Transcript 2-4-jean-michelpawlotsky-110610091609-phpapp02

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Hepatitis C Virus
Towards eradication of an
oncogenic viral agent
Prof. Jean-Michel Pawlotsky
National Reference Center for
Viral Hepatitis B, C and Delta,
Department of Virology
& INSERM U955
Hôpital Henri Mondor
Université Paris-Est
CEA
CHRU
CNRS
Créteil, France
CEA
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CNRS
CPU
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Hepatitis C Virus Infection
• 120-130 million individuals chronically infected worldwide
• 1st cause of chronic liver disease and 1st cause of
hepatocellular carcinoma (HCC, primary liver cancer) in
the industrialized world
• Mortality rate: >300,000/year
• Curable by therapy
– Pegylated IFN-a and ribavirin
– Pegylated IFN-a, ribavirin and a protease inhibitor (genotype 1)
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A Truly Translational Setting
Clinical research unit
Clinical trials
Cohort studies
Clinical virology lab
National Reference Center
for Viral Hepatitis B, C and D
Basic research lab
INSERM U955
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A Multidisciplinary Team
Clinical Group
Drug screening and Resistance
INSERM U955
Christophe Hézode
Anne Varaut
Murielle François
Marie Payet
Isabelle Rivière
Olivier Teston
Isaac Ruiz
Ariane Mallat
Abdelhakim Ahmed-Belkacem
Paul Ben Sadoun
Coralie Pallier
Christophe Rodriguez
Nazim Ahnou
Eva Hernandez
Rozenn Brillet
Clinical Virology Group and
National Reference Center
For Viral Hepatitis B, C and D
Liver Carcinogenesis
INSERM U955
Hervé Lerat
Martin Higgs
Philippe Chouteau
Nicole Defer
Thomas Decaens
Muhamad Ahmad Maqbool
Alexandre Florimond
Mohamed Imache
Aurore Gaudin
Stéphane Chevaliez
Dominique Challine
Magali Bouvier-Alias,
Jérémie Corneille
Alexandre Soulier
Nikolaos Gatselis
Françoise Darthuy
Charlène Prénom
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HCV Clinical Trials
Number of clinical trials started
8
7
2005-2010:
586 patients included
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Phase I
Phase II
Phase III
Phase IV
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4
3
2
1
0
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2005
2006
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2007
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2008 INSERM 2009
2010
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New Virological Tools
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Next-Generation Sequencing
Data collection
RT
PCR amplification
emPCR
Pyrosequencing
Viral genome
extraction from serum
Analysis
% of each
mutations
©. PyroMute
©, PyroDyn©
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PyroClass
, PyroLink© are
under IDDNINRIA
(Rodriguez C. et al., AASLD 2010)
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HCV Resistance by UDPS
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H28Q+R155K
H28Q+R155K+S54T+Y52C
80%
H28Q+R155K+S54T+Y52C+V36M+H57
L+P96H
V36M+R155K+H57L
60%
R155K
8
40%
20%
0%
6
0
4
29
57
2
85
182
Days
Daysofoftreatment
therapy
(Log
RNA
HCV
(Log
RNA
HCV
10 IU/mL)
10 IU/mL)
quasispecies
whole
in the
% of mutations
quasispecies
in the
% of variants
100%
0
595
686
Viral populations
903
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(Chevaliez S. et al., EASL 2011)
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Models for the Study of HCV-Induced
Hepatocarcinogenesis
Liver tissues from
HCV-infected patients
with or without HCCs
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HCV-infected hepatoma
cell lines harboring HCV
replicons or infectious
HCV strains
CPU
(*Lerat H, et al., Gastroenterology 2002;122::352-65)
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HCV transgenic mouse
model (FL-N/35) expressing
the full HCV ORF*
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HCV-Induced Hepatocarcinogenesis
HCV protein
expression
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Promoteur
de Gadd45b
CH3
b-catenin
CH3
Méthylation
ROS production
Promoteur
de c-Myc
Gadd45b
c-Myc
DNA damage
Defective G1-S arrest
Impaired DNA
damage repair
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Impaired G2-M arrest
Defective DNA
damage repair
Genomic instability
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(Higgs M, et al., Cancer Res 2010;70:4901-11)
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HCV Drug Development
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Binding modes in HCV RNA-dependent RNA polymerase Thumb Pocket I of aurone inhibitors of HCV replication. (A) Compound 1 is involved in five
hydrogen bonds: two with Arginine 503 (1.9 Å and 2.5 Å), one with Glycine 493 (1.8 Å) and two with Leucine 392 (2.0 Å and 2.0 Å). (B) Compound 51 is
involved in three hydrogen bonds: two with Arginine 503 (1.8 Å and 2.0 Å) and one with Glycine 493 (2.2 Å). The pictures were built using Pymol
software
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(Haudecoeur R, Ahmed-Belkacem A, et al.,
J Med Chem 2011; in press)
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HCV Drug Development
Fragment-Based Drug Design
(FBDD) approach for small
molecule cyclophylin inhibitors
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(Ahmed-Belkacem A, et al., AASLD 2010)
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IC50 (mM)
cyclophylin A
EC50 (mM) HCV1b replicon
compound A
0.4±0.1
4.5±0.8
compound B
3.3±1.4
1.4±0.2
compound C
0.8±1.2
1.0±0.3
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Perspectives
Vaccine Research
Institute (VRI)
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