Detection of anti-protease inhibitors resistance mutations in HCV strains infecting treatment-naïve chronic patients from Romania

Open access

Abstract

Background: Severe complications of chronic hepatitis C – i.e. cirrhosis and hepatocellular carcinoma – are important causes of morbidity and mortality worldwide. Despite the overwhelming rates of sustained virologic response achieved after therapy with different combinations of direct-acting antiviral drugs (DAAs), treatment failure is still recorded, and is due to the mutations harboured by hepatitis C virus (HCV) resistance associated variants (RAVs) selected during therapy. Baseline RAVs testing was found significant for guiding treatment in the cases of treatment failure and, sometimes, in naïve patients.

Methods: Romanian chronic hepatitis C patients unexposed to DAAs and infected with subtype 1b HCV were studied. Serum samples were used for Sanger population sequencing of a fragment containing NS3 viral protease, known to harbour resistance mutation against protease inhibitors (PIs).

Results: Catalytic triad and zinc-binding site in the studied sequences were conserved. Low-intermediate resistance mutations to first generation PIs were detected either alone or in conjunction with resistance substitutions associated with second generation PIs. Cross-resistance and reduced susceptibility to certain DAAs were observed.

Discussion: This study focused on HCV patients infected with subtype 1b strains, the most prevalent in Romania. The rate of RAVs found in this work is consistent with the results reported by similar studies from other countries. Noticeably, numerous polymorphisms of unknown significance to DAAs resistance, but reflecting the high genetic variability of HCV, were found in the studied sequences. Testing for RAVs can be a useful method for guiding treatment in a cost-efficient manner in developing countries where access to DAAs is limited.

1. Chen SL, Morgan TR. The natural history of hepatitis C virus (HCV) infection. Int J Med Sci. 2006;3(2):47-52. DOI: 10.7150/ijms.3.47

2. Fields BN, Knipe DM, Howley PM. Fields virology. 5th ed. Philadelphia: Wolters Kluwer Health/Lippincott Williams & Wilkins. 2007.

3. Smith DB, Bukh J, Kuiken C, Muerhoff AS, Rice CM, Stapleton JT, et al. Expanded classification of hepatitis C virus into 7 genotypes and 67 subtypes: updated criteria and genotype assignment web resource. Hepatology. 2014;59(1):318-27. DOI: 10.1002/hep.26744

4. Simmonds P, Becher P, Bukh J, Gould EA, Meyers G, Monath T, et al. ICTV Virus Taxonomy Profile: Flaviviridae. J Gen Virol. 2017;98(1):2-3. DOI: 10.1099/jgv.0.000672

5. Smith DB, Becher P, Bukh J, Gould EA, Meyers G, Monath T, et al. Proposed update to the taxonomy of the genera Hepacivirus and Pegivirus within the Flaviviridae family. J Gen Virol. 2016;97(11):2894-907. DOI: 10.1099/jgv.0.000612

6. Sherman KE, Flamm SL, Afdhal NH, Nelson DR, Sulkowski MS, Everson GT, et al. Response-guided telaprevir combination treatment for hepatitis C virus infection. N Engl J Med. 2011;365(11):1014-24. DOI: 10.1056/NEJMoa1014463

7. Poordad F, McCone J, Jr., Bacon BR, Bruno S, Manns MP, Sulkowski MS, et al. Boceprevir for untreated chronic HCV genotype 1 infection. N Engl J Med. 2011;364(13):1195-206. DOI: 10.1056/NEJMoa1010494

8. European Association for Study of Liver. EASL Recommendations on Treatment of Hepatitis C 2015. J Hepatol. 2015;63(1):199-236. DOI: 10.1016/j.jhep.2015.03.025

9. Macartney MJ, Irish D, Bridge SH, Garcia-Diaz A, Booth CL, McCormick AL, et al. Telaprevir or boceprevir based therapy for chronic hepatitis C infection: development of resistance-associated variants in treatment failure. Antiviral Res. 2014;105:112-7. DOI: 10.1016/j.antiviral.2014.02.019

10. Sarrazin C. The importance of resistance to direct antiviral drugs in HCV infection in clinical practice. J Hepatol. 2016;64(2):486-504. DOI: 10.1016/j.jhep.2015.09.011

11. Hezode C, Asselah T, Reddy KR, Hassanein T, Berenguer M, Fleischer-Stepniewska K, et al. Ombitasvir plus paritaprevir plus ritonavir with or without ribavirin in treatment-naive and treatment-experienced patients with genotype 4 chronic hepatitis C virus infection (PEARL-I): a randomised, open-label trial. Lancet. 2015. DOI: 10.1016/S0140-6736(15)60159-3

12. Gheorghe L, Iacob S, Simionov I, Caruntu F, Motoc A, Arama V, et al. A real life boceprevir use in treatment-experienced HCV genotype 1 patients with advanced fibrosis. J Gastrointestin Liver Dis. 2014;23(1):45-50.

