60 SVR12 RATES AND SAFETY OF TRIPLE THERAPY INCLUDING TELAPREVIR OR BOCEPREVIR IN 221 CIRRHOTIC NON RESPONDERS TREATED IN THE FRENCH EARLY ACCESS PROGRAM (ANRS CO20-CUPIC)

ORAL PRESENTATIONS several host cell factors, including CD81 and claudin1. We recently identified a role of epidermal growth factor receptor (EGFR) in HCV entry by regulating CD81-claudin1 co-receptor interaction, suggesting that EGFR-signaling plays a role in virus entry. To identify the molecular mechanisms of EGFR-signaling pathway(s) required for HCV entry, we aimed to uncover the molecular link between EGFR-signaling and the regulation of CD81-claudin1 coreceptor association. Methods: We combined RNAi screening in Huh7.5.1 cells, small molecule inhibition into primary human hepatocytes (PHH), phosphoarray analysis in PHH and liver biopsies and advanced imaging studies including fluorescence recovery after photobleaching (FRAP) and fluorescence resonance energy transfer (FRET). Results: RNAi screening comprising cellular signaling pathways demonstrated that silencing the GTPase HRas markedly inhibited HCV entry. Moreover, a trans-dominant active HRas mutant increased HCV entry and rescued the entry defect following EGFR inhibition, suggesting that HRas is a molecular switch regulating EGFR-mediated HCV entry. Indeed, FRET analyses demonstrated that similar to EGFR, silencing HRas expression reduced CD81claudin1 association. Moreover, differential proteomic analyses (SILAC) identified HRas specific association with a complex containing CD81. FRAP assays demonstrated that EGFR/HRas functions modulate lateral diffusion and membrane trafficking of CD81, allowing to form the host co-receptor complex. Furthermore, proteomic analyses identified novel signaling effectors within tetraspanin-enriched microdomains as candidates mediating this essential step of the HCV life cycle. Conclusion: Taken together, our results support a model where HRas/CD81 complexes provide a functional link between the EGFR/HRas pathway and CD81-claudin1 association that are prerequisites of HCV entry and indicate a crucial role of EGFR/HRas signaling for CD81 trafficking. These data increase our understanding of the molecular mechanisms underlying EGFR-mediated HCV entry and open new perspectives for the development of antivirals targeting signaling pathways. 59 LYMPHOTOXIN beta RECEPTOR ACTIVATION LEADS TO DEGRADATION OF HBV cccDNA FROM INFECTED HEPATOCYTES J. Lucifora, F. Reisinger, Y. Xia, R. Bester, K. Zhang, M. Sprinzl, U. Protzer, M. Heikenwalder. Institute of Virology, Technische Universit¨ at M¨ unchen/Helmholtz Zentrum M¨ unchen, Munich, Germany E-mail: [email protected] Background and Aims: Despite availability of efficient therapies against hepatitis B virus (HBV), long-term persistence of cccDNA requires life-long treatments that may cause side effects. The discovery of compounds directly targeting the cccDNA is thus one of the major challenges to cure chronic HBV infections. Here we aimed at testing the effect of lymphtoxin beta receptor (LTbR) agonization on HBV infection and replication. Method: Cell culture models including HBV-infected HepaRG cells and primary human hepatocytes as well as HBV-transgenic mice were used to test the effect of antibodies agonizing human LTbR (BS1) or mouse LTbR. Results: We observed a strong and dose-dependent anti-HBV effect by activation of the LTbR both in vitro and in vivo. All HBV replication markers were decreased upon treatments including cccDNA in cells where HBV infection was already established. The non-cytopathic degradation of cccDNA was found to be dependent on NF-kB activation through RelA phosphorylation but was independent of non-canonical NF-kB signaling or production of type I interferon. LTbR-activation resulted in overexpression of APOBEC3B resulting in hypermutations of cccDNA and subsequent specific degradation of cccDNA. As a consequence, and in contrast to

