carrying the rs1966265 A allele had a trend for a higher prevalence of
NASH.
Conclusions: Our results suggest that the variant rs17618244 in
KLB and the variant rs1966265 in FGFR4 could be negatively associated with the presence of NAFLD even if FGFR4 SNP is associate
with the severity of disease.
T-35
T-36
RELATIONSHIP BETWEEN INFLAMMATORY
MEDIATORS AND FREQUENCY OF THE INNATE
IMMUNE CELLS IN PATIENTS WITH
NON-ALCOHOLIC FATTY LIVER DISEASE
THE USE OF A POCKET-SIZED ULTRASOUND
DEVICE (PUD) IMPROVES PHYSICAL
EXAMINATION: RESULTS OF AN IN- AND
OUTPATIENT STUDY
R. Maggio 1,3 , L. Antonucci 1 , C. Viscomi 1 ,
S. Costantini 2 , G. Castello 2 , C. Balsano 1,3
A. Colli 1 , D. Prati 2 , M. Fraquelli 3 , S. Segato 4 ,
P.P. Vescovi 5 , F. Colombo 6 , C. Balduini 7 ,
A. Baccarin 3 , S. Della Valle 8 , D. Conte 3 ,
G. Casazza 9 , for the Lombardy Ecoscopy Project
1
Centro Ricerche Oncologiche di Mercogliano,
Istituto Nazionale Per Lo Studio E La Cura Dei Tumori
“Fondazione Giovanni Pascale”, IRCCS, Mercogliano
(AV), Italy
3 Institute of Institute of molecular biology and
pathology (IBPM), National Research Council (CNR),
Rome, Italy
2
Nonalcoholic fatty liver disease (NAFLD) is the main liver diseaseworldwide. Simple steatosis remains a benign process, even if
it may progress to non alcoholicsteatohepatitis with necroinflammatory activity and inflammatory cell infiltration. Several studies
have demonstrated the involvement of proinflammatorymediators and innate immune cells in the NAFLD pathogenesis. We
investigated a possible relationship between circulating levels of
pro-inflammatory cytokines and percentage of immune cellular
innate components in NAFLD patients.
We enrolled 30 adult patients with a clinical diagnosis of
NAFLD and 10 healthy donors. The analysis ofimmune mediators
was performed on serum samples using multiplex immunoassays.Peripheral blood mononuclear cells were isolated from
patients and circulating ␥␦ T, NK, and NKT cells weremonitored by
flow cytometry. Correlation analysis was performed using Pearson
correlation test.
NAFLD patients showed elevated serum levels of chemoattractant proteins IL-8/CXCL8 and MIP1-␤/CCL4,compared to normal
subjects (p < 0.05). Levels of IL-8/CXCL8 correlated significantly
with MIP1-␤/CCL4levels, indicating a coordinated regulation of
their secretion (p = 0.001, r = 0.723). We observed a strong positivelinear correlation between IL-8/CXCL8 serum levels and ␥␦ T
cell-frequency (p = 0.001, r = 0.716). Moreover, IL-8levels were significantly associated with the percentage of NKT cells (p = 0.003,
r = 0.773), while nocorrelation was observed with the percentage of
NK cells. We revealed a markedly positive relationship between the
MIP1-␤/CCL4 serum levels and the frequency of ␥␦ T cells (p = 0.002,
r = 0.720). Thesame trend was obtained between MIP1-␤/CCL4
levels and the percentage of NKT cells (p = 0.005,r = 0.627).In conclusion, we demonstrated a direct correlation between the levels
of the pro-inflammatory cytokines IL-8/CXCL8 and MIP1-␤/CCL4,
and the cellular frequency of the innate immune system in NAFLD
patients. Thismight suggest that the levelsof the pro-inflammatory
cytokines investigated could represent a “substratum” for the
immunological events occurring during the NAFLD progression.
e35
1 Dipartimento Medicina Interna, Ospedale A
Manzoni, Lecco, Italy
2 Dipartimento Medicina Trasfusionale e Ematologia,
Ospedale A Manzoni, Lecco, Italy
3 Unità di Gastroenterologia ed Endoscopia, IRCCS
Fondazione Ca Granda Ospedale Maggiore
Policlinico, Università degli Studi di Milano, Italy
4 Unità di Gastroenterologia ed Endoscopia, Azienda
Ospedaliero Universitaria Macchi, Varese, Italy
5 Divisione di Medicina Interna, Azienda Ospedaliera
“Carlo Poma” Mantova Italy
6 Prima Divisione di Medicina Interna, A.O.Niguarda
Milan, Italy
7 Terza Divisione di Medicina Interna, Fondazione
IRCCS Policlinico San Matteo-Università degli Studi,
Pavia, Italy
8 Dipartimento di Medicina Interna, Ospedale A
Manzoni Lecco, Italy
9 Dipartimento di Scienze Biomediche e Cliniche,
Ospedale L. Sacco, Universita degli Studi di Milano,
Milano, Italy
Background/aim The performance of PUD is comparable with
that of standard ultrasonography, whereas the accuracy of physical
examination (PE) is often poor requiring further tests to assess diagnostic hypotheses. Adding PUD to PE could lead to an incremental
benefit. Present cohort-impact study aimedat assessing whether
the use of PUD in the context of PE could reduce the request rate
of additional tests when used by physicians in different clinical
settings
Methods The study involved medical wards(#4), hepatological
outpatient clinic(#1) and 90 GPs operating in the same geographical area. After a short training course 135 physicians used PUD
in addition to PE in 1962 consecutive patients to investigate
ten well-defined clinical hypotheses (ascites, biliary-duct dilation/gallstones, splenomegaly, pleural/pericardial effusion, urinary
retention/stones, abdominal mass/aortic aneurysm. PUD-related
findings were recorded. and the decision as to whether to request
further tests was left to the physician clinical judgement. The main
outcome was to define the proportion of cases undergoing additional tests after PUD. An accurate report of the frequency of the
clinical questions in the different settings was also planning, using
logistic regression analysis to assess the determinant(s) of the primary outcome.
Results Of the 1962 included patients 726(37%) were inpatients,
510(26%) hepatology outpatients and 726(37%) recruited from GPs.
Gallstone (37%), ascites (17%), pleural effusion (13%), urinary stones
(13%) and urinary retention(12%) accounted for > 90% of the clinical
questions, confirmed by PUD in 66% of cases. The overall frequency
of further tests after PUD was 37%; and logistic was associated with
both the clinical questions and settings(p < 0.01). The rate of agreement between PUD findings and additional tests was 89% (Sens
91%,Spec 83%,LR + 5.4;LR-0.11).
Conclusions After a short training, PUD examination can be
used in addition to PE in in- and outpatients to improve the answer
e36
to ten common clinical questions, thus reducing the need further
testing.
T-37
ADHERENCE TO EASL-EORTC CLINICAL
GUIDELINES FOR THE MANAGEMENT OF
HEPATOCELLULAR CARCINOMA IN FIELD
PRACTICE: RESULTS FROM THE ITALICA
DATABASE
R. Sacco, V. Mismas, L. Giacomelli, S. Marceglia,
A. Romano, M. Bertini, M. Bertoni, G. Federici,
G. Parisi, S. Metrangolo, E. Tumino, G. Bresci
Department of Gastroenterology, Pisa University
Hospital, Pisa, Italy
Introduction: Data on adherence to joint guidelines for the
management of hepatocellular carcinoma (HCC) published in 2012
by the European Association for the Study of the Liver (EASL) and
the European Organization for Research and Treatment of Cancer
(EORTC) are lacking.
Aim: We retrospectively evaluated the adherence to EASLEORTC guidelines in field-practice, using data from HCC patients
registered in the Nation-wide Italian database ITA.LI.CA. and diagnosed from 2012.
Methods: The ITA.LI.CA. database contains data of 5428 HCC
patients treated at 18 Italian Centers. Patients were stratified
according to Child-Pugh (CP) and and the Barcelona Clinic Liver
Cancer (BCLC) classifications. We investigated the adherence to
surveillance, diagnosis, and first-line treatment recommendations.
Results: In ITALICA, 600 patients were diagnosed of HCC since
2012 (466 males; mean ± SD age 67.4 ± 10.9 years; 277(46.2%)
CP-A and 163(27.2%) CP-B; 44(8%) BCLC-0, 193(35.1%) BCLC-A,
93(16.9%) BCLC-B, 172(31.3%) BCLC-C, 48(8.7%) BCLC-D). Overall, 317(55.2%) were diagnosed during a surveillance program. Of
them, 231(57.9%) were cirrhotic (median surveillance duration: 6
months). Four-hundred-ninety-six (85.3%, 449 cirrhotic) patients
were diagnosed applying a radiological, 80(13.7%) a histological,
and 6(1%) a cytological criterion. Five (9.7%) patients in BCLC stage 0
with CP A, and single nodules underwent tumour resection; 3(1.4%)
patients in BCLC-A received liver transplantation, and 83(43.1%)
received radiofrequency ablation or Percutaneous Ethanol Injection. Intermediate HCC-stage patients (BCLC-B) receiving TACE
were 45(47.9%), and advanced-stage patients (BCLC-C) receiving
sorafenib were 38(21.9%). Palliative care was provided to terminal
stage patients (BCLC-D) in 31(64.3%) cases.
Conclusions: The overall adherence in a “real-world” practice to
EASL-EORTC guidelines was low, particularly in patients with early
stage HCC. Difficulties inpatients staging and the high prevalence of
older patients with relevant co-morbidities may partially explain
these findings. Strategies to help improve adherence to international guidelines for HCC in field-practice and new scoring criteria
are required.
T-38
CLINICAL PATTERNS OF HEPATOCELLULAR
CARCINOMA (HCC) IN NON ALCOHOLIC FATTY
LIVER DISEASE (NAFLD): A MULTICENTER
CASE-CONTROL STUDY
F. Piscaglia 1 , G. Svegliati Baroni 2 , A. Barchetti 3 ,
A. Pecorelli 1 , S. Marinelli 1 , C. Tiribelli 3 ,
S. Bellentani 3 e gruppo di studio italiano HCC
NAFLD composto da: L. Bolondi, M. Zoli,
D. Malagotti, G. Brandi, E. Bugianesi, E. Vanni,
L. Mezzabotta, G. Cabibbo, S. Petta, A. Fracanzani,
S. Fargion, F. Marra, B. Fani, R. Sacco, F. Morisco,
N. Caporaso, M. Guarino, M. Colombo,
R. D’Ambrosio, L.S. Crocè, R. Patti, E. Giannini,
A. Lonardo, E. Baldelli, L. Miele, A. Grieco,
F. Farinati, C. Pozzan, M. Borzio, E. Dionigi,
G. Soardo, P. Roselli, F. Ciccarese, F. Virdone,
A. Affronti, F.G. Foschi, F. Borzio, F. Trevisani
1
Unità di Medicina Interna, Dipartimento di Scienze
Mediche e Chirurgiche, Policlinico S.
Orsola-Malpighi, Università of Bologna
2 Unità di Gastroenterologia, Università Politecnica
delle Marche, Ancona
3 Centro Studi Fegato (CSF), Liver Research
Center–Parco della Scienza - Basovizza Campus,
Trieste
Background The prevalence of non-alcoholic fatty liver disease
(NAFLD) is growing and the disease can progress to hepatocellular
carcinoma (HCC). Only scant clinical information on HCC in NAFLD
is available.
The aim of the study was to assess the clinical features of patients
with NAFLD-related HCC (NAFLD-HCC), and to compare them to
those having HCV-related HCC.
Methods: The study is a multicenter observational case-control
study. 756 patients with either NAFLD (145) or HCV-related chronic
liver disease (611) have been enrolled in Secondary Care Italian
Centers. Survival was modeled according to clinical parameters,
lead time bias and propensity analysis.
Results: Patients with NAFLD- HCC were: less often male than
HCV-HCC (61% vs 79%), had a higher BMI (29.1 vs 27.6), similar
Child-Pugh score (5.6 vs 5.8), larger tumor volume (largest nodule
4.1 vs 3.3 cm), more often infiltrative pattern (15.4% vs 4.0%) and
detection outside specific surveillance (52.4% vs 36.7%). Survival
was significantly shorter (p = 0.017) in NAFLD-HCC: 25.5 months
(95% CI 21.9-29.1) than in HCV-HCC patients 33.7 months (95% CI
31.9-35.4). This difference remained significant even after adjustment for lead time bias in surveilled patients. Cirrhosis was present
in only about 50% of NAFLD-HCC patients. To understand the intrinsic aggressiveness of HCC and to eliminate possible confounders, a
propensity score analysis was carried out considering age, gender,
Child-Pugh score, largest tumor size, leaving 64 patients in each
group. The difference in mean surviva between the two groups was
no longer statistically significant (30.2 months in NAFLD-HCC and
36.9 HCV- HCC, p = 0.330).
Conclusions NAFLD-HCC rises in the absence of cirrhosis in
about 50% of the cases, and it is detected at a later tumor stage than
in HCV-infected patients. Future research should identify patients
with NAFLD who require surveillance in order to offer them the
most timely and effective treatment.
T-39
FAS-LIGAND INVOLVEMENT IN END STAGE LIVER
DISEASE
E. Trombetta 1 , P. Cetrangolo 1 , A. Cattaneo 1 ,
G. Rossi 2 , D. Prati 3 , L. Porretti 1 , F. Colombo 1,∗
1 Flow Cytometry Service, Fondazione IRCCS Ca’
Granda Ospedale Maggiore Policlinico di Milano,
Milan, Italy
2 Liver Transplant Unit, Fondazione IRCCS Ca’ Granda
Ospedale Maggiore Policlinico di Milano, Milan, Italy
3 Department of Transfusion Medicine and
Haematology, Ospedale “A. Manzoni”, Lecco, Italy
Introduction: FasL has been involved in lymphocyte function
and disease development. Recently, it has been demonstrated that
CD8 + /FasL + cells can be eliminated by tumor cells as escape mechanism.
Aim: To evaluate if CD8 + /FasL + lymphocytes are also involved
in end stage liver disease.
Materials and Methods: 6 blood donors and 4 cirrhotic patients
were enrolled. FasL expression on CD8 + cells of donors and patients
peripheral blood (PB) was evaluated by flow cytometry before
and after 24 h of activation with PMA and ionomycin. Moreover,
we evaluated FasL expression on CD8 + cells infiltrating patients’
liver parenchyma in bioptic specimens. To mimic lymphocyte-liver
parenchyma interaction, we implemented co-cultures with lymphocyte and HCC cell lines. Donor PBMC, either before or after 4 h of
activation, were co-cultured with 2 FasL + HCC cell lines (ratio 1:5,
18 h) or alone in the same conditions. Cell mortality was evaluated
by means of 7AAD and Zombie Aqua staining.
Results: The percentage of PB CD8 + cells was equal between
patients and controls, while FasL expression on CD8 + cells was
lower in patients (34.2 ± 4.5 vs. 51.4 ± 5.9, p < 0.01). Similar data
were obtained after 24 h of activation, although this process did not
increase FasL expression on CD8 + cells. Interestingly, liver infiltrating CD8 + cells expressed less FasL in comparison to the circulating
counterpart (17.5 ± 9.5 vs. 34.2 ± 4.5, p = 0.032), although no differences in CD8 + percentage were observed between PB and liver
parenchyma (18.4 ± 13.2 vs. 33.4 ± 4.8, p > 0.05). Finally, an equal
CD8 + /FASL + cell mortality was observed when resting CD8 + cells
were co-cultured with HCC cell lines (13.3 ± 3.0, 12.7 ± 4.1 vs.
10.3 ± 7.9), while co-cultures with activated CD8 + cells produced a
higher CD8 + /FASL + cell death (47.1 ± 12.5, 43.9 ± 5.2 vs. 25.1 ± 8.3,
p < 0.05).
Conclusions: Our data suggest that diseased liver cells could
be responsible for CD8 + /FasL + depletion by an escape mechanism,
both within the liver parenchyma and in PB, favouring the progression of the disease.
e37
T-40
NEUTROPHIL FUNCTION IN PATIENTS WITH
CHRONIC HEPATITIS C (CHC) UNDERGOING
TRIPLE ANTIVIRAL THERAPY (TT) WITH FIRST
AND SECOND GENERATION PROTEASE
INHIBITORS (PI)
M. Gambato 1,2 , N. Caro-Pérez 1 , N. Cañete 3 ,
Z. Mariño 1 , S. Lens 1 , JM. Sánchez-Tapias 1 ,
J. Carrion 1 , S. Pérez-del-Pulgar 1 , M. Juan 4 ,
M. Londoño 1 , X. Forns 1
1 Liver Unit, Hospital Clínic, IDIBAPS, Ciberehd,
Barcelona, Spain
Surgery, Oncology and Gastroenterology,
PaduaUniversityHospital, Padua, Italy
3 Digestive disease Unit, Hospital del Mar, Barcelona,
Spain
4 Immunology Unit, Hospital Clinic Barcelona, Spain
Introduction Cirrhotic patients undergoing TT with pegIFN/RBV and a first generation PI have higher risk of bacterial
infections as compared to those receiving IFN/RBV (Londoño et al. J
Hepatol 2014). PIs might inhibit neutrophil proteases and Toll-like
receptor (TLR) pathways.
Aim We aimed at evaluating innate immune recognition and
neutrophil activity in CHC patients undergoing TT.
Materials and Methods: CHC patients undergoing TT after a
lead-in phase were prospectively studied. Blood samples were
obtained at baseline, week 4 and week 8. Phagocytic and oxidative burst activities were quantified by flow cytometry. Neutrophils
were isolated from peripheral blood and stimulated with flagellin
(FLA) to determine cytokine secretion (IL-1b, IL-6, IL-10, IL-12).
Quantification was performed by Luminex®.
Results: 18 patients have been enrolled so far (4 F2, 3 F3 and 11
F4). PIs used were telaprevir (n = 9), boceprevir (n = 1) or simeprevir
(n = 8). The number of phagocytized bacteria per cell, determined
by the mean fluorescence intensity (MFI), significantly decreased
at weeks 4 and 8 as compared to baseline (p = 0.02 and p = 0.035,
respectively). However, there were no significant differences in MFI
between weeks 4 and 8. Oxidative burst capacity did not change
throughout the study period. Cytokine levels measured after neutrophil stimulation with FLA were increased at week 4 and highly
decreased at week 8 of therapy, but the difference was not statistically significant (p = 0.3 for all cytokines).
Conclusions: Antiviral therapy with peg-IFN/RBV is associated
with a significant decrease in neutrophil phagocytic capacity, but
the addition of a PI does not seem to further reduce this function.
Interestingly, after 4 weeks of PI therapy, there was a decrease in
cytokine secretion which might suggest an inhibition of proteases
involved in TLR signaling pathways. The combination of both effects
could explain the development of infections previously described
in cirrhotic patients undergoing TT.
e38
T-41
T-42
SMALL FIBERS PERIPHERAL NEUROPATHY IN
WILSON’S DISEASE: AN IN VIVO
DOCUMENTATION BY CORNEAL CONFOCAL
MICROSCOPY
HEPATIC VENOUS PRESSURE GRADIENT IN THE
PREOPERATIVE ASSESSMENT OF PATIENTS WITH
RESECTABLE HEPATOCELLULAR CARCINOMA
G.C.
O.
D.
M. Berton 2 , A. Leonardi 2 , I.A. Fregona 2 ,
E. Midena 2
A. Cucchetti 1 , M. Cescon 1 , F. Piscaglia 1 ,
R. Golfieri 2 , M. Renzulli 2 , C. Mosconi 2 ,
A. Cappelli 2 , F. Mazzotti 1 , F. Neri 1 , G. Ercolani 1 ,
A.D. Pinna 1
1
1
Sturniolo 1 ,
Bartolo 1 ,
Lazzarini 2 ,
Department of Gastroenterology, University of
Padua, Padua, Italy
2 Department of Ophthalmology, University of
Padua, Padua, Italy
Wilson disease (WD) is an inherited autosomal recessive disorder of copper metabolism. Corneal confocal microscopy (CCM) is
a fast technique to analyze the human cornea in vivo. We aimed
to investigate central corneal changes and to assess the parameters of corneal subbasal nerve plexus (CSNP) in patients affected
by WD, using CCM. 24 patients affected by WD and 24 healthy
control subjects were enrolled in this cross-sectional comparative
study. One eye of each subject was examined to quantify different
corneal parameters, by CCM. WD induced significant alterations
in both corneal subbasal nerve plexus, and corneal epithelium. All
the parameters of the subbasal nerve plexus were altered in WD,
documenting, for the first time, a (corneal) peripheral nerve damage in WD. The decrease of major CSNP parameters confirms the
damage (and death) of a significant number of small nerve fibers,
whereas the increase of fiber tortuosity is a sign of tentative nerve
regeneration. Mean epithelial cell diameter (P < 0.0001) and mean
epithelial cell density (P < 0.0001) resulted significantly higher and
lower compared to controls, respectively. No significant difference
in corneal stroma and endothelium were observed; no significant
differences were documented among different treatment groups.
The comparison between predominantly neurological and predominantly hepatic presentation didn’t show any statistical differences.
The only difference in subgroup was found between patients with
previous of KF ring e without KF ring. CCM showed significant
corneal changes in subbasal nerve plexus, and secondary corneal
epithelium changes in WD. CCM clearly documented for the first
time, a (corneal) peripheral nerve damage in WD. Corneal damage
was more severe in pts with previous KF ring. CCM may contribute
to the diagnosis and monitoring of the peripheral nervous system
involvement in WD. Further larger studies needed to confirm these
findings.
S.Orsola-Malpighi Hospital, Alma Mater Studiorum University of Bologna, Bologna, Italy
2 Department of Diagnostic Medicine and
Prevention, S.Orsola-Malpighi Hospital, Alma Mater
Studiorum - University of Bologna, Bologna, Italy
Background: According to guidelines, hepatic resection (HR) for
hepatocellular carcinoma (HCC) should be offered to patients without significant portal hypertension (PH), namely, with a hepatic
venous pressure gradient (HVPG) < 10 mmHg. In real life, HCC
patients with clinical signs of PH are often referred for surgical
evaluation and eventual hepatectomy.
Methods: Data from 70 consecutive, prospectively enrolled,
HCC patients submitted to HR were collected and analyzed to define
the role of HVPG in the prediction of post-operative liver failure
(PHLF) grade B/C, as defined by the International Study Group of
Liver Surgery.
Results: Post-operative and 90-days mortalities were null. The
median HVPG value was 9 mmHg (range: 4-18), median MELD
was 8 (range 6-14); 34 patients had HVPG≥10 mmHg (48.6%).
Forty-nine patients experienced an uneventful (Grade A) postoperative course; of them, 17 had HVPG≥10 mmHg (24.2% of
70 patients). Grade B complications occurred in 20 patients (3
with HVPG < 10 mmHg and 17 with HVPG≥10 mmHg; P < 0.001);
only one grade C complication occurred in a patient having
HVPG < 10 mmHg, subsequently successfully submitted to liver
transplantation. Receiver Operating Characteristics curve for PHLF
B/C showed for Model for End-stage Liver Disease (MELD) an area
under the curve (AUC) of 0.727 (95%C.I.: 0.598–0.857) and for HVPG
of 0.792 (95%C.I.: 0.677–0.907; P = 0.475).
Conclusions: Hepatic venous pressure gradient can be used
before surgery to stratify the risk of PHLF but does not add significant further prognostic ability with respect to MELD score. The
proposed cut-off of 10 mmHg is indeed associated with worse prognosis but excludes about one quarter of patients who will benefit
from surgery without short-mid term post-operative sequelae.
