- Email: [email protected]
Molecular characterization of Hepatitis C virus resistance-associated substitutions after interferon-free treatment failure by massive parallel sequencing
POSTER PRESENTATIONS Conclusions: Based on recommendations, retreatment of patients after a first DAAs regimen failure is effectiveness (95% SVR). However, despite the strong efficiency of this strategy guidance therapy, some limits can be observed in some few “difficult-to-cure” patients. Full genome resistance analysis or future triple DAA therapy is mandated in these patients in order to choose the optimal combination therapy. SAT-256 Molecular characterization of Hepatitis C virus resistanceassociated substitutions after interferon-free treatment failure by massive parallel sequencing Q. Chen1, C. Perales1, J. Crespo2, M. Buti3, J.L. Calleja4, J. Gregori1, F. Rodríguez-Frías1, L. Nieto3, J. Niubó5, M. Rodríguez6, I. Conde7, M.L.G. Buey8, I. Fernández9, J. Torras10, R. Morillas11, M. Diago12, F. Sáez-Royuela13, M. Forné14, J. Tunes15, J.G. Samaniego16, J.A. Carrión17, J.I. Arenas18, X. Forns19, Z. Mariño19, R. Andrade20, S. Montoliu21, M.Á. Simón22, G.S. Antolín23, J.J. Sánchez-Ruano24, J.M. Pascasio25, J.I. Esteban3, J. Quer1. 1Vall d’Hebron Institut de Recerca – Hospital Universitari Vall d’Hebron, Barcelona; 2Hospital Universitario Marqués de Valdecilla – Instituto de Investigación Valdecilla, Santander; 3 Hospital Universitari Vall d’Hebron, Barcelona; 4Hospital Universitario Puerta de Hierro, Madrid; 5Hospital Universitari de Bellvitge, Hospitalet de Llobregat; 6Hospital Central Universitario Asturias, Barcelona; 7 Hospital Universitario La Fe, Valencia; 8Hospital Universitario La Princesa; 9Hospital Universitario 12 de Octubre, Madrid; 10Hospital de la Santa Cruz y San Pablo, Barcelona; 11Hospital Universitario Germans Trias i Pujol, Badalona; 12Hospital General de Valencia, Valencia; 13 Hospital Universitario Burgos, Burgos; 14Hospital Mútua de Terrassa, Terrassa; 15Hospital Universitario de Pontevedra, Pontevedra; 16Hospital Universitario La Paz, Madrid; 17Hospital del Mar, Barcelona; 18Hospital Universitario de Donostia, San Sebastián; 19Hospital Clínic de Barcelona, Barcelona; 20Hospital Virgen de la Victoria, Málaga; 21Hospital Universitari Joan XXIII de Tarragona, Tarragona; 22Hospital Universitario Miguel Servet, Zaragoza; 23Hospital Río Hortega, Valladolid; 24Hospital Universitario de Toledo, Toledo; 25Hospital Universitario Virgen del Rocío, Sevilla, Spain E-mail: [email protected] Background and Aims: At present, more than 60.000 chronically Hepatitis C infected patients have started Direct-Acting Antiviral (DAA) Interferon-free treatments in Spain. It has been estimated that around 2% will fail and viral resistant-associated substitutions (RASs) are usually selected, thus conditioning future therapeutic options due to the cross-resistance with inhibitors of the same family. In this scenario, the best accurate retreatment strategy depends on the type, frequency and how RASs are combined in the same genome. The aim of the study is to detect RASs among treatment failure patients, the frequency at which they are found and their combination in the same genome using massive parallel sequencing. Methods: More than 150 treatment failure patients were recruited from 29 Spanish hospitals included in the National HCV Strategic Plan with collaboration of CIBER of Liver and Digestive Diseases (Ciberehd). All serum samples were genotyped and RASs were analyzed by Next Generation Sequencing (NGS). Results: The vast majority of patients got a relapse of viral infection once several weeks after stopping DAA treatment. 90% of the patients show RASs in at least one analyzed regions (NS3, NS5A and/or NS5B). The subtype distribution among all treatment failure samples was 53% G1b, 16% G1a, 17% G3a, 9% G4d, 1% G2j, 1% G2c, 1% G4a and 2% mixed infections. The most prevalent RAS were detected in positions 80, 122, 155 and 168 in NS3 region, positions 28, 30, 58 and 93 in NS5A region, positions 159 and 316 in NS5B region. Moreover, resistance substitution S282T, which has been directly associated with resistance to Sofosbuvir (SOF), despite causing a dramatic viral fitness decay in cell culture experiments, has been detected in 3% of SOF treated patients. The combination Q80R + D168E and L31M + Y93H, have been shown in 6.5% and 14.5% of G1b patients,
