2.4 jean-michel pawlotsky
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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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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)
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