T-43
T-44
HEPATITIS DELTA PREVALENCE AND CLINICAL
EXPRESSION IN AN IMMIGRANT POPULATION IN
ITALY
PNPLA3 VARIANT IS AN INDEPENDENT
PREDICTOR OF SEVERE STEATOSIS IN PATIENTS
WITH CRONIC HEPATITIS C AND HIV INFECTION
G. Colucci 1 , M. Colombo 1 , G. Lunghi 2 , P. Bono 2 ,
S. Fadelli 3 , M. Adamoli 3 , D. Aurelio 3 ,
M. Abbruzzese M 3 , R. Romeo 1
C. Sagnelli 1 , M. Merli 2 , C. Uberti-Foppa 2 ,
A. Gradone 3 , H. Hasson 2 , G. Cirillo 3 ,
S. Salpietro 2 , C. Minichini 4 ,
E. Miraglia Del Giudice 3 , A. Lazzarin 2 ,
E. Sagnelli 4 , N. Coppola 4
Fondazione Ca’ Granda, Policlinico, Milan, Italy
2 Laboratory medicine Fondazione Ca’ Granda,
Policlinico, Milan, Italy
3 Poliambulatorio Opera San Francesco per i Poveri
(OSF), Milan, Italy
Background and aims: Hepatitis delta virus (HDV),a defective
agent coinfecting 5% of chronic hepatitis B virus (HBV) carriers, has
recently regained the interest of the scientific community because
of its increasing prevalence and morbidity rates, particularly in
developing countries. In this respect, we started a screening program at one of the largest NGO clinic in Italy, Opera San Francesco
(OSF) in Milan where patients are mainly illegal immigrants or
refugees lacking health insurance coverage and often proper housing accommodation.
Methods: Using commercially available tests (Diapro Srl, Sesto
S. Giovanni, Italy) HBV and HDV infections were sought in consecutive, unselected individuals, independently on their health status.
Following HBsAg and IgG and IgM anti-HDV detection, positive
patients underwent HBVDNA and HDV RNA testing whereas the
screening tests were offered to their close relatives.
Results: Starting September 2014, 119 migrants originating
from North Africa (10%), Eastern Europe (48%), Central and South
America (33%) and Asia (9%) have been screened. 23 patients
resulted HBsAg positive of whom two patients with compensated
cirrhosis tested also IgG and IgM anti-HDV positive but HDV RNA
undetectable (< 30 copies/ml) and are currently treated with Entecavir, 300 mg/die.
Conclusion: This interim analysis suggests a high HBsAg carrier state prevalence (19.3%) and HBV/HDv co infection rate of 8.6%
among migrants slightly lower than that recently reported, particularly in east Europe, but probably better reflecting the situation of
the general population in these countries as we did not apply any
patients selection. By the time this screening project is concluded it
will be the largest ever carried out and the most informative on the
actual HBsAg and HDV prevalence in a mixed immigrant population
from high endemic countries.
e39
1 Department Experimental Medicine and Surgery “F.
Magrassi e A. Lanzara”, Second University of Naples,
Naples, Italy
2 Department of Infectious Diseases, Vita-Salute
University,San Raffaele Scientific Institute, Milan,
Italy
3 Department of Pediatrics, Second University of
4 Department Mental Health and Public Medicine,
Background A correlation between the p.148 polymorphism
of the patatin-like phospholipase domain-containing protein 3
(PNPLA3) and the degree of liver steatosis in patients with chronic
hepatitis C (CHC) was identified.
Aim: to analyze the impact of PNPLA3 variants on liver histology
of CHC patients with HIV infection.
Methods: 168 consecutive patients with HIV infection and
biopsy proven CHC, naïve for HCV-treatment, HBsAg-negative and
with no alcohol abuse were enrolled. Patients aged 40.7 ± 6.0, 72.6%
were males, 42% with HCV-genotype 3 and 38.9% with genotype
1, 81.5% in HAART; the nadir of CD4 + was 290.0 ± 197.4 cell/␮L.
Liver fibrosis and necroinflammation were analyzed by the Ishak’s
scoring system and steatosis as follow: score 1 = fatty deposition
in 1-10% of hepatocytes, score 2 in 11-30%; score 3 in 31-60% and
score 4 in > 60%.
Results: No difference was observed in baseline demographic,
laboratory characteristics and degree of necroinflammation and
fibrosis between patients with different PNPLA3 variants. The 79
subjects with PNPLA3 I/M-M/M variants showed a severe steatosis (degree 3-4) more frequently than the 89 with PNPLA3 I/I (43%
vs 24.7%, p = 0.001). Compared with the 112 patients with steatosis score 1-2, the 56 with score 3-4 had higher BMI (23.91 ± 2.99
vs 22.88 ± 2.82, p = 0.03), ALT serum level (106.84 ± 63.11 vs.
70.53 ± 60.16, p = 0.0002), higher prevalence of cases with HCVgenotype 3 (55.6% vs. 35.2%, p = 0.01) and of PNPLA3 I/M-M/M
(60.7% vs 39.3%, p 0.01).
A multivariate logistic regression analysis including PNAPL3
variants (I/M or M/M vs I/I), HCV genotype (3 vs. other genotypes),
BMI and other potential confounding factors (age, sex) showed
a high BMI (0.0165), HCV-genotype 3 (p = 0.0075) and PNPLA3
p.I148 I/M-M/M polymorphism (p = 0.0088) independently associated with liver steatosis score 3-4.
Conclusions: This study is the first demonstration that the
PNPLA3 p.148 I/M-M/M variants is an independent predictor of
severe liver steatosis in CHC patients with HIV infection
e40
T-45
T-46
PROACTIVE DOSE ADJUSTMENTS ARE
NECESSARY IN MANY ADV-EXPERIENCED
PATIENTS TREATED WITH TDF MONOTHERAPY
FOR 5 YEARS: A PROSPECTIVE COHORT STUDY IN
320 PATIENTS
TOTAL HEPATITIS B CORE ANTIGEN ANTIBODY, A
QUANTITATIVE NON-INVASIVE MARKER OF
HEPATITIS B VIRUS INDUCED LIVER DISEASE
P. Lampertico 1 , R. Soffredini 1 , M. Borghi 1 ,
M. Viganò 2 , F. Facchetti 1 , E. Galmozzi 1 ,
F. Invernizzi 1 , M. Colombo 1
1
Introduction: Tenofovir (TDF) is a popular anti-HBV strategy
for NUC-experienced patients, but its safety profile in field practice
patients previously exposed long-term to Adefovir (ADV) is under
discussion because of recent reports of Fanconi syndrome.
Materials and Methods Results: 320 NUC-experienced CHB
patients received Tenofovir (TDF) for 69 months (range 1-106) as
a switch from ADV + LAM (86%) or as a rescue therapy for LAM,
ADV or ETV (age 59, 85% HBeAg-negative, 62% cirrhotics, 88% normal ALT, 74% undetectable HBV-DNA). Safety analysis focused on
glomerular (eGFR) and tubular renal function. Baseline was the start
of TDF. During 5 years of TDF, 89 patients (28%) had to reduce TDF
dose after 31 months (2-75) (57 for eGFR decline, 30 for phosphate
decline, 2 for both events). 11 additional patients (3.4%), who had to
stop TDF because of low blood phosphate levels, were successfully
rescued by switching to ETV. 6 (2%) additional patients withdrew
for gastrointestinal side effects. Overall, 106 patients (33%) either
required a dose reduction or withdrew from TDF for side effects,
with a 5-year estimated probability of 40%. None of the TDF patients
developed acute renal failure or experienced a virological rebound
after dose adjustment. Virological response progressively increased
to 100% at year 5. 15 patients (5%) cleared HBsAg, 26 additional
patients had qHBsAg < 10 and 25 reached qHBsAg between 10 and
100 IU/ml (Overall, 22% with qHBsAg < 100 IU/ml). HCC attack rates
were 1.3%/year in compensated cirrhotics and 0.2%/year in non cirrhotics. No cases of clinical decompensation were recorded. Overall,
7 patients (2%) were transplanted (all for HCC) and 16 (5%) died (7
for HCC, 7 for non liver causes and 2 for unknown reasons).
Conclusions: To prevent glomerular and tubular damage,
proactive dose adjustments of TDF are necessary in a large proportion of ADV-experienced, TDF treated CHB patients.
F. Moriconi 2 , Q. Yuan 1 , L-W. Song 1 ,
D. Cavallone 2 , B. Cherubini 2 , P. Colombatto 2 ,
F. Oliveri 2 , B. Coco 2 , G. Ricco 2 , F. Bonino 3 ,
J.W-K. Shih 3 , N-S. Xia 1 , M.R. Brunetto 2
1
State Key Laboratory of Molecular Vaccinology and
Molecular Diagnostics, National Institute of
Diagnostics and Vaccine Development in Infectious
Diseases, School of Public Health, Xiamen University,
Xiamen, China
2 Laboratory of Molecular Genetics and Pathology of
Hepatitis Viruses, Hepatology Unit, Reference Center
of the Tuscany Region for Chronic Liver Disease and
Cancer, University Hospital of Pisa, Italy
3 Digestive and Liver Disease, General Medicine II
Unit, University Hospital of Pisa, Italy
Background and Aim Non invasive immunologic markers
of virus-induced-liver-disease are an unmet need. We compared the performances of quantitative total and IgM-anti-HBc in
well-characterized chronic-HBsAg-carriers. Methods. Sera (212)
were obtained from 111 HBsAg-carriers followed-up for 40
months (18-216 months) during different phases of chronic-HBVgenotype-D-infection; 10 HBeAg-positive, 25 inactive-carriers
(HBV-DNA≤2000IU/ml, ALT < 30 U/L), 66 HBeAg-negative/antiHBe-positive-CHB-patients and 10 with HDV-super-infection. In
35 patients treated with Peg-IFN (180 ␮g/w for 12 m) sera were
obtained at baseline, end-of-therapy and week-24 off-therapy
and in 22 treated with nucleos(t)ide-analogues (for 60 months
mean, 42-134 months range) at baseline and end-of-follow-up.
IgM- and total-anti-HBc were measured by Architect (Abbott, USA)
and double-antigen-sandwich-immune (Wantai, China) assays
respectively. Results. Total-anti-HBc was positive in all sera
with lower levels (p < 0.001) in HBsAg-carriers without CHB
(immune-tolerant, inactive and HDV-superinfected, mean 3.26,
range 2.26-4.49 Log10 IU/ml) versus untreated-CHB (mean 4.68,
range 2.76-5.54 Log10 IU/ml). Thirty of 212 (14.2%) sera were
IgM-anti-HBc-positive using acute-hepatitis-cut-off (1-S/CO value)
and 102 (48.1%) with chronic-hepatitis-cut-off (0.130-S/CO). Totaland anti-HBc-IgM correlated (r2 = 0.4173). Total-anti-HBc declined
significantly in CHB-patients with sustained virological response
(p < 0.001) treated with antivirals: groups a) inactive-carriers, b)
baseline-untreated-HBeAg-negative-CHB, c) EOF-HBeAg-negativeCHB with SVR after Peg-IFN, d) EOF in NUC-treated-patients]; the
lowest levels were found in SVR who cleared HBsAg subsequently.
The distribution of total-anti-HBc and the differences between
groups are clinically significant with values in responders to antiviral therapy (c and d) comparable to inactive carriers. IgM-anti-HBc
values were in most of sera below the analytical-specificity cut-off
of the assay even using the lower CHB cut-off of 0.130 S/CO. During
spontaneous and therapy-induced CHB remissions and reactivations both total- and IgM-anti-HBc correlated with ALT (p < 0.001,
r = 0.351 and p = 0.008, r = 0.185 respectively). Conclusions. Totalanti-HBc qualifies as a useful marker of HBV-induced-liver-disease
that might help to discriminate major phases of chronic HBV infection and to predict sustained response to antivirals.
T-47
BOCEPREVIR OR TELAPREVIR PLUS
PEGINTERFERON/RIBAVIRIN IN HCV CHRONIC
INFECTION: THE REAL-LIFE EXPERIENCE OF THE
ITALIAN ASSOCIATION OF HOSPITAL
HEPATOLOGISTS (CLEO)
e41
the mean follow-up was 103.8 ± 86 months. Forty-nine patients
(53.3%) had associated IBD (38 ulcerative colitis, 10 Crohn’s disease,
1 indeterminate colitis). Clinical activity and endoscopic severity
were scored according to the CD activity index and Mayo subscore
for endoscopy.
Results: Table 1 shows the major events occurred during the
follow-up:
CLEO DAAs Study Group, CLEO, Rome, Italy
Introduction: Boceprevir/Telaprevir (DAAs), approved for
reimbursement in Italy in December 2012, were used from January
2013. Since then the group of the Association of Hospital Hepatologists (CLEO DAAs Study Group) has been deeply involved in using
DAAs. In September 2013, the Governing Board of the Association
decided to collect data from Hospital Centers, where there were
active members belonging to CLEO. For this reason, this study can
be qualified as retrospective/prospective.
Aim: to check safety and efficacy of this type of treatment in the
real-world setting.
Patients and Methods: A database was prepared and used by
all Centres for the data collection and updated continuously. Last
update: November 26, 2014. All patients consecutively treated
were included; data were analyzed according to the intention-totreat principle. HCV-RNA was analyzed using: COBAS TaqMan 2.0
(Roche) with LLQ 25IU/mL.
Results: 40 Centres enrolled 737 patients: male 64%; median
age 58 (range 18-78), of whom 20.2% over 65; mean BMI 25.6
(range 16-39); Genotype 1b (78.4%); fibrosis F3/4 (70%). DAAs used:
Telaprevir (67%); PEGIFN-2a (71%); patients naïve (28%), relapsers
(32%), non-responders (40%). Therapy was stopped in 13.4% cases
because of side-effects (anaemia 37%, rash 26%) or for virological
failure (16.2%). Since the study is ongoing, we have 563 patients
who completed the follow-up. The RVR was achieved in 68% cases,
EOT in 64%, while the SVR was achieved in 68% in F0-F1; 57% in
F2-F3 and 37% in F4. In cirrhotic aging > 65 the SVR was 34%. There
were no fatalities.
Conclusions: DAAs, in everyday practice, are safe but with moderate efficacy. These data confirm the limited success of DAAs in
certain groups of patients such as those widely represented in our
series: advanced fibrosis/cirrhosis, non-responders to PEGIFN/RIBA
and the over 65s. As for the SVR, the grade of fibrosis makes the
difference.
T-48
EFFECTIVENESS OF ADALIBUMAB FOR PATIENTS
WITH PRIMARY SCLEROSING CHOLANGITIS
ASSOCIATED WITH INFLAMMATORY BOWEL
DISEASE
I. Franceschet, N. Cazzagon, A. Floreani
Department of Surgery, Oncology and
Background: Primary sclerosing cholangitis (PSC) is a chronic
biliary disease with a marked comorbidity with extra-hepatic conditions, mainly inflammatory bowel disease (IBD). The medical
treatment for PSC is still disappointing, whereas immunomodulators and biologics have been able to demonstrate efficacy in IBD.
Aim: To analyse: 1) the natural history of patients with
PSC ± associated with IBD; 2) the long-term efficacy of biologics
in patients with PSC and concomitant IBD.
Methods: 92 consecutive PSC patients (seen from 1987 to 2014)
were included in the study: 50 (54.3%) were males, and 42 (45.7%)
females. The mean age at diagnosis was 32.0 ± 14.3 years, and
Events
PSC + IBD (N.49)
PSC-IBD (N.43)
p
OLTx
CCA
HCC
Gallbladder cancer
Colo-rectal cancer
Death
7 (14.3%)
0
2 (4.1%)
1 (2.0%)
4 (8.2%)
5 (10.2%)
7 (16.3%)
2 (4.6%)
0
0
0
5 (11.6%)
0.79
0.13
0.18
0.35
0.05
0.83
Three patients with IBD experienced, as second-line treatment,
Adalimumab (ADA), an anti-TNFa monoclonal antibody, previous
written consensus. Patients were assessed before starting treatment, at month 6 and 12. ADA induction was 160 mg at week 0,
and then 80 mg at week 2, while ADA maintenance treatment was
40 mg every 2 weeks. After 6 and 12 months of ADA, a sustained
clinical remission of IBD was obtained; a reduction in ALT, GGT,
alkaline phosphatase and Mayo PSC score was obtained.
Conclusions: This is the first study evaluating the efficacy of biologic agents in PSC. Promising results come from ADA for PSC + IBD
during a 12 months follow-up. Furthers studies are warranted to
investigate the long-term tolerability and efficacy in such patients.
T-49
POSITIVE CORRELATION OF HIF2␣ AND
SERPINB3 IN HUMAN HEPATOCELLULAR
CARCINOMA: SELECTIVITY AND PROGNOSTIC
IMPLICATIONS
E. Morello 1 , C. Turato 2 , S. Cannito 1 ,
M. Ruvoletto 2 , S. Quarta 2 , C. Paternostro 1 ,
E. Novo 1 , R. Autelli 1 , S. Fasolato 2 , I. Tusa 3 ,
E. Rovida 3 , S. Colombatto 4 , A. Smedile 5 ,
A. Vitale 6 , G. Zanus 6 , U. Cillo 6 , M. Parola 1 ,
P. Pontisso 2
1 Department Clinical and Biological Sciences, Unit of
Experimental Medicine, University of Turin, Turin,
Italy
2 Department Medicine, University of Padua, Padua,
Italy
3 Department Biomedical, Experimental and Clinical
Sciences, University of Florence, Florence, Italy
4 Department Oncology and
5 Department Medical Sciences, University of Torino,
Italy
6 Department Surgical, Oncological and
Gastroenterological Sciences, University of Padua,
Padua, Italy
Background SerpinB3, a cysteine-proteases inhibitor upregulated in hepatocellular carcinoma (HCC), proposed as
biomarker of liver carcinogenesis and to stimulate epithelial-tomesenchymal transition (EMT) and increased invasiveness in liver
cancer cells, has been suggested to be up-regulated by hypoxia
through a HIF2␣-dependent mechanism.
Aims To investigate the selectivity of HIF2␣-related upregulation of SerpinB3 and the in vivo prognostic relevance of this
relationships.
e42
Methods Selectivity ofthe relationships between hypoxia,
HIF2␣ and SerpinB3 versus SerpinB4 polymorphic isoform expression has been investigated in control HepG2 cells or in HepG2 cells
stably transfected to overexpress HIF1␣ or HIF2␣ by employing
cell and molecular biology techniques. HIF2␣ and SerpinB3 transcript levels were evaluated (quantitative real-time PCR) on frozen
HCC specimens obtained at the time of surgical resection from 67
patients with cirrhosis of different aetiology. A series of 18 paraffinincluded liver specimens from cirrhotic HCV patients carrying HCC
was also included in the study for immuno-histochemistry analysis.
Results Selectivity of SerpinB3 up-regulation by hypoxia via
HIF2␣ was established by the following experimental evidence:
i) HIF2␣ (not HIF1␣), binds to the Serpin B3 promoter (ChIP
assay); ii) only HepG2 overexpressing HIF2␣ show SerpinB3 upregulation; iii) hypoxic conditions do not affect expression of
SerpinB4. Immuno-histochemistry and transcript analysis, performed in human HCC specimens, revealed co-localization of the
two proteins in liver cancer cells and the existence of a positive
correlation (Spearman r coefficient 0,3533, p < 0.01) between HIF2␣ and SERPINB3 transcript levels. Moreover, the highest levels of
HIF-2␣ transcripts were found in specimens with the highest levels
of SerpinB3 transcripts obtained from the most aggressive subset of HCC cases, correlating with the highest rate of early tumour
recurrence.
Conclusions A positive correlation between HIF2␣ and SerpinB3 expression exists in human HCC specimens which is selective
for the SerpinB3 isoform and has prognostic implications.
T-50
THE ACTIVATION OF NF-KB, PREGNANE X
RECEPTOR, AND CONSTITUTIVE ANDROSTANE
RECEPTOR IS MODULATED BY THE DEGREE OF
CHOLESTASIS
D. Gabbia 1 , T. Baldovin 2 , R. Lazzari 2 , C. Mescoli 3 ,
L. Albertoni 3 , V. Baldo 2 , A. Floreani 4 ,
S. De Martin 1
1 Department of Pharmaceutical and
Pharmacological Sciences, University of Padua,
Padua, Italy
Padua, Padua, Italy
Padua, Italy
Introduction: NF-kB is known to be activated in the early stages
of cholestasis, acting a reduction of liver injury. Pregnane X Recep-
tor (PXR) and Constitutive Androstane Receptor (CAR) are two
nuclear receptors (NRs) that regulate bile acid metabolism and
transport. A mutual negative crosstalk between NF-kB and NRs has
been reported, but conflicting data are available on the impact of
their relationship in cholestasis.
Aim: To analyse the changes in activation of NF-kB, PXR, and
CAR in an experimental model of cholestasis.
Methods: Cholestasis was induced in 8 male Wistar rats by
bile duct ligation; 4 sham-operated rats were used as controls. The
degree of cholestasis was defined on the basis of histologic examination of liver sections and serum concentration of albumin, ALT,
GGT, bilirubin. Activation of PXR, CAR and NF-kB was evaluated by
Western blot analysis on nuclear liver fractions.
Results: A 2-fold increase in activation of NF-kB was observed in
early stage of cholestasis (p < 0.05 with respect to Sham), whereas
the nuclear translocation of NF-kB was completely abolished in
the late stage (p < 0.001). A significant increase in PXR activation
was observed in late stage of cholestasis compared with both early
stages and controls (2- and 1.5-fold, p < 0.01 and 0.05, respectively).
CAR nuclear expression was 3.4-fold higher in early cholestatic rats
than in controls (p < 0.001), whereas CAR activation was virtually
abolished when cholestasis became severe (p < 0.05 and p < 0.001
vs. sham and early-stage cholestatic rats, respectively).
Conclusions: We provided new evidences of the strict relationship between NRs and NF-kB activation, suggesting a different
effect of NF-kB on PXR and CAR activation, which is dramatically
enhanced in severe cholestasis. Therefore, a different modulation
of the NF-kB pathway by acting on PXR and CAR according to the
degree of cholestasis may be considered as a new therapeutic strategy.
Liver – Annual Meeting 2015: Poster sessions – FRIDAY
F-01
UTILITY BASED CRITERIA TO SELECT PATIENTS
WITH HEPATOCELLULAR CARCINOMA FOR
LIVER TRANSPLANTATION: A MULTICENTRE
COHORT STUDY
A. Vitale 1 , F. Farinati 2 , P. Burra 2 , F. Trevisani 3 ,
E.G. Giannini 4 , G. Spolverato 1 ,
U. Cillo 1 , for the Italian Liver Cancer (ITA.LI.CA) group
1
Dipartimento di Chirurgia Generale e Trapianto
d’Organo, Unità di Chirurgia Epatobiliare e Trapianti
Epatici, Università di Padova, Padova, Italy
2 Divisione di Gastroenterologia, Azienda Università
di Padova, Padova, Italy
3 Dipartimento di Scienze Mediche e Chirurgiche,
Unità di Semeiotica Medica, Alma Mater
Studiorum–Università di Bologna, Bologna, Italy
4 Dipartimento di Medicina Interna, Unità di
Gastroenterologia, Università di Genova, Genova,
Genova, Italy
Background. The lifetime utility of liver transplantation (LT) in
patients with hepatocellular carcinoma is still controversial. The
aim of this study was to understand when LT is cost-effective for
HCC patients in order to suggest new potential transplant selection
criteria.
Methods. The overall design of this study involved a real cohort
of transplantable Italian HCC patients (n = 2419 selected from the
ITA.LI.CA database) undergoing non-transplant therapies for whom
a survival analysis and calculation of direct costs of therapies were
performed, and then a Markov model to calculate the survival benefit and cost-utility of LT over non-LT therapies. Post LT survival was
calculated using the ␣-fetoprotein (AFP) model based on AFP values
and radiological size and number of nodules. Primary endpoint was
Net Health Benefit (NHB), defined as LT survival benefit in quality
adjusted life years (QALYs) minus incremental costs ($)/willingness to pay ($32,946, corresponding to the Italian Gross Domestic
Product).
