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      Prevalence of Single and Multiple Natural NS3, NS5A and NS5B Resistance-Associated Substitutions in Hepatitis C Virus Genotypes 1–4 in Italy

      research-article
      1 , 1 , 1 , 2 , 3 , 4 , 1 , 5 , 6 , 2 , 7 , 8 , 9 , 2 , 4 , 3 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 7 , 6 , 5 , 3 , 4 , 2 , 19 , 1 , , 1 , HCV Virology Italian Resistance Network (VIRONET-C)
      Scientific Reports
      Nature Publishing Group UK

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          Abstract

          Natural resistance-associated substitutions (RASs) are reported with highly variable prevalence across different HCV genotypes (GTs). Frequency of natural RASs in a large Italian real-life cohort of patients infected with the 4 main HCV-GTs was investigated. NS3, NS5A and NS5B sequences were analysed in 1445 HCV-infected DAA-naïve patients. Sanger-sequencing was performed by home-made protocols on 464 GT1a, 585 GT1b, 92 GT2c, 199 GT3a, 16 GT4a and 99 GT4d samples. Overall, 20.7% (301/1455) of patients showed natural RASs, and the prevalence of multiclass-resistance was 7.3% (29/372 patients analysed). NS3-RASs were particularly common in GT1a and GT1b (45.2-10.8%, respectively), mainly due to 80K presence in GT1a (17%). Almost all GTs showed high prevalence of NS5A-RASs (range: 10.2–45.4%), and especially of 93H (5.1%). NS5A-RASs with fold-change >100x were detected in 6.8% GT1a (30H/R-31M-93C/H), 10.3% GT1b (31V-93H), 28.4% GT2c (28C-31M-93H), 8.5% GT3a (30K-93H), 45.5% GT4a (28M-30R-93H) and 3.8% GT4d (28V-30S-93H). Sofosbuvir RAS 282T was never detected, while the 159F and 316N RASs were found in GT1b (13.4–19.1%, respectively). Natural RASs are common in Italian patients infected with HCV-GTs 1–4. High prevalence of clinically-relevant RASs (such as Y93H) supports the appropriateness of HCV resistance-test to properly guide DAA-based therapy.

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          Most cited references32

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          Glecaprevir and pibrentasvir yield high response rates in patients with HCV genotype 1-6 without cirrhosis.

          Hepatitis C virus (HCV) therapy that is highly efficacious, pangenotypic, with a high barrier to resistance and short treatment duration is desirable. The efficacy and safety of 8- and 12-week treatments with glecaprevir (ABT-493; NS3/4A protease inhibitor) and pibrentasvir (ABT-530; NS5A inhibitor) were evaluated in non-cirrhotic patients with chronic HCV genotype 1-6 infection.
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            Efficacy and safety of 12 weeks versus 18 weeks of treatment with grazoprevir (MK-5172) and elbasvir (MK-8742) with or without ribavirin for hepatitis C virus genotype 1 infection in previously untreated patients with cirrhosis and patients with previous null response with or without cirrhosis (C-WORTHY): a randomised, open-label phase 2 trial.

            There is a high medical need for an interferon-free, all-oral, short-duration therapy for hepatitis C virus (HCV) that is highly effective across diverse patient populations, including patients with cirrhosis or previous null response to pegylated interferon (peginterferon) plus ribavirin (PR-null responders). We aimed to assess the efficacy, safety, and effective treatment duration of grazoprevir (an HCV NS3/4A protease inhibitor) combined with elbasvir (an HCV NS5A inhibitor) with or without ribavirin in patients with HCV genotype 1 infection with baseline characteristics of poor response. The C-WORTHY trial is a randomised, open-label phase 2 trial of grazoprevir plus elbasvir with or without ribavirin; here we report findings for two cohorts of previously untreated patients with cirrhosis (cohort 1) and those with previous PR-null response with or without cirrhosis (cohort 2) enrolled in part B of the study. Eligible patients were adults aged 18 years or older with chronic HCV genotype 1 infection and HCV RNA concentrations of 10 000 IU/mL or higher in peripheral blood. We randomly assigned patients to receive grazoprevir (100 mg daily) and elbasvir (50 mg daily) with or without ribavirin for 12 or 18 weeks. Randomisation was done centrally with an interactive voice response system; patients and study investigators were masked to treatment duration up to week 12 but not to treatment allocation. The primary endpoint was the proportion of patients achieving HCV RNA less than 25 IU/mL at 12 weeks after end of treatment (SVR12), assessed by COBAS TaqMan version 2.0. This study is registered with ClinicalTrials.gov, number NCT01717326. We describe findings for 253 patients enrolled in cohort 1 (n=123) or cohort 2 (n=130). In cohort 1, we randomly assigned 60 patients to the 12-week regimen (31 with ribavirin and 29 with no ribavirin) and 63 to the 18-week regimen (32 with ribavirin and 31 with no ribavirin); in cohort 2, we randomly assigned 65 patients to the 12-week regimen (32 with ribavirin and 33 with no ribavirin) and 65 to the 18-week regimen (33 with ribavirin and 32 with no ribavirin. High SVR12 rates were achieved irrespective of the use of ribavirin or extension of the treatment duration from 12 to 18 weeks; SVR12 rates ranged from 90% (95% CI 74-98; 28/31; cohort 1, 12 weeks, ribavirin-containing) to 100% (95% CI 89-100; 33/33; cohort 2, 18 weeks, ribavirin-containing). Among patients treated for 12 weeks with grazoprevir plus elbasvir without ribavirin, 97% (95% CI 82-100, 28/29) of patients in cohort 1 and 91% (76-98, 30/33) of patients in cohort 2 achieved SVR12. Adverse events reported in more than 10% of patients were fatigue (66 patients, 26% [95% CI 21-32]), headache (58 patients, 23% [95% CI 18-29]), and asthenia (35 patients, 14% [95% CI 10-19]). Treatment with grazoprevir plus elbasvir, both with and without ribavirin and for both 12 and 18 weeks' treatment duration, showed high rates of efficacy in previously untreated patients with cirrhosis and previous PR-null responders with and without cirrhosis. These results support the phase 3 development of grazoprevir plus elbasvir. Merck & Co, Inc. Copyright © 2015 Elsevier Ltd. All rights reserved.
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              Efficacy of 8 Weeks of Sofosbuvir, Velpatasvir, and Voxilaprevir in Patients With Chronic HCV Infection: 2 Phase 3 Randomized Trials.

