Christine MENETRIER-CAUX

3
PI
Plateforme
d’Innovation
en Immuno-monitoring
et Immunothérapie
LYMPHOS’1 Project
C A N C É RO P Ô L E LYO N A U V E RG N E R H Ô N E - A L P ES
A C C É L É R A T E U R D ’ A V A N C É ESS
OUS L'ÉGIDE DE LA FONDATION BULLUKIAN
LYMPHOS’1 Project Identity Card
PI3
Plateforme
d’Innovation
en Immuno-monitoring
et Immunothérapie
2 partners : Centre Léon Bérard / Centre de Recherche en Cancérologie de Lyon
(CRCL) & ImmunID Technologies
2 localizations : Lyon / Grenoble
Project Duration: 4 years (2008-2012)
Global Budget : 853 k€ (50% for the academic partner financed by CLARA/Rhône)
Proof of concept: Demonstrate the clinical interest to detect
systemic immune dysfunctions in metastatic breast carcinoma
Objective: Validate a prognostic test to characterize at risk
patients that will benefit from innovative therapeutics
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PI3
SCIENTIFIC BACKGROUND
Plateforme
d’Innovation
en Immuno-monitoring
et Immunothérapie
• 20-30% patients with metastatic solid tumors show a lymphopenia associated
with complications:
• Febrile neutropenia (Blay JY et al. JCO 1996 92:405-10)
• Grade IV anemia and thrombocytopenia (Ray-Coquard I. et al. JCO 1999 17:2840-6)
• Increased risk of Early Death (48%) (Ray-Coquard I. et al. Br J Cancer 2001 85:816-22)
• Lymphopenia detected before chemotherapy is an independent pronostic
factor for overall survival in metastatic solid tumors (Borg C et al. Cancer 2004
101:2675-80; Ray-Coquard I. et al Cancer Res 2009 69:5383-91)
Metastatic Breast Carcinoma (n= 290)
Overall survival (%)
Overall survival (%)
Log Rank test =21.5, p<0.01
100
80
lymphocytes (/µL)
>1000/µL
<1000/µL
60
40
20
0
Soft tissue Sarcoma (n=193)
0
20
40
60
Months
DATE
80
100
100
90
80
70
60
50
40
30
20
10
0
Log Rank test: p=0.0057
lymphocytes (/µL)
>1000/µL
<1000/µL
0
1
2
3
NOM DE LA PRÉSENTATION (INSERTION - EN-TÊTE ET PIED DE PAGE)
7
4
5
6
8
Years
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
3
ImmunID Technology to assess TCR diversity
Identification of V-J rearrangements at the genomic level
Stability of samples
Avoid bias due to transcriptional regulation (1 cell = 1 rearrangement)
PCR multi-N-plex
(V-J)
Example : TRAD Locus
Flow
Cytometry
(V)
Spectratyping &
Sequencing
(CDR3)
V genes
J genes
AC
gDNA
A
C
Vn Ji
V-Ja
Env. 10Kb
Rearrangement
Splicing
Vn - Ji A
C
Hz
Combinatorial diversity : V x (D) x J
Jonction diversity: CDR3
V
Matching diversity : TCRa x TCRb
Somatic hypermutations
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
J
TCR repertoire diversity is altered in metastatic breast cancer
patients and has an added value to lymphopenia status
Divpenia<1st tertile
Lymphopenic Patients
Non Lymphopenic
Patients
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Comparaison of the distribution of patients according
to the NDL score
Metastatic Breast
Cancer Patients
Healthy donors
NDL4
NDL3
NDL4
NDL1
NDL2
NDL1
NDL2
Lymphocyte absolute
Numbe (Giga/L)
NDL3
combinatorial diversity hTRB VJ (%)
Does NDL classification help to discriminate patients prognosis ?
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NDL-1 score characterizes a population of patients
with increased risk of early death
Manuel M, et al. Onco-Immunology 1:433-42, 2012.