Results. We developed a prognostically accurate survival model
predicting non-LT survival, based on AFP model variables, Child
Pugh classes, and type of alternative therapy (resection, locoregional, or best supportive care). The calculated median cost of
non-LT therapies per patient was $21,670. The Monte Carlo simulation showed that in patients with resectable child A HCC, the NHB
1590-8658/$36.00
of LT was always negative, independently from the AFP model values, while in unresectable HCC patients, the NHB of LT was positive
for AFP model values ≤ 4, and negative for values > 4.
Conclusion. LT proved to be cost-effective in HCC patients with
unresectable tumor and AFP model values ≤ 4.
F-02
STIMULATION OF NUCLEAR RECEPTOR PPAR-␥
LIMITS NF-KB-DEPENDENT INFLAMMATION IN
CYSTIC FIBROSIS BILIARY EPITHELIUM
R. Scirpo 1,2 , R. Fiorotto 1 , A. Villani 1 , L. Fabris 3 ,
M. Strazzabosco 1,2
1
Department of Internal Medicine, Yale University,
New Haven, USA
2 Department of Surgery and Traslational Medicine,
University of Milano-Bicocca, Monza, Italy
3 Department of Surgical Oncological and
Gastroenterological Science, University of Padua,
Padua, Italy
Background. Cystic Fibrosis-associated liver disease (CFLD) is
a chronic cholangiopathy that negatively affects life of Cystic
Fibrosis patients. We have recently shown that altered TLR/NFkB-dependent biliary innate immune mechanisms contribute to
the pathogenesis of CFLD, and may represent novel therapeutic
targets for CFLD. Nuclear receptors (NRs) are ligand-dependent
transcription factors that, by controlling gene expression, regulate
several intracellular functions. Selected classes of NRs, including peroxisome proliferator-activated receptor ␥ (PPAR-␥), also
counter-regulate inflammation in several tissues, by cross-talking
with TLR-dependent signaling pathways. We hypothesized that
pharmacologic activation of nuclear receptor PPAR-␥ would inhibit
the TLR/NF-kB-dependent inflammation in Cftr-defective biliary
epithelium.
Results. The gene and protein expression of PPAR-␥ is increased
in CF cholangiocytes respect to WT cells, but does not correlate
with a higher expression of PPAR-␥ target genes (Acaa1b, Angptl4
and Hmgcs2), indicating that the receptor is not more active. Consistently, Cftr-KO cells show a decreased production of PPAR-␥
endogenous ligands that might limit a proper activation of the
receptor. Treatment with LPS caused a higher NF-kB activation and
e44
NF-kB-dependent cytokine secretion in Cftr-KO cells, compared to
controls. In the presence of PPAR-␥ agonists, the activation of NF-kB
and the secretion of proinflammatory cytokines LIX, MCP-1, MIP2 and G-CSF were significantly inhibited at baseline and after LPS
challenge. Modulation of NF-kB-dependent inflammation by PPAR␥ resulted from the induction of the NF-kB negative regulator IkB␣.
Finally, stimulation of PPAR-␥ in vivo significantly attenuated biliary damage and inflammation in Cftr-KO mice after DSS-induced
portal endotoxemia.
Conclusions: Our studies unravel a novel mechanism of regulation of biliary epithelium innate immunity and suggest that
stimulation of PPAR-␥ signaling by synthetic agonists may represent a valuable therapeutic option to limit biliary inflammation
in CFLD. We expect that they might be applicable to other organs
affected by CF and to the treatment of other inflammatory cholangiopathies.
F-03
DEEP-SEQUENCING ANALYSIS DEMONSTRATES
THE PERSISTENCE OF PRE-TRANSPLANT HCV
DOMINANT VARIANTS WITHIN A MORE
HOMOGENEOUS QUASISPECIES AFTER LIVER
TRANSPLANTATION (LT) IN CHOLESTATIC
HEPATITIS C PATIENTS
M. Gambato 1,2 , J. Gregori 3,4 , J. Quer 3 ,
N. Caro-Pérez 1 , G. Crespo 1 , M. Navasa 1 ,
S. Pérez-del-Pulgar 1 , X. Forns 1
1
Liver Unit, Hospital Clínic, IDIBAPS, Ciberehd,
Barcelona, Spain
3 Liver Unit, Vall d’Hebron Institut de
Recerca-Hospital Universitari Vall d’Hebron,
CIBERehd, Barcelona, Spain
4 Roche Diagnostics SL. Sant Cugat del Vallès,
Barcelona, Spain
Introduction. Little is known about the pathogenic mechanisms
implicated in cholestatic hepatitis C (CH). Host failure to mount
specific T-cell responses against HCV has been implicated in the
pathogenesis. However, a direct role of HCV is not well established.
Aim. The aim of this study was to analyze, for the first time,
the HCV quasispecies evolution in patients with CH (compared to
a control group) by ultra-deep pyrosequencing (UDPS).
Materials and Methods. 13 HCV-infected liver transplant recipients with CH and 10 with mild recurrence (control group) have
been analyzed so far. A serum sample at the time of LT and a second sample obtained 1-3 months after LT were analyzed for each
patient. UDPS of a NS5B fragment (339 nucleotides) was performed
using the 454 GS Junior platform.
Results. Donor age (p = 0.007), recipient age (p = 0.022) and viral
load after LT (p < 0.001) were significantly higher in patients with
CH compared to controls. The mean number of reads per sample was 6146 (range 2491-25820). NS5B quasispecies complexity
before LT was similar in CH and control group (p = 0.396). On the
contrary, genetic diversity and mutation frequency of NS5B quasispecies decreased significantly after LT in patients with CH (p = 0.013
and p = 0.013), but not in the control group (p = 0.069 and p = 0.327).
Interestingly, phylogenetic analysis showed that the major HCV
variant pre-LT successfully propagated and remained as the major
variant after LT in 64% of patients with CH, but only in 12% of controls (p = 0.026).
Conclusions. A marked homogenization of HCV quasispecies
was demonstrated in HCV-infected patients who developed CH
after LT. In addition, in most CH patients, the major HCV strain
before LT remained as dominant after LT. The latter suggests that, in
the context of immunosuppresion, the propagation of more fitted
viral strains could play a role in the pathogenesis of CH.
F-04
OVERALL SURVIVAL IN INTERMEDIATE-STAGE
HEPATOCELLULAR CARCINOMA (HCC) PATIENTS
AFTER FIRST TRANSARTERIAL
CHEMOEMBOLIZATION (TACE): PROPOSAL OF A
NEW SCORING SYSTEM
R. Sacco 1 , B. Ginanni 3 , V. Mismas 1 , G. Masi 2 ,
P. De Simone 2 , A. Romano 1 , G. Bresci 1 ,
C. Bartolozzi 3 , I. Bargellini 3
1 Department of Gastroenterology, Pisa University
Hospital, Pisa, Italy
2 Department of Oncology, Pisa University Hospital,
Pisa, Italy
3 Department of Radiology, Pisa University Hospital,
Pisa, Italy
Introduction TACE is the standard treatment for patients with
intermediate-stage HCC. However, prognostic factors for survival
after the first TACE cycle remain unclear.
Aims We correlated pre-treatment characteristics and response
to therapy with overall survival (OS) and time to progression (TTP),
in order to propose a scoring system aimed at facilitating clinical
decision after the first TACE.
Patients and methods We retrospectively analyzed 149 patients
with intermediate-stage HCC who received ≥1 TACE cycle(s).
Univariate and multivariate analysis were used to correlate pretreatment characteristics and response to TACE with OS and TTP.
Predictive factors were used to define a score for each patient.
Results Median OS was 23 (95% CI 11.5-27) months, and median
TTP was 11 months (7-11). Complete response was reported in 63
patients (42.3%) and partial response (PR) in 71 (47.7%).
Age >65 years (HR 1.77; 95% CI: 1.18-2.67), ascites (HR 2.44; 95%
CI 1.32-4.29), total diameter of nodules >61 mm (HR: 1.96; 95% CI
1.28-3.08) and response at 1 month (HR 1.70; 95% CI 1.30-2.20)
were predictors of survival and were used to build the scoring system. Age >65 years, ascites, total diameter of nodules >1 accounted
for one score point; PR, stable disease and progression of disease
accounted for 1, 2 and 3 score points, respectively. The score points
were summed to calculate a single score for each patient.
Patients with score 0-1 had a longer OS (57.8 months) and TTP
(12.7 months) than those with score 2-3 (21.1 and 8.2 months) or
score 4-6 (8.0 and 6.3 months) (p < 0.001 for both comparisons).
Conclusions This scoring system may allow the identification of
three groups of patients with different prognosis after a first TACE
cycle and may therefore help guide clinical decisions, in particular
whether continuing TACE therapy after a first cycle or moving to
different therapies.
F-05
TREATMENT OF ADVANCED HEPATOCELLULAR
CARCINOMA (HCC): A SINGLE CENTRE COST
ANALYSIS OF YTTRIUM 90 TRANS-ARTERIAL
RADIO-EMBOLIZATION (TARE) VERSUS
SORAFENIB
M.G. Lucà 1 , M. De Giorgio 1 , G. Magini 1 ,
A. Tortora 4 , L. Sangiovanni 1 , G. Gaffuri 1 ,
A. Baldan 1 , G. Virotta 3 , R. Nani 2 , S. Fagiuoli 1
1
Department of Internal Medicine, Papa Giovanni
2 Department of Radiology, Papa Giovanni XXIII
Hospital, Bergamo, Italy
3 Department of Nuclear Medicine, Papa Giovanni
4 Division of Internal Medicine and Gatroenterolgy,
Department of Internal medicine, A. Gemelli
INTRODUCTION: In a selected population of advanced HCC
patients TARE seems to give promising results.
AIM: To compare a single Centre costs for Sorafenib and TARE
METHODS: 166 consecutive advanced HCC patients (M 86%,
median age 67 yrs) treated with Sorafenib between June 2009 and
June 2014 (Group SOR) and 19 (M 84%, median age 64 yrs, BCLC B
31,5%) treated with TARE between June 2011 and June 2014 (Group
TARE) were compared.
Patients were grouped for treatment time (Group SOR 1: < 2
months of treatment; Group SOR 2: > 2 months of treatment).
Group SOR 1 drug expenses were not further considered according with the agreed “pay for result policy”. In group SOR 2, 24 pts
(group SOR 3) presented monolobar disease and no methastasis
(BCLC B 54%) so as to be potentially treated with TARE. Sorafenib
cost was calculated according with the total effective caps intake
for each patients (28,7 Euro/caps).
TARE costs included Yttrium-90 and hospitalization expense
(17.761 Euro/patient).
RESULTS: Median time of treatment in Group SOR 3 was 272
days (154-994) with a median intake of 2,8 tablets/day. All the
patients in the group TARE were treated with a single treatment
session. Median follow up time was respectively 476 days (1541518) for the Group SOR 3 and 266 days (47-867) for the Group
TARE.
Survivals at 12, 18, 24 and 36 months were respectively 66,7%,
37,5%, 24,3% and 19.4% for the Group SOR 3 and 64.1%, 64.1%, 64.1%
and 32% for the Group TARE (Log Rank p = 0.446). The GLOBAL treat-
e45
ment costs for Group SOR 3 and for Group TARE were D 671.809,6
and D 337.459,00 respectively (median D 27992/pt in SOR3 vs D
17761/pt in TARE; p = 0.028).
CONCLUSION: TARE treatment in advanced HCC may reduce
costs with survivals not significantly different from Sorafenib.
F-06
CORONARY MICROVASCULAR DYSFUNCTION IN
PATIENTS WITH PRIMARY BILIARY CIRRHOSIS
WITHOUT METABOLIC SYNDROME: A HINT FOR
THEIR INCREASED CARDIOVASCULAR RISK
N. Cazzagon 1 , C. Dal Lin 2 , G. Famoso 2 ,
I. Franceschet 1 , A. Floreani 1 , F. Tona 2
2 Department of Cardiac, Thoracic and Vascular
Sciences, University of Padua, Padua, Italy
Background: Primary biliary cirrhosis (PBC) is characterized
by a long natural history and a low incidence of cardiovascular
events despite high serum cholesterol levels. Metabolic syndrome
may increase the cardiovascular risk, however. Coronary flow
reserve (CFR) is widely used to examine the integrity of coronary
microvascular circulation and it is well recognized as a predictor of
cardiovascular outcome.
Aim: To evaluate the risk of coronary microvascular dysfunction
in patients with PBC without metabolic syndrome.
Methods: 29 PBC patients (27 F, aged 56 ± 11 years) without
clinical evidence of heart disease and without metabolic syndrome, and 29 sex- and age-matched healthy controls underwent
CFR by transthoracic Doppler echocardiography (TDE). Coronary
flow velocity in the left anterior descending coronary artery was
detected by TDE at rest and during iv. adenosine infusion. CFR was
the ratio of hyperaemic diastolic flow velocity (DFV) to resting DFV.
Results: The median time between the onset of symptoms and
CFR assessment was 7 years (interquartile range 3-10.5 years). In
PBC patients CFR was significantly lower than in controls (3.1 ± 0.3
vs 3.8 ± 0.4, p < 0.0002) (Fig. A). CFR was inversely related to the
time from diagnosis (r = -0.389, p = 0.03) (Fig. B) and age (r = -0.370,
p = 0.04), and positively correlated with TSH (r = 0.411, p = 0.03). No
relationship was found between CFR and LDL-cholesterol.
Conclusions: CFR is impaired in PBC patients and is correlated
with the length of disease, suggesting a negative effect of PBC on
coronary microcirculation that may contribute to the increased cardiovascular risk, even in patients without metabolic syndrome.
e46
F-07
F-08
THE TM6SF2 E167 K VARIANT IS AN
INDEPENDENT PREDICTOR OF SEVERE LIVER
STEATOSIS IN CHRONIC HEPATITIS C
METABOLIC SYNDROME AFTER LIVER
TRANSPLANTATION: HAVE RISK FACTORS
CHANGED OVER TIME?
N. Coppola 1 , R. Zampino 2 , G. Cirillo 3 ,
M. Stanzione 4 , M. Macera 1 , A. Boemio 2 ,
A. Grandone 3 , M. Pisaturo 1 , A. Marrone 2 ,
L.E. Adinolfi 2 , E. Sagnelli 1 ,
E. Miraglia del Giudice 3
E. Vanni 1 , R. Ibrahim Kamal Jouness 1 ,
S. Mirabella 2 , A. Marengo 1 , A. Milan 3 , C. Rosso 1 ,
V. Boano 1 , E. Mosso 1 , C. Di Stefano 3 , E. Nada 2 ,
M. Rizzetto 1 , M. Salizzoni 2 , R. Romagnoli 2 ,
E. Bugianesi 1
Naples, Naples, ITALY
2 Internal Medicine and Hepatology, Second
University of Naples, Naples, ITALY
3 Department of Pediatrics, Second University of
Naples, Naples, ITALY
4 Department of Clinical and Experimental Medicine
and Surgery, Second University of Naples, Naples,
ITALY
Gastroenterology, University of Turin, Turin, Italy
2 Liver Transplant Center, University of Turin, Turin,
Italy
Internal Medicine, University of Turin, Turin, Italy
Background: A common non-synonymous polymorphism,
E167 K, in transmembrane 6 superfamily member 2 (TM6SF2) gene
has been recently associated with an increased hepatic triglyceride
content, dyslipidemia and liver fibrosis in NAFLD patients.
Aim: We investigated on a possible associations between
TM6SF2 variants and degree of liver lesions in chronic hepatitis
C.
Methods: 148 consecutive patients with biopsy proven antiHCV/HCV-RNA-positive chronic hepatitis, naive for antiviral
therapy, were genotyped for TM6SF2 E167 K and PNAPL3 I148 M
variants by a TaqMan Assay. These patients aged 50.9 ± 12.8, 54.7%
were males, 14.9% with a history of alcohol abuse, 62.2% with HCV
genotype 1 and 9.3% with genotype 3.
Results: 130 (87.8%) patients were TM6SF2 167E homozygotes
and 18 (12.2%) heterozygotes for the E167 K variant. The prevalences of patients with the PNAPL3 I/I, I/M or M/M were 26.3%,
59.1% and 14.6%, respectively. Patients with different TM6SF2
variants did not show significant differences in demographics, biochemical data including cholesterol and triglyceride, HCV load,
prevalence of patients with HCV genotype 3 and distribution of
PNPLA3 variants. As compared with the 130 homozygotes for
TM6SF2 167E allele, the 18 patients with TM6SF2 E167 K variant showed a higher degree (scale from 1 to 4) of liver steatosis
(1.9 ± 1.3 vs. 1.1 ± 1.1, p = 0.02) and a higher prevalence of cases
with a steatosis score ≥ 3 (33.3% vs. 12.3%, p = 0.02); no difference
in necroinflammation nor in fibrosis scores was observed. A general linear model identified as independent predictors of steatosis
the TM6SF2 E167 K, the PNAPL3 M148 M variants and the waist
circumference (p = 0.0376, p = 0.0069 and p = 0.0273, respectively).
The effect of the PNAPL3 M/M variant on liver steatosis looks significantly enhanced when associated with the TM6SF2 E/K variant
(OR 11.4, CI 95% 1.06-134, p = 0.04).
Conclusions: This study is the first demonstration that TM6SF2
E167 K variant is an independent predictor of severe liver steatosis
in chronic hepatitis C.
Background and Aims: Post-Transplant Metabolic Syndrome
(PTMS) is common among liver transplantation (OLT) recipients
The aim of this study is the definition of the factors contributing to PTMS development with particular regard to changes in the
immunosuppressive regimens.
Methods: Three hundred and ninety-three patients who underwent OLT at Turin Liver Transplant Center were prospectively
enrolled (group A: pre-mTOR inhibitors - OLT 1998-2003, n = 167;
group B: m-TOR inhibitors - OLT 2008-2012, n = 226). Overall the
median follow-up was 60 months (1-204), 144 (83-204) in group A
and 48 months (1-60) in group B. Clinical, laboratory parameters,
prescribed medications, were recorded every 6-12 months. Clinical
features of the donors and histological data of the allograft were collected. Multivariate analysis were performed to identify risk factors
associated with PTMS development.
Results: Clinical features and OLT indications were comparable between the two groups. Cryptogenic cirrhosis was an OLT
indication in 2.8% (11/393) without temporal changes. Overall preOLT MS was found in 1.8% (7/393) subjects, without temporal
changes, while PTMS in 24.9% (98/393; group A 37.9%, group B
22.6%, p = 0.003). Overall multivariate analysis identified pre-OLT
hyperglycemia and post-OLT weight gain as predictive risk factors for PTMS. However, independent risk factors were different
in the two groups: post-OLT weight gain (OR 1.53, 95% CI 1.261.85, p = 0.0001) and post-OLT US liver steatosis (OR 4.12, 95% CI
1.48-11.4, p = 0.006) in group A; post-OLT weight gain (OR 1.36, 95%
CI 1.17-1.57, p = 0.0001), pre-OLT hyperglycemia (OR 3.57, 95% CI
1.28-9.88, p = 0.014), macrovacuolar steatosis ≥20% in the allograft
(OR 4.74, 95% CI 1.23-18.27, p = 0.024) and treatment with m-TOR
inhibitors (OR 17.43, 95% CI 3.36-90.39, p = 0.001) in group B.
Conclusions: Overall, host-related factors contribute to PTMS
development, but the introduction of mTOR inhibitors is an
additional risk factor particularly when suboptimal livers are transplanted.
F-09
F-10
CLINICAL EVALUATION OF CIRCULATING
MICRORNAS AS POTENTIAL BIOMARKERS OF
HEPATOCELLULAR CARCINOMA IN PATIENTS
WITH CHRONIC HBV INFECTION
THE TWO THIENO-TRIAZOLODIAZEPINES WEB
2086 AND WEB 2170 BLOCK
HEPATOCARCINOMA PROGRESSION BY
HSP-DEPENDENT CLIENT KINOMA PROTEIN
UBIQUITINATION
G.P. Caviglia 1 , M.L. Abate 1 , E. Petrini 2 , S. Gaia 2 ,
P. Manzini 3 , P. Carucci 2 , M. Rizzetto 1,2 ,
A. Smedile 1,2
1
Department of Medical Sciences, University of
Turin, Turin, Italy
2 Department of Gastroenterology and Hepatology,
Città della Salute e della Scienza Hospital, Turin, Italy
3 Blood Bank, Città della Salute e della Scienza
Hospital, Turin, Italy
Introduction. Several studies showed that aberrant miRNA
expression is associated with development and progression of
hepatocellular carcinoma (HCC).
Aim. To examine whether some commonly deregulated miRNAs
(miR-122, miR-21, miR-221 and miR-16) in HBV-related HCC may
serve as diagnostic markers.
Materials and Methods. Serum expression of miR-122, miR21, miR-221 and miR-16 was evaluated by real-time quantitative
RT-PCR in 33 patients with HBV-related HCC (61 ± 10 years;
F/M = 4/29), 30 patients with HBV-related cirrhosis (53 ± 11 years;
F/M = 11/19) and 27 blood donors as healthy controls (54 ± 8 years;
F/M = 9/18).
Results. Median levels of miR-16, miR-122 and miR-221 were
significantly different in patients with HCC or cirrhosis compared to
healthy controls (p < 0.001) whereas, only miR-122 levels differed
in patients with HCC from cirrhotic patients (p = 0.024). miR-122
levels were significantly higher in patients with multifocal HCC
than in patients with a single lesion (p = 0.024). Expression levels
of mir-21 were similar in the 3 groups. Area under the curve (AUC)
analysis showed that serum levels of miR-122, miR-122 + miR221, miR-122 + miR-16 and miR-122 + miR-221 + miR-16, are able
to differentiate patients with HCC from patients with cirrhosis
(AUC = 0.675; AUC = 0.704; AUC = 0.681; AUC = 0.703, respectively).
Moreover, miR-16, miR-122 and miR-221, alone or in combination, were able to discriminate patients with HCC or cirrhosis from
healthy controls (AUC > 0.9). In addition, a correlation between
miR-122 levels and HCC nodules number (R = 0.390; p = 0.036), and
between miR-16 and miR-122 levels, and ALT values (R = -0.464,
p = 0.034; R = 0.449, p = 0.536, respectively) was found.
Conclusions. Among the four microRNAs analyzed, only miR122 significantly discriminates patients with HCC from cirrhotic
patients and patients with HCC or cirrhosis from controls. miR122 appears to reflect liver necro-inflammation, since we observed
a positive correlation with ALT levels. Moreover, the correlation
between miR-122 expression levels and HCC with multi-nodules,
suggests the possible use of this miRNA for tumor stadiation. Nevertheless, miR-122 AUC values were not sufficiently high for HCC
screening purposes.
e47
E. Ceni 1 , C. Malentacchi 1 , M. Tarocchi 1 ,
G. Marroncini 2 , S. Polvani 1 , T. Mello 1 ,
S. Tempesti 1 , A. Galli 1,2
1 Experimental and Clinical Biomedical Sciences
Mario Serio, University of Florence, Florence, Italy
2 FiorGen Foundation, Florence, Italy
Introduction: L’HCC is one of the most common malignancies
worldwide. So far there is no effective chemotherapeutic treatment for HCC and the prognosis of advance stage remains poor.