nucleos(t)ide analogues currently used for the treatment of chronic HBV infection, no rebound of HBV replication was observed after stop of treatment. Conclusions: We here describe an effective, long lasting and noncytopathic mechanism of effective HBV-cccDNA depletion – forming the basis for a novel alternative therapeutic approach to cure chronic HBV infection. 60 SVR12 RATES AND SAFETY OF TRIPLE THERAPY INCLUDING TELAPREVIR OR BOCEPREVIR IN 221 CIRRHOTIC NON RESPONDERS TREATED IN THE FRENCH EARLY ACCESS PROGRAM (ANRS CO20-CUPIC) H. Fontaine1 , C. Hezode2 , C. Dorival3 , D. Larrey4 , F. Zoulim5 , V. De Ledinghen6 , V. Canva7 , L. Alric8 , M. Bourliere ` 9 , S. Pol10 , T. Poynard11 , G. Riachi12 , P.-H. Bernard13 , J.-J. Raabe14 , J. Gournay15 , 16 S. Metivier ´ , J.-M. Pawlotsky17 , D. Samuel18 , Y. Barthe3 , F. Carrat3 , J.-P. Bronowicki19 , ANRS CO 20 CUPIC Study Group. 1 Hepatology Unit, Cochin Hospital AP-HP, Paris, 2 Hepatology Unit, Henri Mondor Hospital, Cr´eteil, 3 UMR-S 707, INSERM, Paris, 4 Hepatology Unit, Saint-Eloi Hospital, Montpellier, 5 U871, INSERM, Lyon, 6 Hepatology Unit, Haut Leveque Hospital, Bordeaux, 7 Hepatology Unit, Claude Huriez Hospital, Lille, 8 Medecine Unit, Purpan Hospital, Toulouse, 9 Hepatology Unit, Saint Joseph Hospital, Marseille, 10 Hepatology Unit, Cochin Hospital, 11 Hepatology Unit, Piti´e-Salp´etri`ere Hospital, Paris, 12 Hepatology Unit, Charles Nicolle Hospital, Rouen, 13 Hepatology Unit, Saint-Andr´e Hospital, Bordeaux, 14 Hepatology Unit, Bon Secours Hospital, Metz, 15 Hepatology Unit, Nantes Hospital, Nantes, 16 Hepatology Unit, Purpan Hospital, Toulouse, 17 Virology Unit, Henri Mondor Hospital, Cr´eteil, 18 Hepatology Unit, Paul Brousse Hospital, Villejuif, 19 Hepatology Unit, Brabois Hospital, Nancy, France E-mail: [email protected] Background: Triple therapy was associated with higher SVR rates in phase III trials. However, few patients with cirrhosis were included in these trials. We report the SVR12 rates and safety profile of telaprevir (TVR) and boceprevir (BOC) with PEG-IFN/ribavirin (RBV) in cirrhotic experienced patients treated in the French Early Access Program. Methods: 674 genotype 1 patients with compensated cirrhosis (Child A) were prospectively included and received 12W TVR/PEG-IFN-2a/RBV+36W PEG-IFN/RBV, or 4W PEG-IFN2a/RBV+12W TVR/PEG-IFN-2a/RBV+32W PEG-IFN/RBV, or 4W PEGIFN-2b/RBV+44W BOC/PEG-IFN/RBV. The analysis is restricted to 221 patients reaching W60 of follow-up. Results: Efficacy data are shown in the table. SAEs were observed in 57.0% (TVR) and 40.4% (BOC). Death, infection and hepatic decompensation were reported in 2.8%, 6.5% and 0.9% for TVR and 0.9%, 4.4% and 1.8% for BOC, respectively. Anemia <8 g/dL or blood transfusion were reported in 17.8% (TVR) and 8.7% (BOC). Conclusions: In this large “real life” cohort of cirrhotic patients, SVR12 rate was comparable with the results in the sub group of patients with severe fibrosis or cirrhosis of phase III studies. Data on the entire population included in CUPIC (674 patients) will be presented at the EASL meeting. Table: Virological efficacy Undetectable HCV RNA (ITT) n (%)

BOC n = 114

TVR n = 107

Week 12 Week 24 Week 48 (EOT) SVR12 (Total) SVR12 in relapsers SVR12 in partial responders SVR12 in null responders

52 (46) 58 (51) 44 (39) 44 (39) 24/46 (52) 20/64 (31) 0/2 (0)

79 (74) 65 (61) 45 (42) 49 (46) 32/45 (71) 17/58 (29) 0/3 (0)

Journal of Hepatology 2013 vol. 58 | S25–S44

S27