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
NDL Score is an Independent factor of poor prognosis
for metastatic breast cancer patients
Multivariate analysis integrating all valuable clinical parameters (Triple negative, PS, Anemia,
number of metastatic sites, menopause, LDH levels)
Manuel M, et al. Onco-Immunology 1(4):433-42, 2012.
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In depth analysis of the impact on
patient survival of different
lymphocyte subset lymphopenia
DATE
NOM DE LA PRÉSENTATION (INSERTION - EN-TÊTE ET PIED DE PAGE)
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
9
No impact of Defect in B cells and NK cells numbers
on survival in contrast to CD8+ T cells lymphopenia
CD56+ Lymphopenia
CD8+ Lymphopenia
% patients alive
CD19+ Lymphopenia
Follow-up (months)
Tredan
et al.
Eur
J Cancer
2013 in press
SOUSO,
L'ÉGIDE
DE LA
FONDATION
BULLUKIAN
CD4+ lymphopenia impacts strongly patients’ survival in
first relapse but is less determinant at later stages
1st relapse
CD4+< 0.2 (n=16)
0.2 ≤CD4+ > 0.45 (n=39)
CD4+≥ 0.45 (n=48)
1,0
0,8
% survvial
>2nd relapse
0,8
0,6
0,6
0,4
0,4
0,2
0,2
Log rank p-value=10-4
0,0
0
10
20
30
40
50
CD4+< 0.2 (n=25)
0.2 ≤CD4+ > 0.45 (n=46)
CD4+≥0.45 (n=30)
1,0
0,0
Log rank p-value=0.07
0
10
20
30
40
50
60
Follow-up (months) Follow-up (months)
SOUSO,
L'ÉGIDE
DE Eur
LA FONDATION
BULLUKIAN
Tredan
et al.
J Cancer
2013 in press
CD4+ lymphopenia an independent factor of poor prognosis in
metastatic breast cancer patients
Multivariate analysis integrating all valuable clinical parameters (Triple negative, PS,
anemia, number of metastatic sites, Menopause, neutropenia) whatever the line of
treatment in metastatic setting
1st Relapse
>2nd Relapse
HR
CI 95%
P-value
Menopause (Yes Vs No)
1.878
[1.080 - 3.266]
0.026
Neutropenia (<7.5 Vs ≥7.5) (Giga/L)
4.201
[2.202 - 8]
0.000
CD4+ Lymphopenia (<0.45 Vs ≥0.45) (Giga/L)
2.521
[1.543 - 4.490]
0.000
HR
CI 95%
P-value
PS (0/1 Vs >1)
1.871
[1.079 - 3.245]
0.026
LDH (<600 Vs ≥600) (UI/L)
3.330
[1.877 – 5.908]
0.000
CD4+ Lymphopenia (<0.45 Vs ≥0.45) (Giga/L)
2.137
[1.204 - 3.792]
0.009
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
BETTER UNDERSTAND TO REVERSE
…. CAMEL Project (PI: O. Tredan)
Patients with metastatic breast cancer leading to
CD4+ T cell lymphopenia have poor outcome
Trédan Olivier1, Manuel Manuarii2,3,4,5,7, Clapisson Gilles2, Bachelot Thomas1,3,4,5, Chabaud
Sylvie6, Bardin-dit-Courageot Christine2, Rigal Chantal2, Biota Cathy 2,3,4,5, Bajard Agathe6,
Pasqual Nicolas7, Blay Jean-Yves1,2,3,4,5, Caux Christophe2,3,4,5 and Ménétrier-Caux
Christine2,3,4,5
BUT WHY ???
TUMOR
ALTERATIONS ?
HOST CONSTITUTIONAL
CHARACTERISTICS?
COMBINATION OF ALTERATIONS ?
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
BETTER UNDERSTAND ….
Does Lympho(-div)penia result from TUMOR ALTERATIONS ?
Analysis on Tumor DNA from FFPE samples
• Driver mutations
BRAF, PI3KCA, ALK, FGFR1, EGFR…
• Mutations in “immune-related “ genes
Tumor
Area
Evaluation of gene mutations
HTS Sequencing
NGS-Ion Torrent
Evaluation of gene
amplification/deletion
CGH Array
BETTER UNDERSTAND…..