The thieno-triazolodiazepine are well known anti-inflammatory
drugs that acts as PAF-receptor antagonists. Recently an antineoplastic pleiotropic effect of this molecules have been shown,
where treatment with thieno-triazolodiazepine (WEB2086 and
WEB2170) induce differentiation, growth arrest and apoptosis in
murine and human leukemia cells. The effects of WEBs on HCC
remain untested. Aim of this study was to investigate the antitumor efficacy of the WEBs in vitro and in vivo models of HCC.
Material and Methods Results: WEBs were able to reduce proliferation of cancer cell lines (HepG2, Hep3B, HuH7 and Hepa 1-6)
as assessed by thymidine incorporation and eliminates the ability of these cells to form colonies in soft agar that was associated
with cell cycle arrest, apoptosis, and senescence. In addition, WEBs
impair hepatoma cell migration in a Wound Healing assay and
chemoinvasion on Matrigel. WEBs administration in HBV transgenic mice reduces the number and the dimension of tumor masses.
Antitumorigenic effects in mice were also confirmed in a HepG2
xenograft model of HCC. A 2D-DIGE proteome study highlights that
the WEB2086 inhibits the binding processes in particular inhibits
protein binding of ATP and GTP and the abrogation of this cellular
function results kinoma inhibition. In addition 2D-DIGE analysis showed that WEBs down-regulate chaperone proteins such as
hsp90 and hsp70 that are involved in protein refolding, an important mechanism in tumor cell resistance. The down-regulation
dell’Hsp90 lead to failure of “clients” protein refolding and mediate
their degradation.
Conclusions: The thieno-triazolodiazepines are able to reduce
HCC progression in human and murine HCC models blocking cancer
cell proliferation and migration and inducing apoptosis and senescence probably whit a mechanism that involve kinome inhibition.
e48
F-11
CLINICAL RELEVANCE OF NEXT GENERATION
SEQUENCING ON BASELINE DETECTION OF
MINORITY RESISTANCE ASSOCIATED VARIANTS
IN HCV-1 PATIENTS TREATED WITH PROTEASE
INHIBITORS
D. Armenia 1 , L. Carioti 1 , V.C. Di Maio 1 ,
M.C. Bellocchi 1 , D. Di Paolo 2 , F. Guerrieri 3 ,
L. Calvo 4 , V. Cento 1 , M. Tontodonati 5 ,
V. Micheli 6 , F. De Leonardis 2 , M. Aragri 1 ,
E. Polilli 7 , A. Manunta 8 , C. Magni 6 ,
F.P. Antonucci 1 , F. De Luca 1 , C. Sarrecchia 9 ,
A. Bertoli 1 , I. Lenci 2 , S. Francioso 2 ,
M.M. Santoro 1 , J. Vecchiet 5 , S. Marenco 10 ,
A. Picciotto 10 , L. Nosotti 11 , F. Morisco 12 ,
S. Bruno 13 , M. Puoti 14 , S. Babudieri 8 , M.S. Mura 8 ,
M. Andreoni 9 , G. Rizzardini 6 , G. Parruti 7 ,
M. Levrero 4 , M. Angelico 2 , C.F. Perno 1 ,
F. Ceccherini-Silberstein 1
1 Department of Experimental Medicine and Surgery,
University of Rome “Tor Vergata”, Rome, Italy
2 Hepatology Unit, University Hospital of Rome “Tor
Vergata”, Rome, Italy
3 Center for Life Nano Science, CNLS@SAPIENZA, IT
4 La Sapienza” University, Rome, Italy
5 Infectious Disease Clinic, Chieti, Italy
6 Hospital Sacco of Milan, Milan, Italy
7 Infectious Disease Unit, Pescara General Hospital,
Pescara, Italy
8 Department of Clinical and Experimental Medicine,
University of Sassari, Sassari, Italy
9 Infectious Disease, University Hospital of Rome “Tor
Vergata”, Rome, Italy
10 S. Martino University Hospital, Genoa, Italy
11 Hepatology Unit, National Institute of Health,
Migration and Poverty, Rome, Italy
12 University “Federico II”, Naples, Italy
13 Internal Medicine, Gastroenterology and
Hepatology, Azienda Ospedaliera Fatebenefratelli e
Oftalmico, Milan, Italy
14 Hospital Niguarda Ca’Granda, Milan, Italy
Background and Aims: This study aims to evaluate the
clinical relevance of next-generation-sequencing on the baseline detection of minority resistance-associated-variants (RAVs)
in chronic HCV-1 infected patients treated with telaprevir/
boceprevir + pegIFN/ribavirin.
Methods: NS3-protease sequences of 25 selected patients
(18 virological-failures and 7 responders) treated with telaprevir (N = 20) or boceprevir (N = 5) + pegIFN/ribavirin were analyzed.
Detection of NS3-RAVs (V36ALM/T54AS/V55A/Q80 K/R155KT/
A156STV/V170A) was performed by Sanger-sequencing, 454junior-pyrosequencing (UDPS) and MiSeq-illumina (MiSeq).
UDPS sequences were analyzed by a home-made-pipeline
(>3,000 sequences/patient; cut-off > 0.1%); Miseq sequences were
analyzed by VirVarSeq (>40,000 sequences/patient; cut-off ≥ 0.3%).
Results: At baseline, 5/25 patients (20%) presented NS3-RAVs
by Sanger, UDPS and MiSeq (Q80K = 3; T54S = 1; V36L + Q80K = 1,
prevalence >98%). Of them, 4 patients experienced virologicalfailure and 1 (with Q80 K) achieved sustained-virological-response
(SVR). Additional baseline minority-RAVs (Q80 K/V170A) were
found by both UDPS and MiSeq in 2 patients, all with prevalence < 2.6%. However, their presence was not directly associated
with virological-failure. Indeed, 1 previous-null-responder with
baseline minority Q80 K failed telaprevir-triple-therapy with different RAVs (V36AM + R155K + T156ST). The other patient, previous
partial-responder, with baseline minority V170A achieved SVR.
By using MiSeq, analyzing a larger number of sequences
(median[IQR]:843,492[342,499-1,006,108]), a high number of NS3RAVs (N = 43,cut-off ≥0.3%) were detected in 19/25 patients (76%).
In particular, V55A was the most prevalent RAV detected (52%),
followed by V36A (36%), Q80 K (28%), and T54A (8%). The presence
of these mutations at baseline was similar between responders
and virological-failures. Interestingly, a unique HCV-1a infected
patient previous-non-responder with 10 baseline minorityRAVs (V36A:0.41%; T54S:0.33%; V55A:0.47%, Q80 K:0.36%;
R155 K:0.55%;
R155 T:0.59%;
A156S:1.19%;
A156 T:0.64%;
A156 V:0.50%, V170A:0.57%, with a median[IQR] mutational-load
of 19,567[15,870-21,987] IU/ml) failed telaprevir-triple-therapy,
with A156 T (week-2) and later with V36M + R155 K (week-10).
Conclusions: Both UDPS and MiSeq detected baseline RAVs at
very low frequency. However, these variants were not necessarily
detected at virological-failure. Further investigations are needed to
clarify the clinical relevance of minority RAVs at frequency below
1%.
F-12
CHRONIC INTERMITTENT HYPOXIA IS
ASSOCIATED WITH LIVER DAMAGE AND
ATHEROSCLEROSIS IN PATIENTS WITH
NON-ALCOHOLIC FATTY LIVER DISEASE
S. Petta 1 , O. Marrone 2 , D. Torres 3 ,
M. Buttacavoli 4 , C. Cammà 1 , V. Di Marco 1 ,
A. Licata 1 , A. Lo Bue 2 , G. Parrinello 3 , A. Pinto 3 ,
A. Selvaggio 2 , A. Tuttolomondo 3 , A. Craxì 1 ,
M. Bonsignore 4
1 Sezione di Gastroenterologia, DiBiMIS, University of
2 National Research Council, Institute of Biomedicine
and Molecular Immunology, Palermo, Italy
3 Sezione di Medicina Interna, DiBiMIS,University of
4 Sezione di Pneumologia, DiBiMIS,University of
Background: Obstructive sleep apnea syndrome(OSAS) has
been reported as a new risk factor for metabolic disturbances,
including cardiovascular alterations. Growing data are also available among bariatric nonalcoholic fatty liver disease(NAFLD)
patients on the impact of obstructive sleep apnea syndrome OSAS
on liver damage in NAFLD. We assessed whether OSAS is associated
with severity of liver fibrosis and carotid atherosclerosis in NAFLD
patients without morbid obesity.
Methods: 126 consecutive biopsy-proven(Kleiner score) NAFLD
patients assessed for anthropometric, biochemical, and metabolic
features underwent ultrasonographic carotid assessment and STOP
BANG questionnaire for estimate of low or high OSAS risk. A carotid
plaque was defined as a focal thickening > 1.3 mm at the level of
either common and internal carotid arteries or bifurcations. 50
patients accepted to perform nocturnal cardiorespiratory polygraphy, and OSAS was defined if the as an apnea/hypopnea index AHI
index was≥5.
Results: The prevalence of high OSAS risk was similar in patients
without and with polygraphy(76% vs 68%,p = 0.17). Among these
last subjects who underwent polygraphy 50% had OSAS. The prevalence of an OSAS was significantly higher in patients with, than in
those without, F2-F4 fibrosis compared to those without(72% vs
44%, respectively; p = 0.04). After correction for confounders, significant fibrosis was associated with S02 mean levels≤95% mean
oxyhemoglobin saturation(SaO2) levels < 95%(OR 3.21,95%C.I. 1.027.34;p = 0.04). Similarly, the prevalence of OSAS was slightly higher
in patients with, than in those without, carotid plaques compared
to(64% vs 40%;p = 0.08). After correction for confounders, the association between carotid plaques were associated with T90(time
spent with SaO2 < 90%) >1% was maintained(OR 6.30,95%C.I.1.0212.3;p = 0.01).
Conclusions: In NAFLD patients without morbid obesity OSAS
was highly prevalent and indexes of oxygen saturation were independent indicators of the severity of liver fibrosis and of carotid
atherosclerosis risk. These data, if further validated, could suggest
to look at for OSAS in all NAFLD patients, considering OSAS as a
potential additional therapeutic target for NAFLD.
F-13
IFNL4 POLYMORPHISMS PREDICT SUSTAINED
RESPONSE AND HBSAG CLEARANCE IN
INTERFERON TREATED HBEAG NEGATIVE
CHRONIC HEPATITIS B PATIENTS
E. Galmozzi 1 , P. Lampertico 1 , F. Facchetti 1 ,
F. Invernizzi 1 , G. Mangia 1 , M. Vigano 2 ,
R. Soffredini 1 , M. Colombo 1
1
2 Liver Unit, Ospedale San Giuseppe, University of
Milan, Milan, Italy
Introduction and Aim The recently identified interferon
lambda 4 (IFNL4) gene harbors the dinucleotide variant
rs368234815-TT/G, a genetic marker of outcome to IFN-based
therapy of HCV infection. The IFN␭-4 protein, which can be generated only by carriers of the rs368234815-G allele, is thought
to counteract IFN responsiveness by inducing a weak expression
of interferon-stimulated genes. Three nonsynonymous variants
of the IFNL4 gene (rs73555604, rs142981501 and rs117648444)
could affect the IFN␭-4 protein. We aimed to explore whether
IFNL4 polymorphisms impact on response to IFN-based treatment
in the setting of chronic hepatitis B (CHB).
Materials and Methods IFNL4 gene was sequenced by Sanger
method on genomic DNA extracted from whole blood of 126
HBeAg-negative CHB patients treated with either standard or
pegylated-IFN-␣ and followed-up for 11 years (range 1-17) posttreatment. Results Sustained response rates to IFN were not
significantly different between the 62 carriers of the rs368234815TT/TT (IFN␭-4 eliminating) genotype and the 64 carriers of the
rs368234815-G (IFN␭-4 generating) allele (31% vs 17%, p = 0.079).
Since the only nonsynonymous variant identified in our cohort, the
Pro70Ser rs117648444-C/T polymorphism, was exclusively associated with carriers of the IFN␭-4 generating allele, these 64 patients
were stratified into rs117648444-CC (IFNl-4 wild-type, n = 45) and
rs117648444-CT/TT (IFNl-4 mutated, n = 19) genotypes. Sustained
responses among IFN-␭4 eliminating (rs368234815-TT/TT) and
IFN-␭4 mutated (rs117648444-CT/TT) genotypes (n = 81) were significantly higher than those in the 45 IFN␭-4 wild-type subjects
(33.3% vs 9%, OR = 4.8, 95%CI 1.6-15.0, p = 0.006). Yet, in the multivariate analysis the combination of IFN␭-4 eliminating and IFN␭-4
mutated genotypes independently predict both sustained response
to interferon (OR = 5.33, 95%CI 1.7-16.8, p = 0.004) and off treatment
e49
HBsAg clearance (HR = 4.3, 95%CI 1.5-12.3, p = 0.007). Pretreatment
serum HBV-DNA was the only other independent predictor of
HBsAg loss (HR = 0.61, 95%CI 0.43-0.87, p = 0.007).
Conclusions IFNL4 polymorphisms independently predict sustained response to interferon and off treatment HBsAg clearance in
HBeAg negative CHB patients.
F-14
NON-INFECTIOUS CO-MORBIDITIES IN HIV
PATIENTS CO-INFECTED WITH HEPATITIS
VIRUSES: AN ANALYSIS FROM THE CALABRHIV
STUDY GROUP
M.C. Postorino 1 , F. Luciani 2 , C. Mangano 3 ,
M.S. Carpentieri 3 , P. Scerbo 4 , A. Priamo 4 ,
G. Berardelli 5 , R. Marino 6 , A. Vallone 6 ,
N. Serrao 7 , V. Pisani 1 , C. Costa 1 , A. Terremoto 2 ,
G. Foti 3 , L. Cosco 4 , M. Calderazzo 5 ,
D. Corigliano 5 , P. Scordo 1 ,
C. Torti 1 , and the CalabrHIV Study Group
1 Unità Operativa di Malattie Infettive, Azienda
Ospedaliera Universitaria “Mater Domini”,
Università “Magna Graecia” Catanzaro, Italy
Ospedaliera Cosenza, Italy
Ospedaliera “Bianchi Melacrino Morelli” Reggio
Calabria, Italy
4 Unità Operativa di Malattie Infettive, Ospedale
“Pugliese” Catanzaro, Italy
5 Unità Operativa di Malattie Infettive, Presidio
Ospedaliero Lamezia Terme, Italy
“Jazzolino” Vibo Valentia, Italy
Crotone, Italy
Introduction and Aims HIV infected patients suffer from premature aging, putting them at risk of non-infectious co-morbidities
at younger ages than the general population. Aim of this study
is to evaluate for the first time prevalence of non-infectious
co-morbidities and of multi-morbidity in HIV/HCV co-infected
patients with respect to HIV mono-infected ones.
Materials and Methods The CalabrHIV observational cohort
includes all HIV patients followed by infectious disease centers
in the Calabria Region. Epidemiological, clinical and demographic characteristics were collected in a common database.
Non-infectious co-morbidities were recorded: cardiovascular diseases, hypertension, diabetes, renal failure and bone fractures.
Multi-morbidity was defined as ≥2 non infectious co-morbidities
occurring in the same patient.
Results 549 patients were selected (69% males, 37% >50 years).
Main risk factors were sexual promiscuities (50%) and IVDU (34%).
34% patients were HIV/HCV co-infected, 7% HBsAg carriers and
2% had triple co-infections. Hypertension was most frequent (25%
overall). Multi-morbidity was higher from 50 years of age (30%
vs. 6%; p < 0.0001). HIV co-infected patients had more frequently
AIDS (39% vs. 21%; p < 0.0001). A statistically significant difference in multi-morbidity percentage was observed in patients aged
>50 years with HCV and/or HBV co-infection than in HIV monoinfected ones (70% vs. 50%; p = 0.0037) (Figure 1). This effect was
not driven by the oldest subjects since an increasing significance
was found with increase in age starting from 40 years (39.5% vs.
e50
Figure 1. a) % Multi-morbidity in HIV mono infected patients; b)% multi-morbidity
in HIV patients co-infected with viral hepatitis.
26%; p = 0.0293) and, at >60 years of age, the difference was even
inverse (54% vs. 92%; p = 0.026).
Conclusions Even though co-infected patients are no longer
“special” with regard to HCV-treatment response, they are still
“fragile”. Prevention and early detection of non infectious comorbidities is important in these patients, especially in the young.
were detected with an intra-patient prevalence ranging from
0.11% for rtA181 T to 99.98% rtL180 M.
Analysing HBsAg a-determinant, 48.4% (30/62) of patients
carried ≥1 immune-escape mutation (intra-patient prevalence
range:0.2-100%). Vaccine-escape mutations were found in 11.4%
of patients, all genotype D-infected. This is the case of sG145R,
sM133L, and sP120S, detected with intra-patient prevalence ranging from 3.9% to 99.9% for sG145R, from 1.9% to 16.8% for sM133L,
and always of 100% for sP120S.
Stop-codons were found in 19.3% patients (intra-patient prevalence range:1.6-47.5%). They occurred at 11 HBsAg-positions
including also 172 and 182, known to increase HBV oncogenicpotential.
Finally, by Shannon-Entropy, specific HBsAg-positions correlated with the lack of HBsAg seroconversion in genotype-D. In
particular, positions 130 and 133 (localized in HLA-class-II epitope) were found mutated only in patients not developing anti-HBs
(Shannon-Entropy mean ± SE:1.98 ± 0.01 vs 0, and 1.95 ± 0.03 vs 0,
respectively, P < 0.05).
Conclusions: AHB is characterized by a complex coexistence
of viral-quasispecies, some of them with reduced antigenicity/immunogenicity, enhanced oncogenic-potential and altered
drug-susceptibility. These viral-variants may induce severe and/or
difficult-to-treat forms of HBV-infection (es. HBV-reactivation),
and might affect the efficacy of current HBV-vaccination strategy.
F-16
F-15
UPDATED CUT OFF VALUES OF LIVER STIFFNESS
TO MAXIMIZE TREATMENT OUTCOME OF
INTERFERON FREE ANTI HCV REGIMENS
A HIGH HBSAG GENETIC COMPLEXITY CAN
INFLUENCE HBV IMMUNOGENICITY IN THE
SETTING OF ACUTE INFECTION
A. Colli 1 , M. Fraquelli 2 , D. Prati 3 , A. Aghemo 4 ,
D. Conte 2 , M. Colombo 4 , G. Casazza 5
A. Battisti 1 , M. Aragri 1 , N. Coppola 2 , C. Alteri 1 ,
C. Sagnelli 3 , M. Pisaturo 2 , M.C. Bellocchi 1 ,
R. Salpini 1 , M. Starace 2 , D. Armenia 1 , L. Carioti 1 ,
M. Pollicita 1 , E. Sagnelli 2 , C.F. Perno 1 , V. Svicher 1
1
Department of Experimental Medicine and Surgery,
University of Rome “Tor Vergata”, Rome, Italy
Italy
Background. To characterize HBV RT and HBsAg geneticheterogeneity in acute HBV (AHB) infected patients and to define
their clinical value.
Methods. 62 HBsAg + and IgM/anti-HBc+ patients with clinical
and biochemical signs of AHB (44 genotype-D and 18 genotype-A)
were enrolled from 2000 to 2010. Plasma-samples obtained at firstobservation were analyzed by ultra-deep sequencing (UDPS) for
drug-resistance and immune-escape mutations. Shannon-Entropy,
weighted for intra-patient prevalence of each mutated-position,
was used to measure the extent of HBsAg amino-acid variability.
Results. 75.8% of patients were male with median(IQR)
age of 36(29-46) years. Median(IQR) ALT and HBV-DNA were
2,544(1,938-3,078)U/L and 5.9(4.5-7.4)log10 IU/ml. 61/62 (98.4%)
patients became HBsAg-negative with 33/61 (54.1%) developing
also anti-HBs.
By UDPS, 8.1% (5/62) of patients carried ≥1 drug-resistance
mutation (rtV173L/rtL180 M/rtA181 T/rtA194 T/rtM204I). They
1 Dipartimento di Medicina Interna, Ospedale A
Manzoni, Lecco, Italy
2 Unità di Gastroenterologia ed Endoscopia,
Fondazione IRCCS Ospedale Maggiore Policlinico,
Università degli Studi di Milano, Milano, Italy
3 Dipartimento di Medicina Trasfusionale ed
Ematologia, Ospedale A Manzoni, Lecco, Italy
4 Unità di Gastroenterologia ed Epatologia,
Fondazione IRCCS Ospedale Maggiore Policlinico,
Università degli Studi di Milano, Milano, Italy
5 Dipartimento di Scienze Biomediche e Cliniche,
Ospedale Luigi Sacco, Milano, Italy
BACKGROUND Interferon-free anti-HCV regimens will change
the paradigm of hepatitis C treatment.Yet due to money constrains
treatment access relies on pretreatment patients stratification for
disease severity using non-invasive tests including liver stiffness
measurement (LSM), whose cut-off values have been established
to maximize its diagnostic accuracy.Now cut offs are required to
maximize the clinical benefit of treatment. Based on the prevalence of different fibrosis stages and the harm/benefit ratio(H/B)
of different treatments, cut-off values can be obtained to drive the
treatment decision, i.e.to identify patients with the minimum level
of hepatitis requiring treatment.
METHODS 11 hepatologists in different tertiary centers
quantified the expected H/B, based on published data and clinical judgment, for different scenarios: two different disease
stages (F≥2orF≥3) for threetreatment regimens (PEGIFN + RBV
or PEGIFN + RBV + 1ST generation PI or IFNfree regimens). From a
cohort of 750 consecutive HCV patients we obtained two ROC
e51
CONCLUSIONS The estimated H/B of interferon-free regimens is
extremely low and thus the LSM cut-off values to initiate treatment
are lower than the ones predicting maximal diagnostic accuracy,
either assuming to treat patients with F≥3 or all comers with F≥2.
1)Cantor et al. J Clin Epidemiol 1999
Methods: 584 patients undergoing surgery with curative intent
for ICC between 1990 and 2013 at one of 12 participating institutions were identified. A nonmixture cure model was adopted to
compare mortality after hepatic resection to the mortality expected
for the general population matched by sex and age.
Results: Median, 1-, 3-, 5- years disease-free survival was 10
months, 44%, 18% and 11%; the corresponding overall survival was
27 months, 75%, 37% and 22%. The probability of being cured from
ICC was 9.7% (95% confidence interval 6.1-13.4%). The mortality of
patients undergoing surgery for ICC was higher than the general
population until year 10, when the patients alive without tumor
recurrence can be considered cured with 99% certainty. Multivariate analysis showed that cure probabilities range from 25.8% (time
to cure 9.8 years) in patients with single ICC ≤ 5 cm, and without
minor vascular invasion, poor grade, lymph nodes metastases, and
periductal histology, to < 0.1% (time to cure 12.6 years) in patients
with all these six risk factors.
Conclusions. The cure model indicates that statistical cure is
possible in patients undergoing hepatic resection for ICC. A model
to calculate the cure fraction and time to cure in each clinical scenario was provided (http://www.livercancer.eu/CCC cure model).
F-17
F-18
CAN HEPATIC RESECTION PROVIDE A
LONG-TERM CURE TO PATIENTS WITH
INTRAHEPATIC CHOLANGIOCARCINOMA?
AUTOFLUORESCENCE DETECTION OF LIVER
OXIDATIVE DAMAGE PRODUCTS
curves assessing LSM accuracy in diagnosing F≥2 or F≥3 (RS:liver
biopsy). The prevalence,p(D), of F ≥2 or F ≥3 (D) was respectively
53% and 30%. For each scenarios the optimal cut-off value maximizing the expected utility of LSM was identified on the ROC curve
as the point with slope equal to equation(1). Normal median LSM
values in a control population (#1001 #participants) from the same
area was 4.7 kPa (2.6-7.6, percentile 5th - 95th).
