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1 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 CHRU CNRS CPU INRA INRIA INSERM INSTITUT PASTEUR IRD CPU INRA INRIA INSERM INSTITUT PASTEUR IRD 1 2 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) CEA CHRU CNRS CPU INRA INRIA INSERM INSTITUT PASTEUR IRD 3 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 CEA CHRU CNRS CPU INRA INRIA INSERM INSTITUT PASTEUR IRD 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 CEA CHRU CNRS CPU INRA INRIA INSERM INSTITUT PASTEUR IRD 4 5 HCV Clinical Trials Number of clinical trials started 8 7 2005-2010: 586 patients included 6 Phase I Phase II Phase III Phase IV 5 4 3 2 1 0 CEA CHRU 2005 2006 CNRS CPU INRA 2007 INSTITUT PASTEUR 2008 INSERM 2009 2010 INRIA IRD New Virological Tools 6 Next-Generation Sequencing Data collection RT PCR amplification emPCR Pyrosequencing Viral genome extraction from serum Analysis % of each mutations ©. PyroMute ©, PyroDyn© CEA CHRU CNRS CPUprotectedINRA PyroClass , PyroLink© are under IDDNINRIA (Rodriguez C. et al., AASLD 2010) INSERM INSTITUT PASTEUR IRD HCV Resistance by UDPS 7 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 CEA CHRU (Chevaliez S. et al., EASL 2011) CNRS CPU INRA INRIA INSERM INSTITUT PASTEUR IRD 8 Models for the Study of HCV-Induced Hepatocarcinogenesis Liver tissues from HCV-infected patients with or without HCCs CEA CHRU CNRS HCV-infected hepatoma cell lines harboring HCV replicons or infectious HCV strains CPU (*Lerat H, et al., Gastroenterology 2002;122::352-65) INRA INRIA HCV transgenic mouse model (FL-N/35) expressing the full HCV ORF* INSERM INSTITUT PASTEUR IRD HCV-Induced Hepatocarcinogenesis HCV protein expression 9 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 CEA CHRU Impaired G2-M arrest Defective DNA damage repair Genomic instability CNRS CPU (Higgs M, et al., Cancer Res 2010;70:4901-11) INRA INRIA INSERM INSTITUT PASTEUR IRD HCV Drug Development 10 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 CEA CHRU CNRS CPU INRA INRIA INSERM INSTITUT PASTEUR IRD (Haudecoeur R, Ahmed-Belkacem A, et al., J Med Chem 2011; in press) 11 HCV Drug Development Fragment-Based Drug Design (FBDD) approach for small molecule cyclophylin inhibitors CEA CHRU CNRS (Ahmed-Belkacem A, et al., AASLD 2010) CPU 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 INRA INRIA INSERM INSTITUT PASTEUR IRD 12 Perspectives Vaccine Research Institute (VRI) CEA CHRU CNRS CPU INRA INRIA INSERM INSTITUT PASTEUR IRD