respectively. Among all analyzed samples, 18% of patients did not show any RAS despite relapse of viral load in plasma. Conclusions: The extensive characterization of the spectrum of substitutions by massive parallel sequencing is an essential tool to identify resistance-associated substitutions, combination of variants in the same sequence and their frequency during direct-acting antiviral based treatment failures. This information will help design the most accurate re-treatment option for these patients. SAT-257 Improved virological outcomes and excellent safety profile in genotype 3 HCV-infected cirrhotic patients after an extended 24weeks course of daclatasvir, sofosbuvir + ribavirin: insights from a real-life multicenter study R. Lionetti1, I. Lenci2, M. Siciliano3, M. Pompili3, U.V. Comandini1, M. Milana2, C. Taibi1, M. Montalbano1, A.R. Garbuglia4, P. Begini5, B. Imperatrice6, M. Angelico2, G. D’Offizi1. 1Liver and Infectious Diseases, Lazzaro Spallanzani; 2Liver Unit, Policlinico Tor Vergata; 3 Gastroenterology and Liver Unit, Policlinico Gemelli, Catholic university; 4Virology, Lazzaro Spallanzani; 5Liver Unit, Sant’Andrea University Hospital; 6Liver unit, Sabt’Andrea University Hospital, Rome, Italy E-mail: [email protected] Background and Aims: Patients with HCV genotype 3 (G3) infection, expecially if cirrhotics, show unsatisfactory virological outcomes after conventional 12 weeks treatment with direct antiviral agents (DAA). The ALLY-3 Plus study showed that treatment extension to 16 weeks was not associated with improved sustained virological response (SVR12), which remained <90% in cirrhotic patients. The National Italian Drug Agency rules allow prescription of 24 weeks of DAA therapy in most difficult-to-treat patients, including cirrhotics with G3 infection. To evaluate the virological efficacy and safety of an extended 24-week treatment with a combination of Daclatasvir (DCV), Sofosbuvir (SOF) ± Ribavirin (RBV) in G3 cirrhotic patients. Methods: Seventy-four consecutive patients (15F, 59M) received the above DAA combination for 24 weeks. The use of RBV was based on patients’ clinical features and/or expected tolerability. RBV was administered in 54 (72.9%) pts, at a median dose of 922 ± 200 mg/ day. Forty-seven patients were treatment-naïve and 27 had failed after interferon-alfa and RBV therapy (12 relapsers and 15 nonresponders). BMI was <26 in 37, 26–30 in 30 and >30 in 7 pts. At baseline, disease severity, assessed by Child-Pugh, showed stage A in 65, stage B in 7 and stage C in 2, with a mean MELD score of 9.2 ± 2.9. Baseline ALT values were 113 ± 96 IU and HCVRNA levels were <800.000 IU in 47 and >800.000 IU in 27 pts. Results: Sixty-eight patients reached a 12-week follow-up after endof treatment, attaining a SVR12 rate of 97% (n = 66). Interestingly, SVR12 was 100% among RBV treated patients vs. 90% (18/20) among those treated without RBV. Of the remaining 6 treated patients all but one achieved SVR4. No patients discontinued DAAs. The only relevant side effect was anemia, reported in the RBV group only, and easily managed by dose reductions. No changes in renal and liver function were recorded baseline vs. follow up. The two patients who failed (one previous non-responder and one treatment-naïve) showed the development of resistance associated variants (Y93H in NS5A in both cases and S282T in one only). Conclusions: An extended 24-week treatment schedule with the triple combination of DCV, SOF and RBV allows to achieve a 100% SVR12 rate in the difficult-to-treat group of HCV G3-infected cirrhotics, with very limited side effects. The role of RBV seems to be crucial in this particular setting and should be always administered if clinical conditions allow it, although larger confirmatory studies are needed.