RESULTS Mean H/B value for the three different therapeutic
regimens according to two different liver disease scenarios:
Mean H/B
LSM(kPa)
cut-offs
PEGINF +
RBV
F≥2
F≥3
PEGINF +
RBV + PI
F≥2
F≥3
IFN-free
F≥2
F≥3
1/3.1
6.1
1/3.7
9.4
1/2.8
6.4
1/4.1
9.3
1/8.3
4.1
1/10
7.1
G. Spolverato 1,∗ , A. Vitale 2,∗ , A. Cucchetti 3 ,
S. Alexandrescu 4 , H.P. Marques 5 , L. Aldrighetti 6 ,
T.C. Gamblin 7 , S.K. Maithel 8 , C. Pulitano 9 ,
T.W. Bauer 10 , F. Shen 11 , G.A. Poultsides 12 ,
J.W. Marsh 13 , T.M. Pawlik 1
1 The Johns Hopkins University School of Medicine,
Baltimore, MD, United States of America
2 Unità di Chirurgia Epatobiliare e Trapianto Epatico,
Azienda Ospedaliera-Università di Padova, Padova,
Italy
3 Department of Medical and Surgical Sciences, S.
Orsola-Malpighi Hospital, Alma Mater Studiorum University of Bologna, Bologna, Italy
4 Fundeni Clinical Institute, Bucharest, Romania
5 Curry Cabral Hospital, Lisbon, Portugal
6 Ospedale San Raffaele, Milan, Italy
7 Medical College of Wisconsin, Milwaukee, WI,
United States of America
8 Emory University, Atlanta, GA, United States of
America
9 University of Sydney, Sydney, Australia
10 University of Virginia, Charlottesville, VA, United
States of America
11 Eastern Hepatobiliary Surgery Hospital, Shanghai,
China
12 Stanford University, Stanford, CA, United States of
America
13 University of Pittsburgh Medical Center,
Pittsburgh, PA, United States of America
Background: A patient can be considered statistically cured
from the specific disease when the mortality returns to the same
level as that of the general population. Cure models have never been
applied to patients undergoing hepatic resection for intrahepatic
cholangiocarcinoma (ICC). We sought to assess the probability of
being statistically cured from ICC by hepatic resection.
A.C. Croce 1,2 , A. Ferrigno 3 , V.M. Piccolini 1 ,
L.G. Di Pasqua 3 , C. Berado 3 , G. Bottiroli 1,2 ,
M. Vairetti 3
1
IGM-CNR, Pavia, Italy
Department of Biology & Biotechnology, University
Pavia, Italy
3 Department of Internal Medicine and Therapy,
University of Pavia, Pavia, Italy
2
Introduction- Oxidative stress in fatty livers enhances the risk
of disease progression or organ dysfunction in surgery and transplantation. An experimental model of induced-NASH showed a
liver metabolism subversion, reflected by tissue autofluorescence
(AF) alterations. These consisted in changes in NAD(P)H/flavin AF
related contributions -ascribable to redox metabolism alterationsand a rising in vitamin A, protein, and lipofuscin-like-lipopigment
emission signals.
Aim- Lipofuscin-like-lipopigments derive from the oxidation of
unsaturated lipid. Therefore fluorescing fatty acids and oxidized
products were investigated as early oxidative biomarkers in livers
with mild lipid accumulation, under different preservation conditions.
Materials and Methods- Rat livers(controls and 2 week
administered methionine/choline-deficient diet-MCD) were isolated and submitted to 6-h Cold Storage (CS) or subnormotermic
Machine Perfusion (MP), followed by reperfusion (37 ◦ C, oxygenated medium). AF spectra recorded in vivo (exc 366 nm), via
fiber-optic probe, were analyzed by curve fitting procedure, to
estimate each endogenous fluorophore contribution to the overall
emission, similarly to in situ biochemical analysis. Tissue oxidative stress (TBARS, and GSH) and mitochondrial dysfunction
(ATP/ADP) were assayed using conventional methods. Results. Both
CS or MP preservation affected liver AF properties. The response
of NAD(P)H/flavin redox state to oxygenation and temperature
was consistent with MCD induced mitochondrial dysfunction.
Lipofuscin-like-lipopigment AF signals rose in both controls and
MCD livers, in a good correlation with TBARS. Independent from
e52
preservation procedure, the phenomenon was significantly greater
in MCD than in control livers, indicating a strong influence of fatty
liver oxidative stress. Fluorescing fatty acids were not depleted,
consistently with dynamic equilibria in liver lipid pool composition.
Conclusions. The real time AF analysis allows to monitor oxidative effects even in a model of mild metabolic alterations, providing
an AF support in experimental investigations on drug response and
toxicity, and in the set-up of innovative organ preservation strategies.(Supported by Fondazione Cariplo, grant n◦ 2011-0439).
F-19
PREVALENCE AND RISK FACTORS OF METABOLIC
SYNDROME AFTER LIVER TRANSPLANTATION: A
SINGLE CENTRE EXPERIENCE
V. Pepe 1 , G. Germani 1 , A. Ferrarese 1 , A. Zanetto 1 ,
I. Bortoluzzi 1 , E. Nadal 1 , F.P. Russo 1 ,
M. Senzolo 1 , E. Gringeri 2 , U. Cillo 2 , P. Burra 1
1
Multivisceral Transplant Unit, Department of
2 Hepatobiliary Surgery and Liver Transplant Unit,
Padua University Hospital, Padua, Italy
Background and Aim: Metabolic syndrome (MS) is common
after liver transplant (LT), increasing long term mortality and morbidity. The aim of this study was to evaluate short and long-term
prevalence and risk factors of MS after LT.
Materials and methods: patients who underwent LT at Padova
Liver Transplant Center between January 2000 and March 2013
(retrospective cohort) and between April 2013 and April 2014
(prospective cohort) and followed-up at Multivisceral Transplant
Unit were included. Patients < 18 years, who underwent multiorgan transplantation or re-LT and with pre-LT MS were excluded.
For each patient general and metabolic variables and donor characteristics were recorded.
Results: In the retrospective cohort 161 patients (120 M) were
included. The most common indication to LT was HCV-related
cirrhosis (49.1%). A post-LT significant increase in BMI values, in
diabetes and hypertension prevalence and in total cholesterol and
triglyceride levels was found compared to pre-LT values. At a mean
post-LT follow-up of 6.9 ± 4.2 years 81/161 (50.3%) patients developed MS. Recipient male sex (OR 2.36, p = 0.045), a higher pre-LT
BMI (OR per unit 1.14, p = 0.03), and the presence of pre-LT diabetes
(OR 5.98, p = 0.04) were associated with the development of postLT MS. In the prospective cohort 15 patients were included (10 M).
At 3, 6 and 12 months after LT a significant increase in BMI values, diabetes and hypertension prevalence and in cholesterol and
triglyceride levels was found compared to pre-transplant values
and 5/15 (33.3%), 3/11 (27.3%) e 4/10 (40%) patients developed MS.
Conclusions: post-LT MS affects nearly half of LT recipients,
starting early after LT. Recipient male gender, pre-LT diabetes and
increased BMI are risk factors for MS after LT. Lifestyle modifications, should be recommended to transplanted recipients starting
in the early post-LT period, thus preventing body weight gain and
the associated abnormalities and reducing incidence of post-LT MS
and the related cardio-vascular events.
F-20
SHORT-TERM OUTCOME POST LIVER
TRANSPLANTATION OF CIRRHOTIC LISTED
OUTPATIENTS WITH FLUCTUATIONS OF EGFR
F. Fiacco 1 , F. Tinti 1 , I. Umbro 1 , A. Zavatto 1 ,
S. Ginanni Corradini 2 , M. Rossi 3 , P.B. Berloco 3 ,
A.P. Mitterhofer 1
1 Department of Clinical Medicine, Nephrology and
Dialysis Unit, Sapienza University of Rome, Rome,
Italy
2 Department of Clinical Medicine, Gastroenterology
Unit, Sapienza University of Rome, Rome, Italy
3 Department of General Surgery, Organ Transplant
Unit “Paride Stefanini”, Sapienza University of Rome,
Rome, Italy
Introduction-In cirrhotic patients listed for liver transplantation (ESLD-wLT) renal dysfunction results from systemic conditions
that affect both liver and kidney leading to kidney ischemia. These
patients are prone to estimated-glomerular filtration rate (eGFR)
changes related to pre-renal conditions. Nephrologic guidelines
consider eGFR modifications common and not necessarily indicative of progression to chronic kidney disease (CKD), but their
meaning in ESLD-wLT outpatients is not clear and assessment of
eGFR is challenging.
Aim-to evaluate eGFR variations in ESLD-wLT outpatients and
outcome post-LT related to short-term renal, graft and patient survival.
Methods-Single centre retrospective study of 51 ESLD-wLT outpatients. All available GFR values estimated by MDRD formula were
collected considering eGFR at listing (eGFR-list), eGFR at transplant
(eGFR-LT), the lowest (eGFR-min) and the highest value of eGFR,
further defined as baseline (eGFR-baseline). Fluctuations (FeGFR)
were considered as percentage variation between eGFR-baseline
and eGFR-min. Fluctuations of eGFR > 50% were defined as drop.
Acute Kidney Injury (AKI) and CKD were defined according to
KDIGO-Guidelines. Short-term graft function, AKI post-LT and 30days survival after-LT were assessed.
Results-All patients presented FeGFR. Estimated-GFR-LT was
not different from eGFR-list. Estimated-GFR drops were observed
in 18/51 patients (DeGFR + group). These patients presented higher
prevalence of pre-LT CKD, higher MELD score and bilirubin both at
listing and LT and lower sCr-min. AKI post-LT stages 2-3 occurred
more frequently in DeGFR+ group compared to DeGFR- group (8/18
vs 3/33,p0.01). Patients in DeGFR+ group required more plasma
infusions and inotropic support during LT, while no differences
were detected in early graft function and survival after-LT.
Conclusions-Estimated-GFR fluctuations are common in ESLDwLT outpatients and are not indicative of worsening of renal
function from listing to LT. Estimated-GFR drops are more frequent
in patients with CKD and worse condition of cirrhosis and are associated with AKI post-LT. Monitoring pre-LT eGFR may be useful in
predicting post-LT outcome.
F-21
F-22
FIRST STEPS TOWARDS UNDERSTANDING THE
DYNAMIC EVOLUTION OF GUT MICROBIOTA IN
DIFFERENT STAGES OF LIVER DISEASE
PREVALENCE OF HEPATITIS E IN TERTIARY
HOSPITAL CENTRE IN NORTHERN ITALY
F.R. Ponziani 1 , S. Pecere 1 , F. Paroni Sterbini 2 ,
V. Petito 1 , M. Siciliano 1 , T. Di Rienzo 1 ,
A. Palladini 2 , D. Zambrano 1 , F. Franceschi 1 ,
E. Gaetani 1 , F. Scaldaferri 1 , L. Masucci 2 ,
M. Sanguinetti 2 , A. Gasbarrini 1
1 Internal Medicine Department, Gastroenterology
Division, Catholic University Of Sacred Heart of
Rome, Rome, Italy
2 Microbiology Department, Catholic University Of
Sacred Heart of Rome, Rome, Italy
INTRODUCTION: Liver cirrhosis is associated to intestinal barrier derangement and dysbiosis. AIM: To preliminary describe gut
microbiota modifications in a population of Italian patients with
liver cirrhosis.
PATIENTS AND METHODS: Fecal samples were collected in
13 cirrhotic patients and 10 healthy controls, no exposed for
at least one month to antibiotics, prebiotics, probiotics and
bowel colonoscopy preparation. Gut microbiota composition was
assessed by a metagenomic gene-targeted approach (16S rRNA),
following DNA isolation from stool samples stored at–80 ◦ C. Data
were analyzed in Qiime. Non parametric tests were used for the
final statistical analysis.
RESULTS: The enterotypification of the 23 subjects was comparable to that of the Western general population, with a prevalence of
Bacteroides (enterotype 1, 77%, often overlapping with enterotype
3, 18%) with no inter- or intra-group differences between cirrhotics
and healthy controls. A trend toward an unbalance between the Firmicutes to Bacteroidetes ratio was found throughout the different
Child Pugh stages (close to 1 in Child A, reduced to half in Child B
and to ¼ in Child C p = 0.081). Compared to healthy individuals, cirrhotics microbiota was more enriched in Bacilli (Lactobacillaceae,
Streptococcaceae, Enterococcaceae; p = 0.001), especially in case of
intermediate stage liver disease (abundance in Child B patients
48.6%; p = 0.008). Clostridia (Clostridiaceae, Ruminococcaceae,
Veillonellaceae, Lachnospiraceae, Clostridiaceae, Peptostreptococcaceae, Peptococcaceae) were the most abundant in the initial
phases of the disease (62.3% in Child A patients p = 0.06). Patients
with at least one clinical sign of decompensation lost the relative abundance of Clostridia (abundance 62.3% in compensated
patients vs 20.8% in decompensated ones vs 35.4% in healthy controls p = 0.04).
CONCLUSION: Liver cirrhosis is a fascinating model of gut microbiota modifications, that are connected with the progression of liver
impairment.
e53
V. Zuccaro 1 , P. Columpsi 1 , S. Paolucci 2 ,
M. Mariani 1 , S. Toppino 1 , A. Malfitano 1 ,
A. Parisi 1 , S.F.A. Patruno 1 , F. Baldanti 2 , R. Bruno 1
1 Department of Infectious Diseases–IRCCS San
Matteo - University of Pavia, Pavia, Italy
2 Virology Unit IRCCS San Matteo - University of
Pavia, Pavia, Italy
Introduction: In developed countries, hepatitis E is a zoonosis
usually caused by the genotypes 3 and 4. In Italy the prevalence of
anti-HEV is 1-5% among the healthy population. A large prospective
study has shown that HEV accounts for approximately 10% of acute
non A, non B hepatitis cases in Italy.
Aim: The aim of the study was to evaluate the prevalence of hepatitis E among patients admitted to the Division of Infectious and
tropical Disease for acute hepatitis without evidence of autoimmunity and antibody against HAV-HBV-HCV.
Materials and Methods Results: A total of 970 patients were
admitted to Division of Tropical and Infectious Diseases from 2013
to 2014. Among them 27 had acute hepatitis and 3 (11%) were positive for hepatitis E. All were anti-HEV IgM and IgG positive and
were also positive for HEV-RNA, viral genotype was 3 subtype F
in all cases. All HEV patients were living from Pavia and denied
having travelled abroad. Diagnosis of hepatitis E was based on the
presence of IgM anti-HEV and confirmed by the detection of HEVRNA - RTPCR on patients samples. Viral isolates were sequenced
to characterize subtypes. Two cases with acute disease had a selflimited course with AST-ALT normalization within 3-6 weeks. One
patient died from acute on chronic liver disease. All cases showed
high level of amylases during the acute phase.
Conclusions: This is the first report of hepatitis E in the
Pavia area and it confirms the increasing trend of prevalence of
autochthonous hepatitis E in our country. The increased prevalence
might be related either to the spread of the HEV virus in our country and to the implementation of HEV screening in patients with
acute hepatitis. In the case of acute hepatitis with high amylases
level should prompt exclude HEV infection.
F-23
IMPACT OF RIFAXIMIN IN THE PREVENTION OF
BACTERIAL INFECTIONS IN CIRRHOSIS
M. Mariani 1 , L. Scudeller 2 , S.F.A. Patruno 1 ,
V. Zuccaro 1 , P. Columpsi 1 , S. Toppino 1 ,
A. Malfitano 1 , A. Parisi 1 , G. Filice 1 , R. Bruno 1
1
Division of Infectious and Tropical Diseases–IRCCS
San Matteo - University of Pavia, Pavia, Italy
2 Scientific Direction, IRCCS Policlinico San Matteo,
Pavia, Italy
Background and aims: Infectious events represent a leading
factor able to accelerate the progression from compensated to
decompensated stage of cirrhosis, with a subsequent worsening
of the prognosis. Due to the high frequency of infectious complications in cirrhotic patients, a great effort was made in order to
improve the survival rate. Rifaximin was advanced as a possible
primary prophylactic therapy against spontaneous bacterial infections. We retrospectively investigated the role of this antibiotic in
e54
reducing the proportion of infected patients in treated and nontreated groups.
Methods: We enrolled 649 patients whose clinical and personal
data, prescribed therapy, microbiological findings and laboratory
tests were collected from previous discharge letters and our institution database. We excluded patients with ACLF and acute hepatitis.
Aetiology, drug resistance and site of infection were collected as
well.
Results: Risk of developing bacterial infections was significantly
lower in patients treated with rifaximin (6 month of treatment, 10
days per month, 400gr t.i.d.) (OR 0.29; 95% CI 0.20-0.40, p < 0.001)
compared to those not treated. Moreover, in our cohort we find
out a prevalence of infections sustained by Gram-positive bacteria
(61.5% of isolations).
Conclusions: Rifaximin continuous treatment turned out to be
a very effective factor involved in bacterial infections prevention
in cirrhotic patients. Our study showed a shift in general bacterial
prevalence among pathogens to Gram positive, this finding should
be considered as well before administering an empirical therapy in
cirrhotic patients.
F-24
SERUM ALPHA-FETOPROTEIN MODIFICATION
(DELTA-AFP) IS A HIGHLY SPECIFIC TOOL TO
SUSPECT HCC RECURRENCE AFTER LIVER
TRANSPLANTATION
F.R. Ponziani 1,∗ , S. Bhoori 1 , M. Bongini 1 ,
M. Flores 1 , C. Muscarà 1 , V. Mazzaferro 1
1 Liver Transplant, Hepatobiliary and
Gastrointestinal Surgery, Hepatology National
Cancer Insitute of Milan, Milan, Italy
Introduction: Alpha-fetoprotein (AFP) is the main tumor
marker of hepatocellular carcinoma (HCC). Despite AFP values at
listing have been used to predict HCC recurrence after liver transplantation (LT), its role in patients’ surveillance after LT has not
been established yet.
Aim: To investigate the usefulness of serum AFP modification
to suspect recurrence in patients who previously underwent LT for
HCC.
Materials and Methods Results: We investigated all consecutive patients undergoing LT for HCC within Milan or up-to-seven
criteria from January 2004 to December 2013. Serum AFP was
collected at the time of LT and every 6-months for 2 years and
every year thereafter. Patients surveillance consisted in semiannual abdominal ultrasound or CT scan for the first 5-years, then
every year.
231 LT recipients were investigated (median age 61 (14-64), 89%
males, 30% HBV, 51.5% HCV, 4% coinfected, 10% alcohol, 8% other,
4% HIV); 31 of them (13%) experienced HCC recurrence. Among
HCC recurrence-free patients, 24 (10%) developed cirrhosis or liver
failure, severe hepatitis or alcohol abuse during follow-up; since
their median AFP values had higher peaks, they were excluded to
avoid biases. Univariate analysis failed to demonstrate any association between serum AFP at any timepoint and HCC recurrence,
while its modification (difference between the highest AFP value
detected during follow-up or the value detected at recurrence,
and the lowest serum AFP value, named “delta-AFP”) was strongly
associated with HCC recurrence (p < 0.0001). Multivariable analysis confirmed delta-AFP as the only independent predictive factor
of HCC recurrence (OR 1.28, 95%CI 1.116, 1.470; p = 0.001). The
accuracy of delta-AFP in predicting recurrence was 71.5% (95%CI
65% > 77.5%), with the best identified cut-off value of 6.57 ng/ml
(sensitivity 61.3% specificity 100%, NPV 96% PPV 100%).
Conclusions: Delta-AFP is a very easy-to-use and highly specific
tool to suspect HCC recurrence after LT.
F-25
CLINICAL PRESENTATION, TREATMENT AND
OUTCOME OF ALCOHOL- AND HEPATITIS C
VIRUS- RELATED HEPATOCELLULAR
CARCINOMA IN THE NEW CENTURY:
COMPARISON BEFORE AND AFTER ADJUSTMENT
WITH PROPENSITY SCORE ANALYSIS
L. Bucci 1 , F. Garuti 1 , V. Camelli 1 , B. Lenzi 1 ,
M. Bernardi 1 ,
F. Trevisani 1 , for the Italian Liver Cancer (ITA.LI.CA) group
1 Dipartimento di Scienze Mediche e Chirurgiche Alma Mate Studiorum - University of Bologna,
Bologna, Italy
Introduction and aims: Hepatitis C virus (HCV) and alcohol
abuse are the main causes of hepatocellular carcinoma (HCC) in
Western world, accounting for about 60% and 20% of cases, respectively. This study aimed at comparing the clinical presentation and
outcome of alcohol- and HCV- related HCC diagnosed in the new
century.
Materials and Methods: 1844 HCV and 636 alcoholic patients
from the Italian Liver Cancer (ITA.LI.CA) database, diagnosed with
HCC from January 2000 to December 2012 were compared for age,
gender, type of diagnosis, Barcelona Clinic Liver Cancer (BCLC),
tumor gross pathology, portal vein thrombosis (PVT), esophageal
varices, model for end stage liver disease (MELD), Child-Pugh class,
alpha-fetoprotein (AFP), treatment and survival. For survival analysis, a propensity matching model was performed to minimize
imbalances of prognostic factors between groups.
Results: Underlying cirrhosis was present in 96% of both
alcoholic and HCV patients. Alcoholic patients were significantly
younger and more likely male, diagnosed with a tumor outside
surveillance, in intermediate and terminal BCLC stage and in a
worse hepatic function setting. Consequently, the overall survival
was decreased in alcoholic compared to HCV patients [median
(95% C.I.): 34.5 (28.7-40.2) vs 43.6 (40.9-46.3) months; p < 0.001]. A
one-to-one matching after propensity score adjustment according
to age, gender, type of diagnosis, BCLC stage, tumor gross pathology, PVT and treatment was performed. This analysis selected 354
patients per group, well balanced for baseline prognostic factors.
The overall survival did not differ any longer between alcoholic and
HCV group [median (95% C.I.): 42.6 (36.5-48.8) vs 38.6 (31.2-45.9)
months; p = 0.756].
Conclusions: Alcoholic patients present with a more compromised liver function and a more advanced HCC, less frequently
detected during surveillance. Nevertheless, alcoholic etiology per
se does not affect survival as compared with HCV-related cases,
since it was similar in the two groups once adjusted for the confounding factors.
F-26
F-27
ENOS DYSFUNCTION IN STEATOSIS AND
STEATOHEPATITIS
SOFOSBUVIR PLUS RIBAVIRIN IN HCV-INFECTED
CIRRHOTIC PATIENTS AWAITING LIVER
TRANSPLANTATION: PRELIMINARY DATA FROM
A SINGLE-CENTRE EXPERIENCE
M. Masarone 1 , A. Carrizzo 2 , A. Federico 3 ,
V. Rosato 4 , E. Claar 5 , C. Vecchione 6 , M. Persico 1
1
Internal Medicine and Hepatology Unit, University
of Salerno, Salerno, Italy
2 Vascular Physiopathology Unit IRCCS, INM
Neuromed, Pozzilli (IS), Italy
3 Gastroenterology Division - Second University of
Naples, Italy
4 Internal Medicine and Hepatology Department,
Second University of Naples, Italy
5 Internal Medicine Department, Hepatology Unit,
Ospedale Evangelico Villa Betania, Naples, Italy
6 Department of Medicine and Surgery, University of
Salerno, Salerno, Italy
Background: Non-Alcoholic-Fatty-Liver (NAFLD/NASH) has
been associated to cardiometabolic syndrome that is a risk
factor for cardiovascular diseases (CVD). The basis of CVD is
oxidative stress. Endothelial Nitric-Oxide-Synthase (eNOS) dysfunction has been widely involved in oxidative stress and CVD.