13. Oprisan G, Dinu S, Micu L, Micu G, Ecobici M, Spandole S, et al. Study of genetic and viral markers associated with nonresponse to triple therapy for patients with genotype 1 chronic hepatitis C. International Conference “Education and creativity for a knowledge-based society”, November 17-19, 2016. Bucharest.

14. Leblebicioglu H, Arends JE, Ozaras R, Corti G, Santos L, Boesecke C, et al. Availability of hepatitis C diagnostics and therapeutics in European and Eurasia countries. Antiviral Res. 2018;150:9-14. DOI: 10.1016/j.antiviral.2017.12.001

15. Nițescu M, Vâjâitu C, Săndulescu O, Streinu-Cercel A, Pițigoi D, Preoțescu L, et al. Non-invasive quantification of liver fibrosis regression following successful treatment of chronic hepatitis C with direct acting antivirals. Rev Romana Med Lab. 2017;25(4):355-63. DOI: 10.1515/rrlm-2017-0030

16. Sultana C, Oprisan G, Szmal C, Vagu C, Temereanca A, Dinu S, et al. Molecular epidemiology of hepatitis C virus strains from Romania. J Gastrointestin Liver Dis. 2011;20(3):261-6.

17. Sultana C, Oprisan G, Teleman MD, Dinu S, Oprea C, Voiculescu M, et al. Impact of hepatitis C virus core mutations on the response to interferon-based treatment in chronic hepatitis C. World J Gastroenterol. 2016;22(37):8406-13. DOI: 10.3748/wjg.v22.i37.8406

18. Dinu S, Calistru PI, Ceausu E, Tardei G, Oprisan G. Screening of Protease Inhibitors Resistance Mutations in Hepatitis C Virus Isolates Infecting Romanian Patients Unexposed to Triple Therapy. Roum Arch Microbiol Immunol. 2015;74(1-2):7-17.

19. Hall TA. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser. 1999;41:95-8.

20. Kalaghatgi P, Sikorski AM, Knops E, Rupp D, Sierra S, Heger E, et al. Geno2pheno[HCV] - A Web-based Interpretation System to Support Hepatitis C Treatment Decisions in the Era of Direct-Acting Antiviral Agents. PloS One. 2016;11(5):e0155869. DOI: 10.1371/journal.pone.0155869

21. Asselah T, Marcellin P, Schinazi RF. Treatment of hepatitis C virus infection with direct-acting antiviral agents: 100% cure? Liver Int. 2018;38 Suppl 1:7-13. DOI: 10.1111/liv.13673

22. Bagaglio S, Uberti-Foppa C, Morsica G. Resistance Mechanisms in Hepatitis C Virus: implications for Direct-Acting Antiviral Use. Drugs. 2017;77(10):1043-55. DOI: 10.1007/s40265-017-0753-x

23. European Association for the Study of the Liver. EASL Recommendations on Treatment of Hepatitis C 2018. J Hepatol. 2018 Apr 9. pii:S0168-8278(18)31968-8.

24. Pawlotsky JM. Hepatitis C Virus Resistance to Direct-Acting Antiviral Drugs in Interferon-Free Regimens. Gastroenterology. 2016;151(1):70-86. DOI: 10.1053/j.gastro.2016.04.003

25. Vidal LL, Soares MA, Santos AF. NS3 protease polymorphisms and genetic barrier to drug resistance of distinct hepatitis C virus genotypes from worldwide treatment-naïve subjects. J Viral Hepat. 2016;23(11):840-849. DOI: 10.1111/jvh.12503

26. Kliemann DA, Tovo CV, da Veiga AB, de Mattos AA, Wood C. Polymorphisms and resistance mutations of hepatitis C virus on sequences in the European hepatitis C virus database. World J Gastroenterol. 2016;22(40):8910-7. DOI: 10.3748/wjg.v22.i40.8910

27. Sargin Altunok E, Sayan M, Akhan S, Aygen B, Yildiz O, Tekin Koruk S, et al. Protease Inhibitors Drug Resistance Mutations in Turkish Patients with Chronic Hepatitis C. Int J Infect Dis. 2016;50:1-5. DOI: 10.1016/j.ijid.2016.07.003