Does Lympho(-div)penia result from TUMOR ALTERATIONS ?
Preliminary data on 3 patients from LYMPHOS1
Pt 1
Mutated Genes
BRAF
CRAF
MET
PIK3CA
PIK3R1
mTOR
ERBB2
EGFR
PTEN
PDGFA
PDGFB
PDGFRA
PDGFRB
cKIT
ALK
RB1
RET
…
Pt 2
FGFR-1
RB1
Gene
DDR2
EGFR
ERBB2
PDGFA
PIK3R1
RB1
RYK
SDHC
TP53
AURKB
FGFR-1
PIK3CB
Localisation
1q23.3
7p12
7q12
7p22
5q13.1
13q14.2
3q22
1q23.3
17p13.1
17p13.1
8p12
3q22.3
Type of
Aberration Druggable
gain
Yes
loss
Yes
loss
Yes
loss
Yes
loss
Yes
loss
No
loss
No
gain
No
loss
No
loss
No
amplification
Yes
loss
No
TP53
FGFR-1 FISH
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
BETTER UNDERSTAND…..
Does Lympho(-div)penia result from HOST CONSTITUTIONAL
CHARACTERISTICS ?
Analysis on frozen PBMC
Evaluate immune Polymorphisms that could alter immune homeostasis
Apetoh
L.L'et
alDEImmunol
2007 220:47-59
SOUS
ÉGIDE
LA FONDATION Rev
BULLUKIAN
BETTER UNDERSTAND…… TO REVERSE
- Tumor alterations
- Host polymorphisms/Immunity
• Drug to compensate polymorphism:
• Chloroquine for TLR4, Activation of other TLR signalling
• Stimulation of pathway downstream receptor polymorphism (IL-7R)
• Use of drugs targeting immune-suppressive molecules (PDL1, CTLA4)
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
OUR PERCEPTION OF
CONTRIBUTION OF “POC” PROGRAM
IMMUNID
• Technological improvement of the
hTRB test
• Access to scientific and clinical
expertise for strategic direction
• Validation of a clinical application in
metastatic breast cancer for the
prognostic test developed by
ImmunID
CLB/CRCL
• Access to innovative technologies
• Validation of concepts
• Co-development of new use for the
benefit of patients
• Validation of robust immunomonitoring tools (bio-marker) and
initiation of new clinical trials
• Creation of value through the filing
of a joint patent
• Generation of shared intellectual
property
• Publication of two scientific articles
involved in the awareness of the
society
• Publication of two scientific articles
• Maintain and create jobs
SOUS L'ÉGIDE DE LA FONDATION BULLUKIAN
PI3 Platform & CRCL
Christophe CAUX & Jean Yves BLAY
Manuarii MANUEL
Julien FAGET
Christine MENETRIER-CAUX
Gilles CLAPISSON
Christine BARDIN
Chantal RIGAL
ImmunID
Nicolas PASQUAL
Manuarii MANUEL
Tioka RABEONY
Audrey GRIVES
Jean-François MOURET
Audrey NONDE
Sébastien WEISBUSCH
CRB
Gilles CLAPISSON
Séverine MARTINEZ
Laetitia ESCALES
Amélie ALLAUZE
CLB Physicians
Thomas BACHELOT
Olivier TREDAN
Jean-Yves BLAY
Aude DURET
Jean Pierre GUASTALLA
Sana -Intidhar LABIDI
Maurice PEROL
Isabelle RAY COQUARD
Paul REBATTU
BEC
Nathalie GAUTHIER
Arlette DUMORTIER/Christelle FLORENT
UBET
David PEROL
Sylvie CHABAUD
Séverine DENIZOT-GUILLEMAUT
Séverine METZGER
Leila BEN ABDESSELEM
Nathalie GIRERD-CHAMBAZ
UGAF
Zora ABDELBOST
Liliana VIEIRA
Marina ROUSSEAU-TSANGARIS
UBGDI
Thérèse GARGI
Hôpital Lyon Sud
Dr Pierre-Jean SOUQUET
Patients
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