Although NAFLD/NASH and CVD are associated to the same factors, NAFLD/NASH represents itself an independent CVD risk-factor.
Even diabetics show higher rates of CVD if they have NASH. eNOS
derangement has been demonstrated in animal experimental models of NAFLD/NASH. Although clinical and “sublinical” markers
(i.e.“intima-media-thickness”) seem to have confirmed this suspicion, nevertheless, no experimental studies on humans have
directly demonstrated that eNOS dysfunction is associated with
NAFLD/NASH.
Aim: to directly demonstrate eNOS derangement in
NAFLD/NASH.
Patients and methods: 18 patients (13 M,5F), coming in our
department of Internal Medicine for NAFLD/NASH, were consecutively enrolled. Every patient underwent a clinical evaluation and
a liver biopsy after informed consent. Patients were divided in two
groups according to the presence of NAFLD or NASH. We measured
eNOS function by evaluating the vasorelaxation activity induced on
isolated-mice-vessels by platelet-rich-plasma of peripheral blood,
and by immunoblot-assays for platelet-derived eNOS (p-eNOS).
Collected data were compared to those from an age-, sex-matched
group of healthy volunteers from a local blood bank. All subjects
were non-smokers and had no active CVD.
Results:Of the 18 pts, 7 (38,8%) had NAFLD and 11 (61,7%)
had NASH at the liver biopsy. No statistically significant differences were found between the two groups and controls for age,
sex, BMI, ALT, hypertension, diabetes, dyslipidaemia, obesity and
metabolic syndrome. Vascular reactivity curves demonstrated a
reduced activity of eNOS in patients with NAFLD/NASH in respect
to controls(p < 0.005). Moreover, immunoblot-assays for p-eNOS
demonstrated a significantly lower expression in NAFLD/NASH
patients in respect to controls (p < 0.007).
Conclusions: we directly demonstrated that eNOS function is
reduced in NAFLD/NASH. Endothelial dysfunction may be considered as one of the pathophysiological mechanisms of liver damage
in NAFLD/NASH.
e55
S. Martini 1 , S. Strona 1 , D. Arese 1 , M. Sacco 1 ,
R. Romagnoli 2 , F. Tandoi 2 , A. Ottobrelli 1 ,
M.R. Torrani-Cerenzia 1 , F. Balzola 1 , M. Salizzoni 2 ,
M. Rizzetto 1
1 Gastrohepatology Unit, Department of Medical
Sciences, A.O.U. Città della Salute e della Scienza di
2 Liver Transplant Centre, General Surgery Unit,
Department of Surgical Sciences, A.O.U. Città della
Salute e della Scienza di Torino, Turin, Italy
Background and aims: Clearance of HCV infection before liver
transplantation (LT) prevents recurrence after LT. We evaluated
safety and efficacy of antiviral therapy (Tx) with sofosbuvir (SOF)
plus ribavirin (RBV) in HCV-infected cirrhotic patients (pts) before
LT.
Methods: Since June 2014 to date we enrolled 24 HCV cirrhotic
pts with HCC within Milan criteria (15 pts) and/or decompensated
liver disease. Median MELD 13 (range 7-19); Child-Pugh score B
71% and C 17%; 83% male, mean age 55.4 years, mean BMI 25.8.
Mean baseline HCVRNA 6.17 log10 IU/mL (4.91- 7.11 log10 IU/mL);
GT1b 55%, GT1a 4%, GT2 4%, GT3 29%, GT4 8%; IL28CC 20%; 63%
experienced. Planned treatment: SOF 400 mg/die (compassionate
use) and RBV (weight-based) for 48 weeks or until LT.
Results: Antiviral treatment resulted in rapid suppression of circulating virus (median decrease HCVRNA: 3.27 log10 IU/mL after 1
week of Tx); 10/20 pts (50%) at week 4, 14/14 week 8, 10/10 week 12
and 7/7 week 16 on Tx were HCVRNA negative. No serious adverse
events were observed. RBV reduction and/or epoetin administration was required in 25% of pts and 3 were hospitalized due to
complications of liver disease. Of the 7 pts transplanted until now,
4 had HCVRNA < 15 IU/mL for a median of 62 days (31-91) and discontinued antiviral Tx at LT. All those 4 pts are currently HCVRNA
negative after a median follow-up post-LT of 45 days (16-69). 2 pts
underwent LT while still being HCVRNA positive after less than 4
weeks of Tx, and the last one was transplanted 3 days ago having
HCVRNA <15 IU/mL since 5 days.
Conclusion. Tx with SOF plus RBV induces rapid HCV clearance
and is well tolerated in cirrhotic pts awaiting LT. Preliminary postLT SVR4 is promising in pts undergoing LT after at least 1 month
from viral clearance.
e56
F-28
F-29
DISTRIBUTION OF STEM CELLS AND CANCER
STEM CELLS MARKERS IN LIVER PATHOLOGIES
AND THEIR INDICATION TO THE RESPONSE OF
THERAPY
LIVER TRANSPLANTATION (OLT) AND
CARDIOVASCULAR RISK: ASSESSMENT OF EARLY
ATHEROSCLEROTIC DAMAGE AND METABOLIC
DISORDERS IN PATIENTS UNDERGOING LIVER
TRANSPLANTATION
C. H.C. Sukowati 1,2 , B. Anfuso 1 , D. Pascut 1 ,
R. Patti 1,2 , N. Mezzina 1,2 , P. Tarchi 3 ,
S.L. Crocè 1,2 , C. Tiribelli 1,2
1
Fondazione Italiana Fegato, AREA Science Park
Basovizza, Trieste, Italy
2 Department of Medicine, Surgery, and Health
Sciences, University of Trieste, Trieste, Italy
3 Department of General Surgery, Teaching Hospital
Cattinara, Trieste, Italy
Introduction Recent studies had suggested different markers
to identify the population of stem cells (SC) and cancer stem cells
(CSC) in hepatocellular carcinoma (HCC). However, a study on the
distribution of the markers that may indicate disease progression,
nature of cancer, and response to therapy will be crucial in both
basic and translational research.
Aim To study the variation and distribution of various SC
and CSC markers during hepatocarcinogenesis and the association
between circulating CSC and HCC therapy.
Methods We analyzed 119 samples (12 normal tissues, 40 liver
cirrhosis (LC), 33 HCC; 34 blood samples of HCC). The comparison
of SC and CSC markers CD90, CD44, CD133, EpCAM, CD13, CK19,
CK7, and ABCG2 in each pathology groups was assessed by RTqPCR using MIQE guidelines. The effect of different treatments as
resection, radiofrequency, and chemoembolization to circulating
CSC was assessed by comparing positivity and expression on pretreatment (T0) with 1 month post-treatment (T1).
Results The expression of CD90 was positively correlated with
hepatocarcinogenesis moving from normal, LC, to HCC (p < 0.001)
confirmed by Western Blot; an increase in CD44 expression was
also observed. In contrast, the expressions of EpCAM, CD133, CK19,
and CK7 were increased only in LC. The CD13 expression was related
with HCV and MELD score (p < 0.05) while the ratio HCC:LC of
ABCG2 was associated with Edmonson-Steiner grade.
At T0, 80% of blood were positive for CSC CD90 and only 40% for
both CD133 and EpCAM. At T1, HCC resection reduced the expressions of CSC markers CD90, CD133, and EpCAM, radiofrequency
reduced CD133, while chemoembolization increased EpCAM.
Conclusion This study demonstrated that CSC CD90 marker is
increased during hepatocarcinogenesis. The detection of circulating
CSC may be used to study the response to therapy without taking
biopsy, it will be important in the diagnostic and prognostic value.
G. Pisano 1 , M. Orlandi 1 , F. Donato 2 , S. Zannoni 1 ,
C. Bertelli 1 , M.Porzio 1 , M. Colombo 2 , S. Fargion 1 ,
A.L. Fracanzani 1
1 Department of Pathophysiology and
Transplantation, Maggiore Policlinico Hospital
Foundation IRCCS, University of Milan, Milan, Italy
2 Division of Gastroenterology, Maggiore Policlinico
Hospital Foundation IRCCS, University of Milan,
Milan, Italy
BACKGROUND: Improved survival after orthotropic liver transplantation (OLT) has allowed to detect long-term consequences of
liver transplantation. Nowadays, cardiovascular and cerebrovascular diseases are the leading causes of morbidity and death among
transplant recipients. Metabolic syndrome and its individual components, (diabetes mellitus, hypertension, dyslipidemia, obesity)
contributing to cardiovascular complications are increasingly being
identified in these patients.
AIMS: To evaluate the prevalence of metabolic syndrome and of
subclinical atherosclerosis in patients with OLT at 6 month and at
1 year follow-up.
METHODS: We evaluated 42 patients: 27 on transplant list
(group 1) and 15 patients transplanted within 6 months (group
2). In all patients anthropometric, clinical and metabolic parameters were evaluated before transplant. We evaluated 1) carotid
stiffness (cPWV), intima-media thickness (cIMT), and presence of
plaques (CP), 2) echocardiographic parameters (E/A, thickness of
the interventricular septum, left ventricular mass) and 3) visceral
adiposity by epicardial fat thickness before and at 6 and 12 month
after transplant.
RESULTS: All metabolic parameters (Cholesterol, triglycerides,
fasting glucose), and blood pressure increased after OLT. Metabolic
syndrome were present in 19%, 57% and 54% of patients at time
0, 6 and 12 months respectively (p = 0.04), while the prevalence of diabetes did not significantly differ before and after OLT.
A significant increase of carotid stiffness (7.2 ± 1,5 vs 8.3 ± 1.6,
p = 0,05), and interventricular septum thickness (10.2 ± 0.4 vs
12.2 ± 2.2, p = 0,02) and decrease of diastolic function (E/A 1.2 ± 0.4
vs 0.86 ± 0.3, p < 0,01) was observed after 6 months from transplant. The cIMT and epicardial fat thickness significantly increased
after 12 months (0.76 ± 0.2 vs 0.89 ± 0.2, p = 0.05) and (5.7 ± 3.2 vs
8.0 ± 3.6, p = 0.05).
CONCLUSIONS: Metabolic Syndrome is highly prevalent in liver
transplanted patients and cardiovascular alterations occur very
early after liver transplantation. Recognition and promptly treatment of these conditions may impact long- term survival of these
patients.
F-30
F-31
SCREENING FOR THE IDENTIFICATION OF
AUTOIMMUNE OR LYMPHOPROLIFERATIVE
ONSET IN PATIENTS NAïVE TO HCV ANTIVIRAL
TREATMENT
THE FIRST STEP FOR THE PREVENTION OF
ASCITES IN PATIENTS WITH CIRRHOSIS: THE
IDENTIFICATION OF AN EASILY ACCESSIBLE
PREDICTOR OF THE FIRST ONSET OF ASCITES
F. Gulli 1 , U. Basile 2 , L. Colacicco 2 , L. Miele 1 ,
N. De Matthaeis 1 , P. Cattani 3 , G.L. Rapaccini 1
S. Piano 1,2 , M. Tonon 1,2 , S. Fasolato 1,2 ,
G. Bombonato 1 , A. Romano 2 , E. Gola 2 ,
A. Brocca 2 , F. Morando 2 , M. Cavallin 2 ,
P. Angeli 1,2
1 Department of Internal Medicine, Istitute of
Internal Medicine and Geriatry- Catholic University
of the Sacred Heart, Rome, Italy
2 Department of Laboratory Medicine, Institute of
Biochemistry -Catholic University of the Sacred
Heart, Rome, Italy
3 Department of Laboratory Medicine, Institute of
Microbiology -Catholic University of the Sacred
Heart, Rome, Italy
Introduction. Hepatitis C virus(HCV) may be responsible of
extra-hepatic manifestations.A chronic infection of immunocompetent cells is most likely at the origin of a benign mono-oligoclonal
B lymphocyte proliferation, typically observed in mixed cryoglobulinemia(10% showing late B-NHL).
Aim. Identify early markers of autoimmune lymphoproliferative disease onset in a group of antiviral treatment-naïve patients
infected by HCV that could identify the transition between a state of
silent autoimmune and lymphoproliferative conditions and frank
disease.
Material, Methods and Results.Fourty patients were
recruited. Antinuclear antibodies(ANA) were detected by indirect
immunofluorescence. Autoantibody detection of IgG directed
against M2,gp210,SP100,LKM1,LC1,SLA,Factin antigens were performed by Immunodot analysis. Free light chain(FLC) detection
were carried out by turbidimetric assay. Cryoglobulin and cryofibrinogen analysis was carried out following the guidelines of the
SIBIOC. Our results show an 84% prevalence of cryoglobulinemia
in samples collected from HCVinfected patients. Of these, 27%
showed ANA positivity a negligible percentage of autoantibody
liver disease and absence of positivity of cryofibrinogen. The most
significant result concerns the finding of high doses of FLC in
73% of patients, of which 21% showing an abnormally elevated
k/l ratio. Statistical analysis suggests that patients presenting
cryoglobulinemia and FLCratio above 1.6 are also ANApositive.
Conclusions. ANA positivity is indicative of the presence of a
persistent antigenic stimulus by the virus and the activation of any
autoimmune clones.The presence of cryoglobulinemia suggests a
continuous lymphocyte stimulation. Interestingly, our results suggest a possible role for the presence of high levels of FLCs and their
use to identify the transition between a silent state of probable
autoimmune lymphoproliferative disease or a frank illness, using
k/l ratio as a cut-off value. The presence of a subpopulation of HCVpositive patients may open new scenarios to targeted therapeutic
treatment strategies in subclinical phases. Our study is a contribution to presenting a panel of potential predictive markers of disease
progression.
e57
Padua, Italy
Background and aim: Ascites represents the hallmark of
decompensation in patients with cirrhosis. Apart from the assessment of HVPG, that is not yet part of clinical practice, predictors
of ascites onset are lacking, restricting the adoption of preventive
strategies. The presence of a modest layer of fluid (MLF) around
the liver and/or the spleen at ultrasound examination is a frequent
finding in patients with compensated cirrhosis. The meaning of MLF
has never been investigated in these patients. Thus, the aim of this
study was to evaluate if the MLF precedes the appearance of ascites.
Methods: 248 consecutive patients with cirrhosis without
ascites, were enrolled and followed up until death, liver transplantation and/or for a maximum of 10 years.
Results: Ninety-three patients (37.5%) had MLF at inclusion.
Patients with MLF were older (58.3vs50.8 years; p < 0.001), had
a higher MELD score (11.8vs10.7; p = 0.028), a higher ChildPugh score (7.4vs6.4; < 0.001) and a lower mean arterial pressure
(92.1vs97.4; p < 0.001) than patients without MLF. Ten-year probability to develop overt ascites was significantly higher in patients
with MLF than without MLF (38.3vs22.7%;p < 0.001). MLF was found
to be an independent predictor of the development of over ascites
(hazard ratio[HR] = 2.2; p = 0.023) as well as albumin (HR = 0.86;
p < 0.001) and treatment with ␤-blockers (HR = 2.29; p = 0.024).
Conclusions: The presence of MLF and the use of ␤-blockers
without a verfied effect on HVPG were found to be the strongest
predictors of ascites onset in patients with cirrhosis. MLF is an easily
accessible predictor of the appearance of overt ascites in patients
with cirrhosis. Thus, it can be used to plan preventive interventions
such as a dietary salt restriction and/or the early prescription of an
antialdosteronic drug.
e58
F-32
F-33
CHARACTERISTICS OF PATIENTS WITH CHRONIC
HEPATITIS C WHO HAVE FAILED PAST
TREATMENT OR WHO HAVE NEVER BEEN
TREATED: HOW MUCH ROOM LEFT FOR
INTERFERON?
ELF TEST IS A RELIABLE TOOL FOR
NON-INVASIVE DIAGNOSIS OF LIVER FIBROSIS IN
PATIENTS WITH NONALCOHOLIC FATTY LIVER
DISEASE
A. Ciaccio 1 , S. Okolicsanyi 1 , M. Rota 2 ,
P.A. Cortesi 3 , S. De Salvia 1 , M. Gemma 1 ,
M. Vinci 4 , L.G. Mantovani 3 , L.S. Belli 4 ,
M. Strazzabosco 1,5
1
Department of Surgery and Translational Medicine,
Milano-Bicocca, Milan, Italy
of Milano-Bicocca, Milan, Italy
4 Department of Hepatology and Gastroenterology,
Liver Unit, Niguarda Hospital, Milan, Italy
Background/aims: According to current guidelines on management of Chronic Hepatitis C (CHC), both interferon (IFN)-containing
and IFN-free treatment combinations may be offered to patients,
depending on viral genotype, previous treatment and contraindications to IFN. The past two decades of treatment have selected a
large population of patients ineligible to IFN. Our aim is to characterize the current CHC population in order to best drive resource
allocation.
Patients and methods: we retrospectively retrieved the characteristics and outcomes of a cohort of 801 consecutive CHC
outpatients followed-up at two tertiary Centers in Northern Italy.
Baseline characteristics of never treated and treated CHC patients
were compared by means of Chi-Square test for categorical and
t-test for continuous variables.
Results: fifty-three percent (426 out of 801) of patients had previously received treatment. Among them, 103 (24%) had achieved
SVR; 323 (76%) had failed, of whom 74 (23%) did not complete
and 249 (77%) did not respond to treatment. Adverse events were
the main cause of treatment withdrawal (90%). Forty-seven percent (n = 375) of all participants had never received any treatment,
mostly because of contraindications (27% overall, hematologic 11%,
psychiatric 24%, cardiovascular 15%, autoimmune 7%, others 43%),
advanced age (27%), personal choice (15%) or no fibrosis (10%).
Never treated patients differed significantly (p < 0.01) from treated
ones by age (61 vs 56 years) and gender (male 49 vs 65%). Sixhundred and ninety-eight patients (323 failed plus 375 never
treated) are awaiting new antiviral therapy, of which 442 (63%) are
candidate to all-oral combinations because of contraindications or
past intolerance to IFN, while the remaining 256 (37%) may still
receive an IFN-containing regimen.
Conclusions: 37% of the CHC patients being followed in our clinics may receive IFN-containing protocols with newer DAAs, while
about the two thirds of these patients are obligated candidates for
the more costly, IFN-free regimens.
L. Miele 1 , M.A. Isgrò 2 , T. De Michele 2 ,
G. Marrone 1 , C. Cefalo 1 , S. Racco 1 , L. Viti 1 ,
A. Giannace 2 , C. Morlacchi 2 , G.L. Rapaccini 1 ,
A. Gasbarrini 1 , C. Zuppi 2 , A. Grieco 1
1 Department of Internal Medicine, Catholic
2 Department of Laboratory Medicine, Catholic
Introduction: Recently, the serum Enhanced Liver Fibrosis (ELF)
Test has been developed for staging liver fibrosis in patients with
chronic liver diseases.
Aim: The aim of our study was to evaluate the ELF Test performance in predicting fibrosis stage in an independent adult cohort
of NAFLD patients.
Materials and methods: 82 patients (mean age 46 years)
with suspected NAFLD were enrolled undergoing percutaneous liver biopsy and serum sampling. Fibrosis was assessed
and scored by using the modified Brunt classification (F0 = no
fibrosis; F1 = perisinusoidal/periportal; F2 = perisinusoidal and portal/periportal; F3 = bridging fibrosis; F4 = cirrhosis). The ELF test
was determined in all patients by means of an algorithm combining
hyaluronic acid, amino-terminal propeptide of type III collagen and
tissue inhibitor of metalloproteinase 1. Diagnostic accuracy was
assessed determining the area under receiver operating characteristic curves (AUCs).
Results: The distribution of fibrosis stages in our cohort was
as follows: F0 = 7.3% (n = 6), F1 = 39.0% (n = 32), F2 = 35.4% (n = 29),
F3 = 6.1% (n = 5), F4 = 12.2% (n = 10). The ELF Test had an AUC of 0.988
(95% Confidence Interval C.I. 0.967-1.008; P < 0.001) for distinguishing cirrhosis, 0.948 (C.I. 0.883-1.014; P < 0.001) for severe fibrosis,
0.682 (C.I. 0.568-0.797; P = 0.005) for significant fibrosis and 0.658
(C.I. 0.401-0.915; P = 0.200) for any fibrosis. ELF scores were significantly higher in patients with severe fibrosis/cirrhosis in respect
to ones with no/mild/moderate fibrosis (median 11.26 vs. 8.53;
P < 0.001). Severe fibrosis and cirrhosis were correctly identified in
91% of patients.
Conclusions: In our cohort of NAFLD patients, the ELF test was
able to discriminate severe fibrosis and cirrhosis with an excellent
diagnostic accuracy. It may result useful for the selection of cases
with more advanced fibrosis stage and for therapeutic follow-up,
thus avoiding liver biopsy.
F-34
F-35
IMPACT OF ANTICOAGULATION ON THE
EVOLUTION OF PORTAL VEIN THROMBOSIS IN
CIRRHOSIS: A RETROSPECTIVE ANALYSIS
SERUM SCCA-IGM CAN PREDICT THE
DEVELOPMENT OF HCC IN PATIENTS WITH
HCV-CIRRHOSIS
I. Pettinari 1 , S. Landi 1 , S. Berardi 2 , C. Serra 2 ,
P. Pianta 2 , P. Caraceni 3 , G. Verucchi 4 ,
M. Cescon 5 , M.R. Tamè 6 , L. Bolondi 1 ,
F. Piscaglia 1
G. Pieri 1 , S. Marenco 1 , F. Lantieri 2 ,
I. Baldissarro 1 , L. Bruzzone 1 , V. Fazio 1 ,
S. Labanca 1 , G. Sammito 1 , V. Savarino 1 ,
A. Picciotto 1
1
1
Unit of Internal Medicine, Department of Digestive
Disease, General and University Hospital
S.Orsola-Malpighi, Bologna, Italy
2 Unit of Internal Medicine and Organs Failure,
General and University Hospital S.Orsola-Malpighi,
Bologna, Italy
3 Unit of Semeiotica, Department of Digestive
4 Unit of Infectious Disease, General and University
Hospital S.Orsola-Malpighi, Bologna, Italy
5 Unit of Hepatobiliary Surgery, Department of
Organs Failure and Transplantations, General and
University Hospital S.Orsola-Malpighi, Bologna, Italy
6 Unit of Gastroenterology, Department of Digestive
Background: Non-neoplastic portal vein thrombosis (PVT) is
a common complication in cirrhosis, however no consensus exist
about any management and therapeutical algorithm for this condition.
Aim of our study was to investigate the evolution of PVT in
cirrhosis and the influence of anticoagulant treatment.
Materials and methods: A retrospective analysis of 68 cirrhotic
patients with non-malignant PVT observed at the S.OrsolaMalpighi General and University Hospital from January 2008 to
December 2013 was performed. Patients with HCC were not
excluded provided that PVT was ascertained as non neoplastic.
Thrombosis diagnosis and assessment of recanalization were performed using Doppler ultrasound, spiral CT or MRI. Anticoagulants
were administered to only 25 of 68 patients at thrombosis detection. Median follow- up was 24 months in treated and 17 months
in untreated patients.
Results: In the 43 patients not receiving anticoagulant therapy, PVT remained stable in 21 (48.8%), progressed in 15 (34.8%), 8
of whom subsequently started anticoagulants and spontaneously
improved in 7 patients (16.4%). Recanalization was achieved in 16 of
25 patients (64%) receving anticoagulation (11 complete and 5 partial PVT), while no recanalization was observed in 9 anticoagulated
patients (36%). Seven of 11 patients who had achieved complete
recanalization suffered PVT recurrence after stopping anticoagulation. Bleeding, but not lethal complications occurred in 7 treated
patients (28%), of which 3 were suspected to be anticoagulationrelated.