Journal of Hepatology 2017 vol. 66 | S543–S750
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respectively. Among all analyzed samples, 18% of patients did not show any RAS despite relapse of viral load in plasma. Conclusions: The extensive characterization of the spectrum of substitutions by massive parallel sequencing is an essential tool to identify resistance-associated substitutions, combination of variants in the same sequence and their frequency during direct-acting antiviral based treatment failures. This information will help design the most accurate re-treatment option for these patients. SAT-257 Improved virological outcomes and excellent safety profile in genotype 3 HCV-infected cirrhotic patients after an extended 24weeks course of daclatasvir, sofosbuvir + ribavirin: insights from a real-life multicenter study R. Lionetti1, I. Lenci2, M. Siciliano3, M. Pompili3, U.V. Comandini1, M. Milana2, C. Taibi1, M. Montalbano1, A.R. Garbuglia4, P. Begini5, B. Imperatrice6, M. Angelico2, G. D’Offizi1. 1Liver and Infectious Diseases, Lazzaro Spallanzani; 2Liver Unit, Policlinico Tor Vergata; 3 Gastroenterology and Liver Unit, Policlinico Gemelli, Catholic university; 4Virology, Lazzaro Spallanzani; 5Liver Unit, Sant’Andrea University Hospital; 6Liver unit, Sabt’Andrea University Hospital, Rome, Italy E-mail: [email protected] Background and Aims: Patients with HCV genotype 3 (G3) infection, expecially if cirrhotics, show unsatisfactory virological outcomes after conventional 12 weeks treatment with direct antiviral agents (DAA). The ALLY-3 Plus study showed that treatment extension to 16 weeks was not associated with improved sustained virological response (SVR12), which remained <90% in cirrhotic patients. The National Italian Drug Agency rules allow prescription of 24 weeks of DAA therapy in most difficult-to-treat patients, including cirrhotics with G3 infection. To evaluate the virological efficacy and safety of an extended 24-week treatment with a combination of Daclatasvir (DCV), Sofosbuvir (SOF) ± Ribavirin (RBV) in G3 cirrhotic patients. Methods: Seventy-four consecutive patients (15F, 59M) received the above DAA combination for 24 weeks. The use of RBV was based on patients’ clinical features and/or expected tolerability. RBV was administered in 54 (72.9%) pts, at a median dose of 922 ± 200 mg/ day. Forty-seven patients were treatment-naïve and 27 had failed after interferon-alfa and RBV therapy (12 relapsers and 15 nonresponders). BMI was <26 in 37, 26–30 in 30 and >30 in 7 pts. At baseline, disease severity, assessed by Child-Pugh, showed stage A in 65, stage B in 7 and stage C in 2, with a mean MELD score of 9.2 ± 2.9. Baseline ALT values were 113 ± 96 IU and HCVRNA levels were <800.000 IU in 47 and >800.000 IU in 27 pts. Results: Sixty-eight patients reached a 12-week follow-up after endof treatment, attaining a SVR12 rate of 97% (n = 66). Interestingly, SVR12 was 100% among RBV treated patients vs. 90% (18/20) among those treated without RBV. Of the remaining 6 treated patients all but one achieved SVR4. No patients discontinued DAAs. The only relevant side effect was anemia, reported in the RBV group only, and easily managed by dose reductions. No changes in renal and liver function were recorded baseline vs. follow up. The two patients who failed (one previous non-responder and one treatment-naïve) showed the development of resistance associated variants (Y93H in NS5A in both cases and S282T in one only). Conclusions: An extended 24-week treatment schedule with the triple combination of DCV, SOF and RBV allows to achieve a 100% SVR12 rate in the difficult-to-treat group of HCV G3-infected cirrhotics, with very limited side effects. The role of RBV seems to be crucial in this particular setting and should be always administered if clinical conditions allow it, although larger confirmatory studies are needed.
Journal of Hepatology 2017 vol. 66 | S543–S750
S731