- Email: [email protected]
Genotypic and Phenotypic Resistance in Clinical Samples Submitted for HCV NS5B Drug Resistance Testing in the Us
POSTER PRESENTATIONS THU-223 NS3 GENETIC VARIABILITY IN HCV GENOTYPE 1A ISOLATES FROM LIVER TISSUE AND SERUM SAMPLES OF TREATMENT-NAÏVE PATIENTS WITH CHRONIC HEPATITIS C D. D’aliberti1, I. Cacciola1, S. Benfatto2, F. Mannino1, R. Filomia1, C. Beninati2, C. Saitta1, G. Raimondo1, T. Pollicino2. 1Clinical and Experimental Medicine; 2Human Pathology, University Hospital of Messina, Messina, Italy E-mail: [email protected] Background and Aims: NS3 protease polymorphisms of HCV genotype (G) 1a potentially conferring resistance to protease inhibitors (PIs) have not been investigated so far in HCV strains from the liver of infected patients where virus replication occurs and HCV quasispecies diversity reaches the highest level of complexity. Aim: To evaluate the frequency of resistance associated variants (RAVs) to NS3 in HCV-G1a isolates from liver tissues and serum samples of patients naïve to any antiviral treatment. Methods: NS3 variability (at amino acid position V36, Q80, R155, A156, D168, V170) of HCV-G1a isolates from paired serum and liver biopsy samples of 11 naïve patients and from serum samples of additional 20 naïve patients was analysed by ultra-deep pyrosequencing (UDPS) (mutant detection sensitivity 1%; >3,000 sequences/patient). Results: UDPS analysis revealed the presence of RAVs in all the 31 studied patients. For single RAVs the detection ratio in liver tissues was higher than in serum samples (median 74% [range 25–100%] vs 50% [range 1–100%], respectively). All but two patients showed RAVs with multiple mutations. D168V/A and I170A were the most frequently detected polymorphisms (found in 24/31 [77%] and 18/ 31 [58%] patients, respectively). The Q80K substitution was found in 12/31 patients (39%), and in 9 of them it was associated with the R155Q/K/T polymorphism. Conclusions: NS3 RAVs are common components of the intrahepatic viral quasispecies. By UDPS they are frequently identified also in circulating viral populations. Considering the very high prevalence of SVR obtained by therapeutic regimens including PIs, NS3 RAVs seem to have a limited impact on response to antiviral treatments. THU-224 GENOTYPIC AND PHENOTYPIC RESISTANCE IN CLINICAL SAMPLES SUBMITTED FOR HCV NS5B DRUG RESISTANCE TESTING IN THE US J.D. Reeves1, J. Volpe1, K. Strommen1, E. Anton1, S. Martens1, D. Yang1, J. Whitcomb1, W. Huang1, C.J. Petropoulos1, C. Walworth1. 1Monogram Biosciences Inc., South San Francisco, United States E-mail: [email protected] Background and Aims: Regimens containing the NS5B inhibitors dasabuvir (DSV) or sofosbuvir (SOF) are approved for the treatment of HCV infection. We characterized the prevalence of DSV and SOF resistance-associated variants (RAVs) in the first 500 clinical samples received for routine NS5B inhibitor resistance testing. Samples with RAVs were further characterized in a phenotypic assay to evaluate replication capacity (RC) and DSV and SOF susceptibility. Methods: The NS5B region was amplified from HCV genotype (GT) 1 samples submitted to Monogram Biosciences (South San Francisco, CA, USA) for resistance analysis. NS5B sequencing was performed using the Illumina MiSeq platform with a 10% variant reporting threshold. DSV and SOF RAVs were identified and an assessment of drug susceptibility (sensitive, resistant (R) or resistance possible (RP)) was derived from a rules-based algorithm. Replicons containing plasma-derived NS5B sequences with RAVs were evaluated for RC and susceptibility to DSV and SOF, relative to the reference GT1b Con1 replicon, using a luciferase-reporter assay. Results: We analyzed the first 500 samples received for routine NS5B resistance testing, of which 83% were GT1a and 17% were GT1b. From genotypic analysis, DSV and/or SOF RAVs were identified in 9.4% of samples overall. For DSV, 7.4% of samples were assessed as R or RP; 7%
for GT1a and 9.3% for GT1b. For SOF, 2.2 % of samples were assessed as R or RP; 2.4% for GT1a and 1.2% for GT1b. From phenotypic analysis, replicons containing NS5B regions from 42 samples with DSV and/or SOF RAVs had RCs ranging from 6-112% and 5–65%, respectively. Susceptibility to DSV and SOF varied over a >2000- and 23-fold range, respectively. GT1a viruses exhibited larger reductions in DSV susceptibility compared to GT1b viruses, with the greatest reductions in susceptibility (IC50FC > 975) seen for samples with C316Y variants. For SOF, the largest reductions in susceptibility (IC50FC of up to 12) were seen among GT1a viruses with S282T and L159F variants. Conclusions: From a survey of samples submitted for NS5B resistance testing, DSV RAVs were more prevalent compared to SOF RAVs. Overall, RAV prevalence was similar among GT1a and GT1b viruses. Susceptibility to DSV and SOF varied over an approximate 3 and 1 log range, respectively, with the largest reductions in inhibitor susceptibility seen among viruses with GT1a RAVs. THU-225 C-EDGE CO-STAR: FAVORABLE IMPACT OF ELBASVIR AND GRAZOPREVIR ON HEALTH-RELATED QUALITY OF LIFE IN TREATMENT-NAÏVE HCV-INFECTED PERSONS WHO INJECT DRUGS RECEIVING OPIOID AGONIST THERAPY J.M. Arduino1, O. Shibolet2, A.H. Litwin3, J. Grebeley4, F. Altice5, C. Nwankwo1, T.C. Mast1, Z. Jiang1, I.N. Gendrano1, H.L. Platt1, G.J. Dore4. 