Conclusion: Our study confirms that anticoagulation therapy influences the course of PVT, favoring recanalization and
preventing thrombus extension. However discontinuation of anticoagulation therapy is associated with high risk of recurrent
thrombosis. Anticoagulant treatment appeared to be relatively safe.
e59
U.O.s. Diagnosi e Terapia delle Epatopatie, IRCCS,
San Martino IST, Genova, Italy
2 DISSAL, Biostatistics Dep., University of Genoa, Italy
Introduction and aim. The serum level of circulating squamous cellular carcinoma antigen (SCCA)-IgM immunocomplex has
been proposed as marker for diagnosis of hepatocellular carcinoma
(HCC) and its progressive increase is associated with development
of tumor. We studied the role of a single SCCA-IgM measurement
in predicting the development of HCC.
Material and Methods. We retrospectively selected 93 patients
with cirrhosis (26 with HCC and 67 without HCC at enrollment). For
each patient, we chose serum samples collected at least 6 months
before the diagnosis of HCC or at least 12 months before the enrolment. We measured SCCA-IgM using an ELISA assay kit. We applied
the ROC-curve for the best cut-off; independent Student’s t-test
and Fisher’s test for parametric data; Mann-Whitney U-test for
non-parametric data; Kaplan-Meier to test the survival.
Results. There were no differences for age, gender and HCVinfection between the groups with and without HCC. The best
SCCA-IgM predictive value of HCC development was 192 AU/mL
(sensibility 65.4%, specificity 62.7%, AUROC 0.607). SCCA-IgM levels were higher in HCV versus non-HCV patients (p = 0.027), both
in HCC and HCC-free group. Considering the HCC occurrence and
the HCV etiology together (ANOVA), the second one was better
related to SCCA-IgM (p = 0.04). SCCA-IgM levels > 192 AU/mL were
more frequent in patients who developed HCC than in pantients
who did not develop HCC (p = 0.032). The predictivity of SCCAIgM levels increased considering male HCV-positive patients alone
(sensitivity 85.7%, specificity 57.1%, AUC 0.707) with the same cutoff. The survival analysis revealed that the mean HCC-free survival
in patients with SCCA-IgM <192 AU/mL was longer than in patients
with SCCA-IgM 3 192 AU/mL (105.1 vs 84.7 months, p = 0.024).
Conclusions. SCCA-IgM levels are higher in HCV-positive
patients and can help identifying those with higher risk of earlier
development of HCC, using a cut-off of 192 AU/mL.
F-36
ANTIHBS IS A USEFUL TOOL FOR SEARCHING THE
RISK OF HEPATITIS B REATIVATION: A
META-ANALYSIS OF OBSERVATIONAL STUDIES
IN PATIENTS WITH LYMPHOMA
M. Fiore 1 , I. Baldi 2 , A. Floreani 3
1 Infectious Disease Unit, University Hospital of
Trieste, Trieste, Italy
2 Department of Cardiac, Thoracic and Vascular
Sciences, University of Padua, Padua, Italy
3 Department of Surgical, Oncology and
Background: Reactivation of HBV is a well-recognized complication following immunosuppression in patients with past
infection. The International guidelines (AASLD, EASL) recommend
e60
detection of antiHBc regardless of antiHBs status in subjects undergoing chemotherapy, and prophylaxis in those with viral load
detectable.
Aim: To perform a meta-analysis of papers dealing on this
issue searching for two primary outcome measures: i) the proportion of reactivation of HBV in HBsAg-ve patients according to the
antiHBc status; ii) the proportion of reactivation of HBV in HBsAgve/antiHBc + ve patients according to the antiHBs status.
Methods: We searched the PuMed collection from any date to
February 28, 2014 for the following terms: “HBV, hepatitis B, reactive. Of the 1,226 screened papers only 11 papers fulfilling the
inclusion criteria were included (full papers). The Der Simonian
and Laird effects models together with the Duval and Tweedie non
parametric “trim and fill” method of accounting for publication bias
were used.
Results: Patients positive for antiHBc alone showed a 4%
increased risk of reactivation of HBV which was significantly higher
than antiHBc-ve subjects (95%CI: 1-6%, p = .002). The asymmetry of
funnel plot analysis suggested the presence of a publication bias,
thus the risk differences were reduced to 2.3% (p = .09) according to
the “trim and fill” analysis. Considering the second endpoint measure, a significant reduction of the risk of reactivation was obtained
pooling antiHBs + ve and antiHBs-neg subjects (-8%, 95%CI:-12 to
-4%, p < .001). Finally, based on these data, an algorithm for HBV
prophylaxis was constructed, recommending antiHBs searching as
a useful tool for management of HBV reactivation.
Conclusion: The antiHBs assay may reduce the risk for HBV
reactivation and HBV-associated morbidity/mortality in patients
with past HBV infection undergoing chemotherapy for lymphoma.
F-37
“EARLY” SUBCLINICAL LEFT VENTRICULAR
DISFUNCTION IN PATIENTS WITH HEPATITIS C
VIRUS INFECTION
A. Rocco 2 , M. Sanduzzi Zamparelli 2 ,
D. Angrisani 2 , D. Compare 2 , C. Santoro 2 ,
M. Galderisi 1 , G. Nardone 1
1 Hypertension Research Center and Department of
Medical Translational Sciences, Federico II University
of Naples, Naples, Italy
Gastroenterology Unit, Federico II University of
Introduction: Hepatitis C virus (HCV)infection has been associated with several extrahepatic manifestations in a variety of tissues
and organs, including the myocardium. Reduced myocardial perfusion or increased pro-B-type natriuretic peptide (NT-proBNP)
have been described in HCV-infected individuals but, until now,
no study clearly demonstrated the occurrence of myocardial dysfunction in the early stages of HCV-related liver diseases.
Aim: to evaluate the impact of HCV infection on left ventricular
(LV) geometry and function by a standard 2D and 3D echocardiography.
Material and Methods: Thirty consecutive patients with histologically proved HCV-related chronic hepatitis and twenty normal
controls (NCs), comparable for age and gender prevalence were
enrolled in the study. Exclusion criteria were: other causes of
liver diseases, coronary artery and/or valvular heart disease, heart
failure, cardiomyopathies, atrial fibrillation or alcohol abuse. The
echocardiography protocol consisted of: standard echo Doppler
plus LV volumetric and systolic function analysis through 3D
echocardiography. Left ventricular systolic function thorough 3D
echo was evaluated by STE derived global longitudinal strain (GLS),
global circumferential strain (GCS), global area strain (GAS) and
global radial strain (GRS).
Results: Overall we enrolled 16 males and 14 females with HCV
(genotype 1) related chronic liver disease. Mean age was 60.9 ± 6.5
yrs. NCs were comparable for age, gender distribution, body mass
index, heart rate and blood pressure. In HCV infected subjects
standard 2D echocardiography point out a transmitral E/A ratio
marginally lower (0.84 ± 0.2) compared to NC (1.01 ± 0.3) (p = 0.04).
Moreover, more detail 3D STE showed a subclinical alteration of
LV function documented by lower GCS (HCV = -15.3 ± 2.2%, NC = 17.6 ± 3.9%, p = 0.02), GAS (-25.2 ± 2.6%, NC -29.9 ± 5.2%, p = 0.03),
and GRS (37.6 ± 4.6% vs 44.9 ± 12%, p = 0.04) whereas HCV-releated
changes of GLS were not significant.
Conclusion: HCV chronic infection is associated a subclinical
myocarditis-like damage documented by the alteration of the circumferential fibers of the ventricular mid wall.
F-38
OXIDATIVE STRESS, MITOCHONDRIA DAMAGE
AND MATRIX METALLOPROTEASE ACTIVATION
IN THE PATHOGENESIS OF NAFLD
C. Berardo 1,∗ , G. Palladini 1 , L.G. Di Pasqua 1 ,
V. Rizzo 2 , S. Perlini 1 , P. Richelmi 1 , M. Vairetti 1 ,
A. Ferrigno 1
1 Department of Internal Medicine and Therapeutics,
Fondazione IRCCS Policlinico S. Matteo, University of
Pavia, Pavia, Italy
2 Department of Molecular Medicine, Fondazione
IRCCS Policlinico S. Matteo, University of Pavia,
Pavia, Italy
Background and Aims: Animal models of hepatic steatosis
have allowed the understanding of mechanisms involved in the
pathogenesis of non-alcholic fatty liver disease (NAFLD). By comparing these models a working hypothesis for NAFLD pathogenesis
emerges as well as the mechanism implicated in the progress to
nonalcoholic steatohepatitis (NASH). This study investigated the
oxidative stress, mitochondria damage and matrix metalloprotease
activation in fatty livers using two rat models of NAFLD such as the
methionine and choline deficient (MCD) diet model and obese fa/fa
rats.
Methods. Male Wistar rats underwent to NAFLD induced by 4week MCD diet; blood samples and hepatic biopsies were collected
up to 4 weeks. 12-week male old obese (fa/fa) and lean (fa/-) male
Zucker rats were also used. Serum hepatic enzymes (AST, ALT, LDH
and g-GT) and total and direct bilirubin were quantified. Tissue
glutathione (GSH), lipid peroxides, ATP/ADP ratio and matrix metalloproteinase activation (MMP-2, MMP-9) were also evaluated.
Results: No significant changes in serum AST, ALT, LDH and gGT were found comparing the two animal models. A marked serum
increase in total and direct bilirubin at IV weeks in MCD rats was
detected. Mostly decrease in tissue GSH levels and increase in the
lipid peroxides were found in the MCD group. A better mitochondrial function as documented by the ATP/ADP ratio and no MMP-9
and poor MMP-2 activation was measured in obese fa/fa rats when
compared with the MCD animals. Instead, in MCD rats a significant
increase of MMP2 activation occurred.
Conclusions: MCD rats exhibit a marked oxidative stress and
mitochondria damage concomitant with an MMP-2 activation. The
reported spontaneous development to severe NASH observed only
in MCD rats and not in obese fa/fa animals might be associated to
these events already detectable in the early period of treatment.
Funding: Supported by Fondazione Cariplo, grant n◦ 2011-0439.
F-39
Onorato 1 ,
N.
L.
C.
E. Sagnelli 1 , I.F. Angelillo 3
F-40
A STRATEGY TO FAVOR THE ACCESS OF
IRREGULAR AND REFUGEE MIGRANTS TO A
SCREENING PROGRAM FOR HBV, HCV AND HIV
INFECTION
N. Coppola 1 , L. Alessio 1,2 , L. Gualdieri 3 ,
M. Pisaturo 4,5 , C. Sagnelli 2,6 , N. Caprio 1,7 ,
R. Maffei 1,3 , M. Starace 1 , I.F. Angelillo 8 ,
G. Pasquale 1 , E. Sagnelli 5
HBSAG NEGATIVE ANTI-HBC POSITIVE
SEROLOGY INCREASES THE RISK OF HCC IN
CHRONIC HEPATITIS PATIENTS: A
META-ANALYSIS
Coppola 1 ,
e61
Sagnelli 2 ,
Italy
3 Department of Experimental Medicine, Second
Background and Aims: We performed a meta-analysis to ascertain if patients with chronic hepatitis (CH) of different etiology
and anti-HBc positive are at a higher risk to develop hepatocellular
carcinoma (HCC) than those anti-HBc negative.
Methods: studies meting these criteria were included:
investigating on the relationship between HBsAg-negative/antiHBc-positive serology and occurrence of HCC, being a case-control
or cohort study, providing relative risk or odds ratios and 95% confidence intervals, being available as full text written in English, being
published and indexed up to September 2014.
Results: Twenty-six studies met inclusion criteria, allowing a
meta-analysis on 14,558 patients. Table 1 shows the results of the
meta-analysis
Conclusions: This meta-analysis shows that in HBsAg negative
CH patients anti-HBc positivity is strongly associated with HCC, an
association observed in all subgroups established by the stage of
the disease, aetiology and ethnicity
2 Medical Center, Centro Sociale ex Canapificio,
Caserta, Italy
3 Medical Center, Centro per la Tutela della Salute
degli Immigrati, Naples, Italy
4 Medical Center, Centro di Accoglienza “La tenda di
Abramo”, Caserta, Italy
5 Infectious Diseases Unit, AORN Sant’Anna e San
Sebastiano, Caserta, Italy
Italy
7 Medical center, Centro Suore Missionarie della
Carità, Naples, Italy
8 Department of Experimental Medicine, Second
Background: Half immigrants are irregular or refugee frequently excluded from social life by linguistic, cultural and
socioeconomic barriers also limiting their access to the healthcare
services.
Aim: To analyze the use of strategies favoring the access of
irregular or refugee migrants to a screening program for HBV, HCV
and HIV infection.
Methods: A screening for HBV, HCV or HIV infection was proposed to 926 migrants by a team (doctors, nurses and cultural
mediators) skilled in managing problems of vulnerable groups. The
screening, free of charge and with no bureaucracy, was accepted by
882 (95.2%) migrants, 625 irregular and 257 refugee, and performed
Table 1
Meta-analysis data on the development of HCC in CH anti-HBc positive or anti-HBc negative.
HCC in
No of studies
N◦ of patients
Anti-HBc + /anti-HBc-
N◦ and (%) HCC
Anti-HBc+/anti-HBc-
RR (efficacy)
95% CI (efficacy)
p
Heterogeneity test
(Q;p;I2 ,%)
All subjects
chronic liver diseases*
Chronic hepatitis
Liver cirrhosis
Asian subjects
Non-Asian subjects
Anti-HCV positive subjects
Anti-HBs positiveˆ
Anti-HBs negativeˆˆ
Anti-HBs positiveˆˆ
26
23
3
7
16
10
20
9
9
9
5581/8977
4614/5034
370/408
973/959
3639/4230
1942/4747
4747/8082
935/4198
719/4198
935/719
1705 (30.5)/1257 (14.0)
1513 (32.8)/1119 (22.2)
76 (20.5)/62 (15.2)
320 (32.9)/196 (20.4)
1231 (33.8)/859 (20.3)
474 (24.4)/398 (8.4)
1348 (28.4)/999 (12.4)
223 (23.8)/384 (9.1)
331 (46.0)/384 (9.1)
223 (23.8)/331 (46.0)
1.58
1.45
1.44
1.30
1.62
1.51
1.52
1.17
1.75
0.63
1.49-1.69
1.36-1.54
1.18-1.77
1.12-1.52
1.50-1.74
1.34-1.69
1.4-1.63
1.01-1.35
1.56-1.96
0.55-0.72
0.000
0.000
0.000
0.001
0.000
0.000
0.000
0.03
0.000
0.000
99.91;0.000;75.0
101.62;0.000;75.4
1.16;0.56;0.0
10.85;0.093;44.7
34.51;0.003;56.5
65.14;0.000;86.2
64.62;0.000;72.1
43.78;0.000;81.7
120.61;0.000;93.4
18.61;0.017;57.0
* Excluding the healthy controls. ˆ Anti-HBs/anti-HBc positive subjects vs. anti-HBs/anti-HBc negative subjects. ˆˆ Anti-HBs negative/anti-HBc positive subjects vs. antiHBs/anti-HBc negative subjects. ˆˆˆ Anti-HBs/anti-HBc positive subjects vs. anti-HBs negative/anti-HBc positive patients.
e62
at 4 territorial medical centers in Campania region, southern
Italy.
Results: 78 (8.7%) migrants were HBsAg positive, 35 (3.7%)
anti-HCV positive, 11 (1.3%) anti-HIV positive and 7 (0.8%) had a
multiple infection. Of the 801 HBsAg negative patients, 373 (40.2%)
were anti-HBc positive. All migrants with a detectable marker of
HBV, HCV or HIV infection were unaware of their serological status. The HBsAg positivity rate was high (13.9%) in migrants from
sub-Saharan Africa and intermediate in those from eastern Europe
(6.1% of 194 cases), northern Africa (2.5% of 80 cases) and IndianPakistani area (3.2% of 126 cases). Anti-HCV was more frequent
in migrants from eastern Europe (6.2%) and Indian-Pakistani areas
(7%) than in those from sub-Saharan (3.8%) and northern Africa
(2.5%). Anti-HIV was detected in 17 (3.8%) migrants from subSaharan Africa, in 5 (2.4%) from eastern-Europe, in 1 (0.8%) from
Indian-Pakistani area and in none from northern Africa.
Discussion. This investigation identified 131 migrants with
HBV, HCV or HIV infection unaware of their serological status, suggesting the need of a strong action of Healthcare Authorities of Italy,
may be using our model, to favor the access of migrant populations
to healthcare services and to support screening and educational
programs
F-41
LAMIVUDINE PROPHYLAXIS PREVENTS
HEPATITIS B REACTIVATION IN
HBSAG-NEGATIVE/ANTI-HBC-POSITIVE
PATIENTS UNDERGOING RITUXIMAB-BASED
CHEMOTHERAPY FOR NON–HODGKIN’S B CELL
LYMPHOMA
M. Viganò 1 , G. Grossi 2 , E. Borsotti 2 ,
M. Cappelletti 1 , M. Goldaniga 3 , L. Farina 4 ,
M. Rumi 1 , P. Corradini 4 , L. Baldini 3 ,
M. Colombo 2 , P. Lampertico 2
3 Department of Oncohematology, Fondazione IRCCS
Cà Granda Ospedale Maggiore Policlinico, University
of Milan, Milan, Italy
4 Division of Hematology, IRCCS Istituto Nazionale
Tumori, University of Milan, Milan, Italy
Introduction and Aim: Hepatitis B surface antigen (HBsAg)negative/anti-hepatitis B core antigen (anti-HBc)-positive patients
undergoing Rituximab (RTX)-based chemotherapy (R-CT) for
non–Hodgkin’s B cell lymphoma (NHL) face up to 27% risk of hepatitis B virus (HBV) reactivation. Aim of the study was to assess
the efficacy and safety of lamivudine (LMV) prophylaxis in these
patients.
Materials and Methods, Results: Sixty-seven consecutive
HBsAg-negative/anti-HBc-positive patients (median age 70 yrs,
61% males, 13% HCV seropositive, 100% serum HBV DNA negative
by sensitive PCR assay, 75% anti-HBs positive, 100% with ALT < ULN)
undergoing different R-CT protocols (48% R-CHOP) in two hematological centres were retrospectively enrolled. LMV prophylaxis was
started before the first R-CT dose and planned to last for 18 months
after the end of R-CT. R-CT was administered for 6 cycles (range: 115) during 5 (range: 1-38) months, while LMV was administered for
20 (range: 3-54) months. Serum ALT, HBsAg and HBV DNA levels by
PCR assay were assessed every 3-4 months. All patients remained
HBV DNA and HBsAg negative during a median of 20 months (range:
2-60), including both the LMV prophylaxis and the subsequent off
treatment follow-up. Anti-HBs titers declined in 18% of patients
but none lost serum reactivity for anti-HBs. Overall, 2 patients had
an increase of ALT (> ULN) but this event was unrelated to HBV
reactivation and 7 patients died of liver unrelated causes. No safety
issues related to LMV were recorded.
Conclusions: LMV monotherapy is an inexpensive, safe and
effective prophylaxis regimen to prevent HBV reactivation in such a
high-risk population as HBsAg-negative/anti-HBc-positive patients
receiving RTX-based chemotherapy for non–Hodgkin’s B cell lymphoma.
F-42
HBV GENETIC COMPARTIMENTALIZATION,
VARIABILITY AND MOLECULAR CORRELATES OF
HISTOLOGIC AND IMMUNOHISTOCHEMICAL
ASPECTS IN LIVER TISSUE: IMPLICATIONS FOR
THE CLINICAL MANAGEMENT OF PATIENTS
WITH CHRONIC HEPATITIS B
C. Minosse 1 , A. Baiocchini 2 , M. Selleri 1 ,
E. Giombini 1 , S. Coen 1 , P. Zaccaro 1 , G. Rozera 1 ,
D. Vincenti 1 , F. Del Nonno 2 , U. Visco Comandini 3 ,
R. Lionetti 3 , M. Montalbano 3 , G. D’Offizi 3 ,
M. Vivarelli 4 , M.R. Capobianchi 1 , S. Menzo 5
1 Laboratory of Virology, National Institute for
Infectious Diseases “L. Spallanzani”, Rome, Italy
2 Laboratoy of Pathology, National Institute for
3 1st Clinical Division, National Institute for
4 Hepatobiliary and Transplantation Surgery Unit,
Università Politecnica delle Marche, Ancona, Italy
5 Laboratory of Virology, Università Politecnica delle
Marche, Ancona, Italy
Introduction In chronic hepatitis B (CHB), viral intrahepatic
molecular (ccc- and total DNA) and immunohisto chemical markers should allow the best representation of viral biology and organ
pathology. Presently, the role of these intrahepatic markers in the
pathogenesis of CHB and their clinical significance are still unclear.
Furthermore, the the genetic determinants of intrahepatic HBV
(compared to plasma), have essentially never been considered.
Aim This study focused on the integrated analysis of intrahepatic molecular and histochemical markers to elucidate some
pathogenetic mechanisms underlying chronic HBV infection and
disease.
Materials and Methods, Results The study enrolled 35 consecutive untreated CHB patients. In addition to classical virological and
clinical parameters, intrahepatic cccDNA, total DNA quantification
(by real-time PCR) and intrahepatic viral antigen distribution (by
immunohistochemistry) were evaluated in liver tissue. Conventional and next generation sequencing of the pre-core and pre-s
regions of viral genome was performed on all viral compartments.
Precore mutations were shown to be responsible of cytoplasmatic accumulation of HBsAg and reduced viral replicative capacity
(totalDNA/cccDNA). A missense precore deletion was associated
to G3 steatosis in one patient. Mainstream intrahepatic virus was
absent from plasma in 2 patients. Other patients displayed moderate compartimentalization. Interestingly, variants not appearing in
the peripheral blood carried missense indels in the precore region,
driving the synthesis of abnormal peptides, possibly leading to
virion assembly defects and to the intracytoplasmatic accumulation of HBsAg.
Conclusions Selective pressure by the immune system selects
variants in the precore region that are not proportionally detectable
in the periphery, possibly due to the abnormal release of defective
virions. This may also have clinical implications for hepatic pathology (steatosis) and may lead to the underestimation of viral load. A
better comprehension of the relationship between intrahepatic and
peripheral (virological and clinical) markers may confer to these a
higher diagnostic power.
F-43
SARCOPENIA AT FIRST DIAGNOSIS PREDICTS A
REDUCED SURVIVAL IN PATIENTS AFFECTED BY
HEPATOCARCINOMA
G. Antonelli 1 , P. Begini 1 , E. Gigante 1 ,
F. Carbonetti 2 , E. Iannicelli 2 , S. Gallina 1 ,
A. Pellicelli 3 , L. Miglioresi 3 , G. Delle Fave 1 ,
M. Marignani 1
1 Digestive and Liver Diseases Dpt., Sant Andrea
Hospital, School of Medicine and Psychology,
Sapienza University, Rome, Italy
2 Radiology Dpt., Saint Andrew Hospital, School of
Medicine and Psychology, Sapienza University,
Rome, Italy
3 Liver Unit., San Camillo Forlanini Hospital, Rome,
Italy
Introduction Sarcopenia is a frequent complication and an
independent risk factor for mortality and clinical outcomes in
patients (pts) with liver cirrhosis and in a variety of other clinical
conditions.
Aim of the study was to determine the prevalence of sarcopenia
in a cohort of cirrhotic pts at the first diagnosis of hepatocellular
carcinoma (HCC) treated in a tertiary center and its influence on
survival.
