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Hepatitis C VirusTowards 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 MondorUniversité Paris-EstCréteil, France

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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- and ribavirin– Pegylated IFN-, ribavirin and a protease inhibitor (genotype 1)

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A Truly Translational Setting

Clinical research unitClinical trials

Cohort studies

Clinical virology labNational Reference Center

for Viral Hepatitis B, C and D

Basic research lab INSERM U955

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Clinical Group

Christophe HézodeAnne Varaut

Murielle FrançoisMarie Payet

Isabelle RivièreOlivier Teston

Isaac Ruiz Ariane Mallat

Clinical Virology Group and National Reference Center

For Viral Hepatitis B, C and D

Stéphane ChevaliezDominique Challine

Magali Bouvier-Alias,Jérémie CorneilleAlexandre SoulierNikolaos GatselisFrançoise DarthuyCharlène Prénom

Drug screening and ResistanceINSERM U955

Abdelhakim Ahmed-BelkacemPaul Ben Sadoun

Coralie PallierChristophe Rodriguez

Nazim AhnouEva HernandezRozenn Brillet

Liver CarcinogenesisINSERM U955

Hervé LeratMartin Higgs

Philippe ChouteauNicole Defer

Thomas DecaensMuhamad Ahmad Maqbool

Alexandre FlorimondMohamed Imache

Aurore Gaudin

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A Multidisciplinary Team

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HCV Clinical Trials

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2005 2006 2007 2008 2009 2010

Phase IPhase IIPhase IIIPhase IV

Nu

mb

er o

f cl

inic

al t

rial

s st

arte

d

2005-2010: 586 patients included

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Viral genome

extraction from serum

RT PCR amplification

emPCR Pyrosequencing

PyroMute® PyroDyn®

% of each mutations

Analysis

Data collection

PyroClass®

PyroClass©. PyroMute©, PyroDyn©, PyroLink© are protected under IDDN

Modeling of population growth

Sequence quality filters

PyroLink®

Linkages

Classificationof generated sequences

(Rodriguez C. et al., AASLD 2010)

New Virological ToolsNext-Generation Sequencing

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HCV Resistance by UDPS

(Chevaliez S. et al., EASL 2011)H

CV R

NA

(Log

10 IU

/mL)

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2

4

6

8

Days of treatment

% o

f mut

ation

s in

the

who

le q

uasi

spec

ies

% o

f var

iant

s in

the

quas

ispe

cies

Days of therapy

HCV

RN

A (L

og10

IU/m

L)

Viral populations

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Models for the Study of HCV-Induced Hepatocarcinogenesis

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Liver tissues fromHCV-infected patientswith or without HCCs

HCV-infected hepatoma cell lines harboring HCV replicons or infectious

HCV strains

HCV transgenic mousemodel (FL-N/35) expressing

the full HCV ORF*

(*Lerat H, et al., Gastroenterology 2002;122::352-65)

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HCV-Induced Hepatocarcinogenesis

HCV proteinexpression

CH3 CH3

Méthylation

Promoteurde Gadd45

ROS production

DNA damage

-catenin

Promoteurde c-Myc

c-Myc

Impaired G2-M arrestDefective DNA damage repair

Gadd45

Genomic instabilityDefective G1-S arrest

Impaired DNA damage repair

(Higgs M, et al., Cancer Res 2010;70:4901-11)

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HCV Drug Development

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

(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

IC50 (M) cyclophylin A

EC50 (M) HCV-1b 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

(Ahmed-Belkacem A, et al., AASLD 2010)

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Perspectives12

Vaccine ResearchInstitute (VRI)