              Patients with chronic hepatitis C virus (HCV) infection have high rates of sustained virologic response (SVR) after 12 weeks of treatment with the nucleotide polymerase inhibitor sofosbuvir combined with the NS5A inhibitor velpatasvir. We assessed the efficacy of 8 weeks of treatment with sofosbuvir and velpatasvir plus the pangenotypic NS3/4A protease inhibitor voxilaprevir (sofosbuvir-velpatasvir-voxilaprevir).

                Author and article information

                Contributors
                valeriacento@gmail.com
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                12 June 2018
                12 June 2018
                2018
                : 8
                : 8988
                Affiliations
                [1 ]ISNI 0000 0001 2300 0941, GRID grid.6530.0, Department Experimental Medicine and Surgery, , University of Rome “Tor Vergata”, ; 00133 Rome, Italy
                [2 ]Hepatology Unit, University Hospital of Rome “Tor Vergata”, 00133 Rome, Italy
                [3 ]Infectious Diseases Unit, University Hospital of Rome “Tor Vergata”, 00133 Rome, Italy
                [4 ]GRID grid.461844.b, Infectious Diseases Unit, , Pescara General Hospital, ; 65124 Pescara, Italy
                [5 ]ISNI 0000 0004 1757 123X, GRID grid.415230.1, Infectious Diseases Unit, , Sant’Andrea Hospital – “Sapienza” University, ; 00189 Rome, Italy
                [6 ]GRID grid.417007.5, Tropical Diseases, , Umberto I Hospital –“Sapienza” University, ; 00161 Rome, Italy
                [7 ]1st Division of Infectious Diseases, ASST Fatebenefratelli Sacco, 20157 Milan, Italy
                [8 ]ISNI 0000 0001 2097 9138, GRID grid.11450.31, Clinical and Experimental Medicine, , University of Sassari, ; 07100 Sassari, Italy
                [9 ]ISNI 0000 0004 1756 7871, GRID grid.410345.7, Infectious Diseases Unit, , IRCCS AOU San Martino - IST, ; 16132 Genoa, Italy
                [10 ]ISNI 0000 0004 1805 3485, GRID grid.416308.8, Hepatology Unit, , San Camillo Forlanini Hospital, ; 00151 Rome, Italy
                [11 ]ISNI 0000 0004 1760 3027, GRID grid.419425.f, Molecular Virology, , Fondazione IRCCS Policlinico San Matteo, ; 27100 Pavia, Italy
                [12 ]Gastroenterology, “P. Giaccone” University Hospital, 90127 Palermo, Italy
                [13 ]ISNI 0000 0001 0790 385X, GRID grid.4691.a, Gastroenterology, , “Federico II” University, ; 80131 Naples, Italy
                [14 ]GRID grid.461844.b, Hepatology Unit, , Pescara General Hospital, ; 65124 Pescara, Italy
                [15 ]ISNI 0000 0001 0941 3192, GRID grid.8142.f, Gastroenterology, , Catholic University of Rome, ; 00168 Rome, Italy
                [16 ]Infectious Diseases Unit, “L. Vanvitelli” University of Campania, 80138 Naples, Italy
                [17 ]Infectious Diseases Unit, S. Salvatore Hospital, 67100 L’Aquila, Italy
                [18 ]GRID grid.416200.1, Infectious Diseases Unit, , Niguarda Ca’ Granda Hospital, ; 20162 Milan, Italy
                [19 ]ISNI 0000 0004 1757 2822, GRID grid.4708.b, Haematology and Oncohematology, , University of Milan, ; 20122 Milan, Italy
                [20 ]ISNI 0000 0004 1757 1758, GRID grid.6292.f, Centro di Ricerca per lo Studio delle Epatiti, , University of Bologna, ; 40138 Bologna, Italy
                [21 ]ISNI 0000 0004 1760 3027, GRID grid.419425.f, Infectious and Tropical Diseases, , Fondazione IRCCS Policlinico San Matteo, ; 27100 Pavia, Italy
                [22 ]ISNI 0000 0004 1756 8807, GRID grid.417728.f, Humanitas University and IRCCS Istituto Clinico Humanitas, ; 20089 Rozzano, MI Italy