Methods All consecutive HCC pts treated at our outpatient clinic
(years 2004-2014) undergoing abdominal computed tomography
(CT) were retrospectively studied with a software analyzing the
cross-sectional areas of muscles at L3 level. Data was normalized
for height obtaining the Skeletal Muscle Index (SMI) to measure
sarcopenia. Presence of sarcopenia was defined when SMI was
≤41 cm2/m2 for women and ≤53cm2/m2 for men with a body mass
index (BMI) ≥25, and ≤43 cm2/m2 for men with BMI < 25.
Results 92 HCC pts were evaluated [27F (29.4%), 65 M (70,6%].
Age at diagnosis was 71,9 years (30,7-86,4), while BMI was 24,7
(17,5-36,7). Viral infection was the cause of liver disease in most
cases. Distribution by CHILD score was as follows: A = 55,4%;
B = (42,4%); C = (2,2%). Median MELD score was 10 (6-18). Metastatic
disease was present in 12% of cases. The distribution by BCLC was as
follows: A = (41,3%);B = (25%), C = 28,3%, D = 5,4%. Overall, sarcopenia was present in 40,2%. Baseline features were similar between
sarcopenic vs non sarcopenic pts. Sarcopenic pts were prevalently
females (p = 0,0044) and had a reduced mean Overall Survival of
123 (95% C.I. 98 to 150) vs 66 (95% C.I. 47 to 84) weeks (p = 0,001).
Multivariate analysis has shown that sarcopenia was indipendent
of age (p = 0,0027).
Conclusions The prevalence of Sarcopenia in HCC pts is high,
with a greater frequency in female pts. Presence of Sarcopenia at
diagnosis of HCC is an important predictor of a reduced survival.
e63
F-44
PANTOPRAZOLE AND RABEPRAZOLE DO NOT
IMPACT THE ACTIVITY OF CYTOCHROME P450
ASSESSED BY [13 C]-AMINOPYRINE BREATH
TEST IN PATIENTS WITH LIVER CIRRHOSIS
A. Rocco 2 , D. Angrisani 2 , C. Rubicondo 2 ,
L. Staiano 1 , D. Amoruso 1 , D. Compare 2 ,
C. Coppola 1 , G. Nardone 2
1
Hepatology and Interventional Ultrasound Unit,
Gragnano Hospital, Gragnano, Italy
Gastroenterology Unit, Federico II University of
Naples, Italy
Introduction: Proton pump inhibitors (PPIs) are one of the most
widely used drugs worldwide. Almost all PPIs undergo extensive
hepatic metabolism via cytochrome (CYP)-P450 system, specifically CYP-2C19 and CYP-3A4. Pantoprazole is metabolized through
the CYP-P450 system, while the primary pathway of rabeprazole
metabolism is non enzymatic. In advanced liver diseases the activity of drug metabolizing enzymes is impaired, thus increasing
the risk of drug accumulation, drug-drug interaction and adverse
events.13 C-aminopyrine breath test (13 C-ABT) is a noninvasive,
liver function test that explores CYP enzyme activity.
Aim: to evaluate the effects of different PPIs on the activity of
CYPs by 13 C-ABT in patients with Hepatitis C virus (HCV)-related
liver cirrhosis.
Material and Methods: Thirty consecutive genotype 1 HCVrelated liver cirrhosis patients, Child-Pugh A, were randomly
assigned to pantoprazole (40 mg/day) or rabeprazole (20 mg/day).
13 C-ABT was performed before and 15 days after starting PPI
by collecting breath samples baseline and at 30-minute intervals for 2 hours after oral administration of 13 C-aminopyrine
(2 mg/Kg body weight). 13 C-enrichment of CO2 was determined
by purification-isotope ratio mass spectrometer. Results were
expressed as maximum percentage of 13 CO2-recovery per hour
(max 13 C% dose/h) at any time (“excretion peak”) and percentage of 13 CO2-cumulative dose recovered in 2 h (%13 C cum dose at
120 min).
Results: Overall, we enrolled 13 males and 17 females. Mean
age was 60.9 ± 7.5 yrs. Age, gender distribution, BMI and laboratory findings did not significantly differ among pantoprazole and
rabeprazole group. Baseline, 13 C-ABT results were altered in 13/30
(43%) patients (6 in pantoprazole and 7 in rabeprazole group). Fifteen days after PPIs, 13 C-ABT did not significantly changed in terms
of both %dose/h and %dose/cumulative, irrespective of PPI used.
Conclusion: Pantoprazole and rabeprazole do not significantly
affect liver function in patients with HCV-related liver cirrhosis.
Both PPIs are safe for treatment of patients with advanced liver
disease.
e64
F-45
F-46
TRANSIENT ELASTOGRAPHY IN PATIENTS WITH
CHRONIC AUTOIMMUNE HEPATITIS: IS A GOOD
TOOL IN EVALUATION OF HEPATIC FIBROSIS?
LITAF ROLE IN IN VIVO HEPATOCYTE
PROLIFERATION AND IN HEPATOCARCINOMA
CELLS
S. Onali 1,5 , S. Cappellini 1,5 , G. Serra 1,5 ,
T. Zolfino 2,5 , D. Murgia 3,5 , M.L. Ponti 3,5 ,
C. Balestrieri 1,5 , M. Conti 1,5 , O. Sorbello 4,5 ,
A. Civolani 4,5 , M. Casale 1,5 , S. Casu 1,5 ,
F. Figorilli 1,5 , M.C. Pasetto 1,5 , A. Gaspardini 1,5 ,
L. Secci 1,5 , C. Salustro 1,5 , R. Ganga 3,5 ,
M.R. Piras 2,5 , L. Demelia 4,5 , L. Chessa 1,5
N. Panera 1 , A. Crudele 1 , S. Ceccarelli 1 , D. Gnani 1 ,
C. De Stefanis 1 , V. Nobili 1 , A. Alisi 1
of Medical Sciences “M.Aresu”, University of Cagliari,
Italy
2 S.C. Gastroenterologia, AOB, Cagliari, Italy
3 S.C. Medicina Prima, AOB, Cagliari Italy
4 U.O. Gastroenterologia, AOU, Cagliari, Italy
5 Associazione Epatologi Sardi
Background and aims Liver stiffness measurement (LSM) by
transient elastography (TE, FibroScan) is a recently validated
method for noninvasive evaluation of fibrosis in chronic liver diseases, especially in patients with chronic hepatitis C.
The role of TE in evaluating liver fibrosis in chronic autoimmune
hepatitis (AIH) patients has not been yet extensively investigated.
Therefore, we aimed to assess the diagnostic performance of TE in
AIH patients by comparison with other nonivasive methods, such
as APRI and FIB-4 score.
Methods Sixty four patients with AIH were consecutevely
enrolled between January 2012 and August 2014. All patients were
in treatment with steroid and or other immunosoppessive drugs,
such as azathioprine. TE was performed in all patients and liver
stiffness value was compared with the histological stage (Metavir)
using the Spearman correlation. APRI and FIB-4 score were also
calculated. Patients who underwent to liver biopsy more than five
years before the LSM were excluded. Patient with a clinical diagnosis of cirrhosis and no signs of portal hypertension were also
included.
Results 22 patients were analysed: M/F:3/19; mean age 48
years; mean BMI 17 Kg/m2 ; mean AST 23UI/l (13-50); mean ALT
28 UI/l (13-100); most of the patients had mild-moderate fibrosis
(15/23). Mean liver stiffness value was 6,4 Kpa (3-18).
The Spearman test showed a significant correlation between
LSM and histological stage (r = 0,56; p = 0,007). Mean liver stiffness
value in cirrhotic patients was 10,6 Kpa vs 4,8 Kpa in non cirrhotic
patients (p < 0,001). A significant correlation was also observed
between FIB-4 score and histological stage (r = 0,62, p = 0,002),
although no differences were found between cirrhotic and non
cirrhotic value (p = 0,56).
Conclusions Transient elastography could be an effective
method to assess and monitor hepatic fibrosis in chronic autoimmune hepatitis, especially in patients with advanced fibrosis.
1 Liver Research Unit, Bambino Gesù Children
Hospital and IRCCS, Rome, Italy
Introduction: Liver regeneration and chronic liver injury
occurring during HCC development may result in compensatory
proliferation of differentiated hepatocytes. Several lines of evidence demonstrated that some pro-inflammatory cytokines (i.e.
TNF-a) and gut-derived products, such as lipopolysaccharide (LPS)
have been involved in the regulation of hepatocytes proliferation
and apoptosis.
Aim: The aim of the present study was to explore the role of
LITAF, an LPS-induced TNF-a transcription factor, in the regulation
of hepatocyte proliferation.
Materials and Methods: We established an in vivo model of
hepatocyte proliferation by 70% partial hepatectomy (PHx) in 24
male Wistar rats. We used this model to investigate, by real-time
PCR, Western Blot and immunohistochemistry analysis, the hepatic
expression of LITAF and of those proteins/genes involved in the
control of cell cycle and apoptosis. We further analysed the same
proteins/genes in HepG2 cells stably silenced with shRNA lentivirus
for LITAF.
Results: We found an up-regulation of LITAF mRNAs and protein expression after 1hr and 6 hrs from PHx. This LITAF increase
correlated with that of it transcriptional regulator, p53 and with
an up-regulation of PCNA, a well-known proliferation marker. No
apparent correlation of LITAF with the expression of other cell cycle
markers, such as cyclin D1 and cyclin B1, was found. These data suggest a potential role of LITAF in hepatocyte homeostasis regulation.
In fact, the proliferation rate was increased in stable LITAF-silenced
HepG2 cells with particular effect at early times of cells cycle, even
though the absence of LITAF unaffected the expression of cyclins
D1 and B1, cdk4 and cdk1 and TA, DTA and DN isoforms of p53 and
apoptosis rate.
Conclusions: In conclusions, this preliminary data indicates a
pivotal role of LITAF in regulating early times of first cell cycle and
hepatocytes proliferation both in in vivo liver regeneration model
and in transformed cells.
F-47
F-48
SARCOPENIA IS ASSOCIATED WITH A REDUCED
SURVIVAL IN PATIENTS WITH
HEPATOCARCINOMA UNDERGOING SORAFENIB
TREATMENT
ORAL BACTERIAL LOAD IN AUTOIMMUNE LIVER
DISEASES: POSSIBLE ROLE OR COINCIDENCE?
E. Gigante 1 , G. Antonelli 1 , P. Begini 1 ,
F. Carbonetti 2 , E. Iannicelli 2 , P. Marchetti 3 ,
L. Miglioresi 4 , A. Pellicelli 4 , G. Delle Fave 1 ,
M. Marignani 1
1 Digestive and Liver Diseases unit, St. Andrea
Hospital, Faculty of Medicine and Psychology,
University Sapienza’, Rome, Italy
2 Radiology unit, St. Andrea Hospital, Faculty of
Medicine and Psychology, University Sapienza’,
Rome, Italy
3 Oncology unit, St. Andrea Hospital, Faculty of
Medicine and Psychology, University Sapienza’,
Rome, Italy
4 Liver Unit, San Camillo-Forlanini Hospital, Rome,
Italy
Background and aim: Sarcopenia has been associated with
poor outcomes in patients with cirrhosis and solid tumors, and with
higher toxicity during chemotherapy. Hepatocarcinoma (HCC) is
diagnosed at advanced stage in up to 40% of all patients. At this
stage, the only approved treatment is Sorafenib. We analyzed the
influence of sarcopenia on survival of HCC patients treated with
Sorafenib.
Methods: We conducted a retrospective study on 41 patients
with advanced HCC treated with Sorafenib. Enrolled patients performed abdominal computed tomography (CT) 30 days within
treatment start, and data regarding survival, treatment toxicity,
anthropometric features, and laboratory findings were collected.
A transverse CT image from L3 was collected from each scan and
analyzed with SliceOmatic 5.0. The muscles’ cross-sectional area at
this level were selected and normalized for height, obtaining the
skeletal-muscle-index (SMI). Presence of sarcopenia was defined
by SMI ≤41 cm2/m2 for women, and ≤53cm2/m2 for men with
body mass index (BMI) ≥25, and ≤43 cm2/m2 for those with a BMI
<25.
Results: Patients were mainly males (80,5%). Sarcopenia was
present in 36,5% of patients (15/41). The two groups, with or
without sarcopenia were compared. Sex distribution and baselines
features were similar, no significant differences in albumin, INR,
BMI, Sodium, creatinine, bilirubin and MELD score were detected.
Child-Pugh Score was higher in the sarcopenic group [6 (5-8) vs 5
(5-8); p = 0,034]. Sarcopenic patients had reduced Overall Survival
79 (95% CI 44-114) vs 181(95% CI 140-223) weeks (p = 0,0013), as
well as smaller survival after treatment: 24 vs 54 weeks(figure 1).
Time on treatment of sarcopenic patients was 12,3 vs 22,9 weeks
(H.R. 1,83, p = 0,0464), but there were no differences in the cause of
interruption.
Conclusion: Sarcopenia is present in a third of advanced HCC,
and is associated with reduced OS and with reduction of time on
Sorafenib treatment.
e65
G. Orrù 1 , S. Onali 2 , C. Salustro 2 , S. Cappellini 2 ,
E. Cocco 1 , S. Fais 1 , M. Melis 1 , A. Gilardi 1 ,
B. Musu 1 , L. Secci 2 , G. Serra 2 , C. Balestrieri 2 ,
M. Conti 2 , M. Casale 2 , S. Casu 2 , F. Figorilli 2 ,
M.C. Pasetto 2 , R. Littera 3 , R. Scioscia 2 , L. Barca 2 ,
L. Chessa 2
1 DNA Sequencing Service, Department of Surgical
Sciences, University of Cagliari, Cagliari, Italy
of Medical Sciences “M.Aresu”, University of Cagliari,
Cagliari, Italy
3 Medical Genetics, Department of Medical Sciences
“M.Aresu”, University of Cagliari, Cagliari, Italy
Background and aims: Several factors seem be involved in the
pathogenesis of autoimmune liver disease (ALD), such as bacteria or bacterial components. Some studies highlighted the role of
an oral bacterium, P. gingivalis, in the pathogenesis of rheumatoid
arthritis and in the induction of NASH. We evaluated if there was a
correlation between oral bacteria and some features of ALD.
Methods: We analyzed 50 patients with ALD, 22 with autoimmune hepatitis (AIH) and 28 with primary biliary cirrhosis (PBC),
94% females, median age 61 years, median disease duration 7.5
years, 18% with cirrhosis, 66% responders to treatment, 40%
with concomitant autoimmune diseases, and 46 health controls
matched for sex and age. Patients and controls were subjected to
cytobrush of the tongue. Exclusion criteria were antibiotic use in
the previous week, mouthwash use on the day of inclusion. The
study was conducted with the local ethics committee approval
and informed consent was obtained from all subjects. Bacterial
DNA (BD) obtained from the tongue biofilm was quantified using
a computer RNA-DNA and expressed as ug/ml of sample.For each
analysis, three distinct biological replicas were done, and quantitative data were expressed as mean ± SD.
Results: BD amount was greater in ALD than in controls
(1834.10 vs. 881.75, p < 0.0001). There was no correlation between
BD amount and age, disease duration, histological findings and biochemical parameters, except for alkaline phosphatase (p = 0.05).
The presence of cirrhosis, response to therapy and the immunosuppressive drug use did not affect BD amount. Finally, patients with
concomitant autoimmune diseases had a greater BD amount than
the other (2053.54 vs. 1687.60) but without significance (p = 0.57).
Conclusion: Our study showed an unspecified role of oral bacteria in patients with autoimmune liver diseases. Further analysis
will be needed to assess if there are differences in the microbial
population.
e66
F-49
F-50
EFFICACY OF SORAFENIB IN PATIENTS WITH
INTERMEDIATE-STAGE HEPATOCELLULAR
CARCINOMA: RESULTS FROM THE ITA.LI.CA.
DATABASE
CORRELATION BETWEEN NK CELLS AND
RESPONSE TO TRIPLE HCV-THERAPY WITH
BOC/TVR
R. Sacco, V. Mismas, A. Romano, M. Bertini,
M. Bertoni, G. Federici, G. Parisi, S. Metrangolo,
E. Tumino, G. Bresci
Department of Gastroenterology, Pisa University
Hospital, Pisa, Italy, for ITA.LI.CA group
S. Marenco 1 , G. Pieri 1 , F. Mazza 1 , M. Brunacci 1 ,
S. Labanca 1 , L. Bruzzone 1 , F. Valentina 1 ,
F. Bozzano 2 , F. Marras 3 , A. De Maria 3 ,
V. Savarino 1 , A. Picciotto 1
1
U.O.s. Diagnosi e Terapia delle Epatopatie, IRCCS,
San Martino IST, Genova, Italy
2 Istituto G.Gaslini, Genova, Italy
3 Centro di Eccellenza per la Ricerca Biomedica,
Universita di Genova, Genova, Italy
Introduction Sorafenib represents the elective treatment for
patients affected from advanced-stage (BCLC-C according to
the Barcelona Clinic of Liver Cancer classification) HCC; this
molecule is also recommended in patients with intermediatestage (BCLC-B) disease who have failed or are not eligible to
transareterial chemoembolization (TACE). However, further information on the use of sorafenib in patients with BCLC-C HCC is
warranted.
Aims This study analyses the efficacy of sorafenib in patients
with BCLC-B HCC, with respect to those with BCLC-C disease, in the
Nation-wide Italian database ITA.LI.CA.
Patients and methods The ITA.LI.CA. database contains data of
5428 HCC patients treated at 18 Italian Centers. All patients with
either BCLC stage B or C and who received sorafenib were considered. Overall survival (OS), time to progression (TTP), and disease
control rate (DCR) were evaluated. Safety considerations were also
performed.
Results A total of 243 patients were included. Of these, 61
were in BCLC-B stage (29 received ≥1 TACE prior to sorafenib)
and 182 in BCLC-C. In the overall population, median TTP was
8 months (95%CI 6-9), median OS was 13 months (95%CI 1015.5), and DCR was 28.3% (complete response, CR = 1.6%; partial
response, PR = 8.9%; stable disease, SD = 17.8%). Patients in BCLCB stage had a median TTP of 10 months (95%CI 6-14), a median
OS of 19 months (95%CI 12.5-26.5), and a DCR of 39.1% (CR = 2.4%;
PR = 9.8%; SD = 26.8%), whereas patients in BCLC-C stage had
a median TTP of 7 months (95%CI 6-8), a median OS of 11
months (95%CI 10-15.5), and a DCR of 25.3% (CR = 1.3%; PR = 8.7%;
SD = 15.3%). The safety profile of sorafenib was similar in the two
sub-populations.
Conclusions These “field-practice” findings suggest that
the administration of sorafenib in BCLC-B patients with HCC
may be effective and is not associated with any new safety
warning.
Background and aims. In our previous study on 28 naïve
genotype-1 patients, we showed that the CD56bright CD16+/− cell
populations and the expression of both activating and inhibitory
NK-cell receptors were reduced at baseline in patients with sustained virological response (SVR), after dual therapy with pegylated
interferon-alpha and ribavirin (IFN/R). The aim of thos study is to
verify the correlation between subset cell populations or activating and inhibitory NK-cell receptors and response to triple therapy
with boceprevir (BOC) or telaprevir (TVR).
Methods. We prospectively enrolled 33 patients between Jan
2012 and Jan 2013, candidate to triple therapy with BOC/TVR.
29/33 patients were male, 31/33 non-responder (NR) to IFN/R,
21/33 had cirrhosis. We evaluated NK cells populations, activating
and inhibitory receptors before starting triple therapy (baseline).
Statistycal analysis was performed with Mann–Whitney U-test. NK
cells subset populations were analyzed by cytofluorometric assay
(Becton Dickinson, Mountain View, CA, USA). We used specific
mouse anti-human mAb to determine the receptors.
Results. 26 patients were treated with TVR and 7 with BOC.
17/33 patients obteined SVR, while 16 did not respond (NR). We
found no differences in the expression of both activating (NKp30,
NKp46, DNAM-1) and inhibitory receptors (CD85j) or the subset
cell populations (CD56dull CD16+ , CD56bright CD16+/− ) between SVR
and NR patients (p > 0,08).
Conclusions. As previously demonstrated, differences in the
expression of activating and inhibitory NK cell receptors contribute
to the differential response to dual treatment. This evidence was
not confirmed in patient treated with triple therapy with BOC/TVR.
Therefore, the evaluation of NK cells characteristics at baseline is
useless in predicting virological response to triple therapy with
BOC/TVR.
Digestive and Liver Disease 47S (2015) e67
A.I.S.F. 2015: Abstracts evaluation procedure
Thanks to experts evaluating all the abstracts according to predetermined Clinical and Experimental categories.
The experts for the 2015 Annual Meeting are listed below:
Category A. “HEPATITIS C”
Clinical/Experimental
P.A. Aghemo, Milan - P. Andreone, Bologna - D. Bitetto,
Bergamo - S. Bruno, Milan - V. Di Marco,
Palermo - A. Mangia, San Giovanni Rotondo (FG) G. Missale, Parma - F. Morisco, Naples - P. Pontisso,
Padua - G. Taliani, Rome - A.L. Zignego, Florence
Category B. “HEPATITIS B & DELTA”
B. Coco, Turin - V. Di Marco, Palermo - C. Ferrari,
Parma - M. Iavarone, Milan - A. Marzano, Turin - F.P. Russo,
Padua - G. Taliani, Rome - M. Viganò, Milan
Category C.
Category F.
“AUTOIMMUNE HEPATITIS & BILIARY DISEASE”
V. Cardinale, Rome - A. Floreani, Padua - L. Muratori,
Bologna - C. Rigamonti, Novara - F. Rosina, Turin - C. Spirli,
New Haven, CT (USA)
“METABOLIC LIVER DISEASE (Emocromatosi,
Wilson, etc.)”
A. Cappon, Padua - S. Fargion, Milan - A. Lleo, Rozzano
(MI) - F. Marra, Florence - G. Svegliati-Baroni, Ancona
1590-8658/
Category H. “PORTAL HYPERTENSION”
A. Berzigotti, Barcelona (Spain) - F. Campagna,
Padua - P. Caraceni, Bologna - A. Ciaccio, Monza - F. Dell’Era,
Milan - V. La Mura, Milan - F. Schepis, Modena
Category I.
“CIRRHOSIS & ITS COMPLICATIONS”
A. Berzigotti, Barcelona (Spain) - F. Campagna,
Padua - P. Caraceni, Bologna - A. Ciaccio,
Monza – F. Dell’Era, Milan - V. La Mura, Milan - F. Schepis,
Modena
Category L.
“HEPATOCELLULAR CARCINOMA”
S. Bhoori, Milan - F. Farinati, Padua - E.G. Giannini,
Genoa - L. Gramantieri, Bologna - E. Gringeri,
Padua - M. Iavarone, Milan - G. Missale, Parma- F. Piscaglia,
Bologna - E. Villa, Modena
“ALCOHOLIC LIVER DISEASE”
G. Germani, Padua - C. Loguercio, Naples - C. Puoti,
Marino (Rome)
Category D. “NON-ALCOHOLIC FATTY LIVER DISEASE”
P. Dongiovanni, Milan - L. Miele, Rome - S. Petta,
Palermo - E. Vanni, Turin
Category E.
Category G. “FIBROSIS”
M. Fraquelli, Milan - F. Marra, Florence – M. Parola,
Turin - G. Svegliati-Baroni, Ancona - F. Vizzutti, Florence
Category M. “HEPATOBILIARY SURGERY & LIVER
TRANSPLANTATION”
M. Angelico, Rome - L. Baiocchi, Rome - P. Burra,
Padua - M.F. Donato, Milan - S. Fagiuoli, Bergamo
Category N. “ACUTE LIVER INJURY & HEPATOTOXICITY”
M. Fraquelli, Milan - G. Germani, Padua – A. Gasbarrini,
Rome - F. Giannone, Modena

Italian Association for the Study of the Liver – Annual Meeting

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