1Merck & Co., Inc., Kenilworth, NJ, United States; 2Tel-Aviv Medical Center and Tel-Aviv University, Tel-Aviv, Israel; 3Montefiore Medical Center and Albert Einstein College of Medicine, Bronx, NY, United States; 4The Kirby Institute, UNSW, Sydney, Australia; 5Yale School of Medicine, New Haven, CT, United States E-mail: [email protected] Background and Aims: A Phase 3, double-blind, placebo-control, randomized trial of an oral fixed dose combination of elbasvir (EBR, an HCV NS5A inhibitor) 50 mg / grazoprevir (GZR, an HCV NS3/4A protease inhibitor) 100 mg once daily for 12 weeks was conducted among treatment-naïve HCV GT1-,4-,or 6-infected patients who inject drugs receiving opioid agonist therapy (OAT). EBR/GZR was demonstrated to be highly effective (SVR12: 95.5% (95% CI: 91.5%, 97.9%)) and generally well-tolerated, with a similar safety profile to placebo. The aim of this study was to evaluate whether HCV treatment with EBR/GZR altered the health-related quality of life (HRQOL) profile in patients on OAT. Methods: HRQOL was assessed using the SF-36V2® Acute Health Survey. Patients completed the SF-36V2® at baseline, treatment week 4 (TW4), TW12, follow-up week 4 (FW4), and FW12 (EBR/GZR arm only). 301 patients were randomized and received ≥1 dose of study drug (EBR/GZR: n = 201, Placebo: n = 100). The aim of this study was to estimate the mean changes in health domains’, mental component summary (MCS) and physical component summary (PCS) scores from baseline to TW12 by treatment group.
Journal of Hepatology 2016 vol. 64 | S213–S424
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for GT1a and 9.3% for GT1b. For SOF, 2.2 % of samples were assessed as R or RP; 2.4% for GT1a and 1.2% for GT1b. From phenotypic analysis, replicons containing NS5B regions from 42 samples with DSV and/or SOF RAVs had RCs ranging from 6-112% and 5–65%, respectively. Susceptibility to DSV and SOF varied over a >2000- and 23-fold range, respectively. GT1a viruses exhibited larger reductions in DSV susceptibility compared to GT1b viruses, with the greatest reductions in susceptibility (IC50FC > 975) seen for samples with C316Y variants. For SOF, the largest reductions in susceptibility (IC50FC of up to 12) were seen among GT1a viruses with S282T and L159F variants. Conclusions: From a survey of samples submitted for NS5B resistance testing, DSV RAVs were more prevalent compared to SOF RAVs. Overall, RAV prevalence was similar among GT1a and GT1b viruses. Susceptibility to DSV and SOF varied over an approximate 3 and 1 log range, respectively, with the largest reductions in inhibitor susceptibility seen among viruses with GT1a RAVs. THU-225 C-EDGE CO-STAR: FAVORABLE IMPACT OF ELBASVIR AND GRAZOPREVIR ON HEALTH-RELATED QUALITY OF LIFE IN TREATMENT-NAÏVE HCV-INFECTED PERSONS WHO INJECT DRUGS RECEIVING OPIOID AGONIST THERAPY J.M. Arduino1, O. Shibolet2, A.H. Litwin3, J. Grebeley4, F. Altice5, C. Nwankwo1, T.C. Mast1, Z. Jiang1, I.N. Gendrano1, H.L. Platt1, G.J. Dore4. 1Merck & Co., Inc., Kenilworth, NJ, United States; 2Tel-Aviv Medical Center and Tel-Aviv University, Tel-Aviv, Israel; 3Montefiore Medical Center and Albert Einstein College of Medicine, Bronx, NY, United States; 4The Kirby Institute, UNSW, Sydney, Australia; 5Yale School of Medicine, New Haven, CT, United States E-mail: [email protected] Background and Aims: A Phase 3, double-blind, placebo-control, randomized trial of an oral fixed dose combination of elbasvir (EBR, an HCV NS5A inhibitor) 50 mg / grazoprevir (GZR, an HCV NS3/4A protease inhibitor) 100 mg once daily for 12 weeks was conducted among treatment-naïve HCV GT1-,4-,or 6-infected patients who inject drugs receiving opioid agonist therapy (OAT). EBR/GZR was demonstrated to be highly effective (SVR12: 95.5% (95% CI: 91.5%, 97.9%)) and generally well-tolerated, with a similar safety profile to placebo. The aim of this study was to evaluate whether HCV treatment with EBR/GZR altered the health-related quality of life (HRQOL) profile in patients on OAT. Methods: HRQOL was assessed using the SF-36V2® Acute Health Survey. Patients completed the SF-36V2® at baseline, treatment week 4 (TW4), TW12, follow-up week 4 (FW4), and FW12 (EBR/GZR arm only). 301 patients were randomized and received ≥1 dose of study drug (EBR/GZR: n = 201, Placebo: n = 100). The aim of this study was to estimate the mean changes in health domains’, mental component summary (MCS) and physical component summary (PCS) scores from baseline to TW12 by treatment group.
Journal of Hepatology 2016 vol. 64 | S213–S424
S403