                [23 ]ISNI 0000 0001 2336 6580, GRID grid.7605.4, Infectious Diseases Unit, Amedeo di Savoia Hospital, , University of Turin, ; 10149 Turin, Italy
                [24 ]Clinic of Infectious Diseases, AOUM, 41125 Modena, Italy
                [25 ]ISNI 0000 0004 1756 7871, GRID grid.410345.7, Hygiene Unit, , IRCCS AOU San Martino - IST, ; 16132 Genoa, Italy
                [26 ]ISNI 0000 0001 2174 1754, GRID grid.7563.7, Center for Digestive Health, , University of Milano-Bicocca, ; 20900 Monza, Italy
                [27 ]ISNI 0000 0001 2336 6580, GRID grid.7605.4, Gastroenterology and Hepatology, , University of Turin, ; 10126 Turin, Italy
                [28 ]Hepatology Unit, AOUP, Ospedale Nuovo Santa Chiara, Cisanello, 56124 Pisa Italy
                [29 ]Gastroenterology, “De Bellis” Hospital, IRCCS, 70013 Castellana Grotte, BA Italy
                [30 ]GRID grid.417007.5, Infectious Diseases Unit, , “Sapienza” University of Rome, ; 00161 Rome, Italy
                [31 ]ISNI 0000 0004 1759 0844, GRID grid.411477.0, Infectious Diseases Unit, , Siena University Hospital, ; 53100 Siena, Italy
                [32 ]ISNI 0000 0004 1763 1028, GRID grid.413671.6, Microbiology and Virology Laboratory, , Amedeo di Savoia Hospital, ; 10126 Turin, Italy
                [33 ]ISNI 0000 0004 1757 2822, GRID grid.4708.b, Unit San Paolo School of Medicine Department of Health Sciences, , University of Milan, ; 20142 Milan, Italy
                [34 ]ISNI 0000 0004 1760 3027, GRID grid.419425.f, Infectious Diseases Unit, , Fondazione IRCCS Policlinico San Matteo, ; 27100 Pavia, Italy
                [35 ]GRID grid.7841.a, Infectious Diseases Unit, , Sapienza University, ; 04100 Latina, Italy
                [36 ]ISNI 0000 0004 1756 7871, GRID grid.410345.7, Gastroenterology Unit, , IRCCS AOU San Martino - IST, ; 16132 Genoa, Italy
                [37 ]GRID grid.417176.2, Infectious Diseases Unit, , Ospedale di circolo di Busto Arsizio, ; 21052 Busto Arsizio, VA Italy
                [38 ]Clinical Microbiology, Virology and Bioemergencies, ASST Fatebenefratelli Sacco, 20157 Milan, Italy
                [39 ]Infectious Disease Unit, Avezzano General Hospital, 67051 Avezzano, AQ Italy
                [40 ]ISNI 0000 0001 0120 3326, GRID grid.7644.1, Gastroenterology Unit, , University of Bari, ; 70121 Bari Bari, Italy
                [41 ]ISNI 0000000121697570, GRID grid.7548.e, Department of Biomedical, Metabolic and Neural Sciences, , University of Modena and Reggio Emilia, ; 41125 Modena, Italy
                [42 ]Third Infectious Disease Unit, A.O. Cotugno, 80131 Naples, Italy
                [43 ]Infectious Disease Unit, Hospital “G. Mazzini”, 64100 Teramo, Italy
                [44 ]ISNI 0000 0001 2113 062X, GRID grid.5390.f, Department of Experimental and Clinical Medicine, , University of Udine, ; 33100 Udine, Italy
                [45 ]ISNI 0000 0004 1757 4641, GRID grid.9024.f, Department of Medical Biotechnology, , University of Siena, ; 53100 Siena, Italy
                Author information
                http://orcid.org/0000-0003-0663-2503
                Article
                26862
                10.1038/s41598-018-26862-y
                5997636
                29895871
                ebce5a11-fb6c-47aa-a730-ce265b8aad62
                © The Author(s) 2018

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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                : 3 January 2018
                : 16 May 2018
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