28. Echeverria N, Betancour G, Gambaro F, Hernandez N, Lopez P, Chiodi D, et al. Naturally occurring NS3 resistance-associated variants in hepatitis C virus genotype 1: Their relevance for developing countries. Virus Res. 2016;223:140-6. DOI: 10.1016/j.virusres.2016.07.008

29. Wyles DL. Resistance to DAAs: When to Look and When It Matters. Curr HIV/AIDS Rep. 2017;14(6):229-37. DOI: 10.1007/s11904-017-0369-5

30. Chevaliez S, Asselah T. Mechanisms of non-response to antiviral treatment in chronic hepatitis C. Clin Res Hepatol Gastroenterol. 2011;35 Suppl 1:S31-41. DOI: 10.1016/S2210-7401(11)70005-5

31. Chen ZW, Li H, Ren H, Hu P. Global prevalence of pre-existing HCV variants resistant to direct-acting antiviral agents (DAAs): mining the GenBank HCV genome data. Sci Rep. 2016;6:20310. DOI: 10.1038/srep20310

32. Love RA, Parge HE, Wickersham JA, Hostomsky Z, Habuka N, Moomaw EW, et al. The crystal structure of hepatitis C virus NS3 proteinase reveals a trypsin-like fold and a structural zinc binding site. Cell. 1996;87(2):331-42. DOI: 10.1016/S0092-8674(00)81350-1

33. Stempniak M, Hostomska Z, Nodes BR, Hostomsky Z. The NS3 proteinase domain of hepatitis C virus is a zinc-containing enzyme. J Virol. 1997;71(4):2881-6.

34. Dietz J, Susser S, Berkowski C, Perner D, Zeuzem S, Sarrazin C. Consideration of Viral Resistance for Optimization of Direct Antiviral Therapy of Hepatitis C Virus Genotype 1-Infected Patients. PloS One. 2015;10(8):e0134395. DOI: 10.1371/journal.pone.0134395

35. Zhou K, Liang Z, Wang C, Hu F, Ning C, Lan Y, et al. Natural Polymorphisms Conferring Resistance to HCV Protease and Polymerase Inhibitors in Treatment-Naive HIV/HCV Co-Infected Patients in China. PloS One. 2016;11(6):e0157438. DOI: 10.1371/journal.pone.0157438

36. Dietz J, Susser S, Vermehren J, Peiffer KH, Grammatikos G, Berger A, et al. Patterns of Resistance-associated Substitutions in Patients With Chronic HCV Infection Following Treatment With Direct-acting Antivirals. Gastroenterology. 2017;154(4):976-988.e4. DOI: 10.1053/j.gastro.2017.11.007

37. Iio E, Shimada N, Abe H, Atsukawa M, Yoshizawa K, Takaguchi K, et al. Efficacy of daclatasvir/asunaprevir according to resistance-associated variants in chronic hepatitis C with genotype 1. J Gastroenterol. 2017;52(1):94-103. DOI: 10.1007/s00535-016-1225-x

38. Susser S, Welsch C, Wang Y, Zettler M, Domingues FS, Karey U, et al. Characterization of resistance to the protease inhibitor boceprevir in hepatitis C virus-infected patients. Hepatology. 2009;50(6):1709-18. DOI: 10.1002/hep.23192

39. Shepherd SJ, Abdelrahman T, MacLean AR, Thomson EC, Aitken C, Gunson RN. Prevalence of HCV NS3 pre-treatment resistance associated amino acid variants within a Scottish cohort. J Clin Virol. 2015;65:50-3. DOI: 10.1016/j.jcv.2015.02.005

40. Barnard RJ, Howe JA, Ogert RA, Zeuzem S, Poordad F, Gordon SC, et al. Analysis of boceprevir resistance associated amino acid variants (RAVs) in two phase 3 boceprevir clinical studies. Virology. 2013;444(1-2):329-36. DOI: 10.1016/j.virol.2013.06.029

41. Fourati S, Pawlotsky JM. Virologic Tools for HCV Drug Resistance Testing. Viruses. 2015;7(12):6346-59. DOI: 10.3390/v7122941

42. Thomson E, Ip CL, Badhan A, Christiansen MT, Adamson W, Ansari MA, et al. Comparison of Next-Generation Sequencing Technologies for Comprehensive Assessment of Full-Length Hepatitis C Viral Genomes. J Clin Microbiol. 2016;54(10):2470-84. DOI: 10.1128/JCM.00330-16

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Romanian Journal of Laboratory Medicine

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