1887

Abstract

Hepatitis C virus (HCV) genotype 3 presents a high level of both baseline and acquired resistance to direct-acting antivirals (DAAs), particularly those targeting the NS5A protein. To understand this resistance we studied a cohort of Brazilian patients treated with the NS5A DAA, daclatasvir and the nucleoside analogue, sofosbuvir. We observed a novel substitution at NS5A amino acid residue 98 [serine to glycine (S98G)] in patients who relapsed post-treatment. The effect of this substitution on both replication fitness and resistance to DAAs was evaluated using two genotype 3 subgenomic replicons. S98G had a modest effect on replication, but in combination with the previously characterized resistance-associated substitution (RAS), Y93H, resulted in a significant increase in daclatasvir resistance. This result suggests that combinations of substitutions may drive a high level of DAA resistance and provide some clues to the mechanism of action of the NS5A-targeting DAAs.

Funding
This study was supported by the:
  • FAPESP (Award 2018/04678-5)
    • Principle Award Recipient: Guilherme Rodrigues Fernandes Campos
  • FAPESP (Award 2016/03807-0)
    • Principle Award Recipient: Guilherme Rodrigues Fernandes Campos
  • China Scholarship Council, http://dx.doi.org/10.13039/501100004543
  • Medical Research Council, http://dx.doi.org/10.13039/501100000265 (Award MR/S001026/1)
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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/content/journal/jgv/10.1099/jgv.0.001496
2020-11-03
2024-04-25
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References

  1. Smith DB, Bukh J, Kuiken C, Muerhoff AS, Rice CM et al. Expanded classification of hepatitis C virus into 7 genotypes and 67 subtypes: updated criteria and genotype assignment web resource. Hepatology 2014; 59:318–327 [View Article] [PubMed]
    [Google Scholar]
  2. Chan A, Patel K, Naggie S. Genotype 3 infection: the last stand of hepatitis C virus. Drugs 2017; 77:131–144 [View Article] [PubMed]
    [Google Scholar]
  3. Kattakuzhy S, Levy R, Rosenthal E, Tang L, Wilson E et al. Hepatitis C genotype 3 disease. Hepatol Int 2016; 10:861–870 [View Article] [PubMed]
    [Google Scholar]
  4. Nutini MFR, Hunter J, Giron L, Pires A, Kohiyama IM et al. HCV genotype profile in Brazil of mono-infected and HIV co-infected individuals: a survey representative of an entire country. PLoS One 2020; 15:e0227082 [View Article] [PubMed]
    [Google Scholar]
  5. Gao M, Nettles RE, Belema M, Snyder LB, Nguyen VN et al. Chemical genetics strategy identifies an HCV NS5A inhibitor with a potent clinical effect. Nature 2010; 465:96–100 [View Article] [PubMed]
    [Google Scholar]
  6. Kelly L, Badhan A, Roberts GC, Mbisa JL, Harris M. Manipulation of both virus- and cell-specific factors is required for robust transient replication of a hepatitis C virus genotype 3A sub-genomic replicon. J Gen Virol 2017; 98:2495–2506 [View Article] [PubMed]
    [Google Scholar]
  7. Saeed M, Scheel TKH, Gottwein JM, Marukian S, Dustin LB et al. Efficient replication of genotype 3A and 4A hepatitis C virus replicons in human hepatoma cells. Antimicrob Agents Chemother 2012; 56:5365–5373 [View Article] [PubMed]
    [Google Scholar]
  8. Ward JC, Bowyer S, Chen S, Fernandes Campos GR, Ramirez S et al. Insights into the unique characteristics of hepatitis C virus genotype 3 revealed by development of a robust sub-genomic DBN3a replicon. J Gen Virol 2020 08 Sep 2020 [View Article] [PubMed]
    [Google Scholar]
  9. Ramirez S, Mikkelsen LS, Gottwein JM, Bukh J. Robust HCV genotype 3A infectious cell culture system permits identification of escape variants with resistance to sofosbuvir. Gastroenterology 2016; 151:e972973–985 [View Article] [PubMed]
    [Google Scholar]
  10. Witteveldt J, Martin-Gans M, Simmonds P. Enhancement of the replication of hepatitis C virus replicons of genotypes 1 to 4 by manipulation of CpG and uPA dinucleotide frequencies and use of cell lines expressing SECL14L2 for antiviral resistance testing. Antimicrob Agents Chemother 2016; 60:2981–2992 [View Article] [PubMed]
    [Google Scholar]
  11. Nettles RE, Gao M, Bifano M, Chung E, Persson A et al. Multiple ascending dose study of BMS-790052, a nonstructural protein 5A replication complex inhibitor, in patients infected with hepatitis C virus genotype 1. Hepatology 2011; 54:1956–1965 [View Article] [PubMed]
    [Google Scholar]
  12. Ke R, Loverdo C, Qi H, Olson CA, Wu NC et al. Modelling clinical data shows active tissue concentration of daclatasvir is 10-fold lower than its plasma concentration. J Antimicrob Chemother 2014; 69:724–727 [View Article] [PubMed]
    [Google Scholar]
  13. Tellinghuisen TL, Marcotrigiano J, Rice CM. Structure of the zinc-binding domain of an essential component of the hepatitis C virus replicase. Nature 2005; 435:374–379 [View Article] [PubMed]
    [Google Scholar]
  14. Love RA, Brodsky O, Hickey MJ, Wells PA, Cronin CN. Crystal structure of a novel dimeric form of NS5A domain I protein from hepatitis C virus. J Virol 2009; 83:4395–4403 [View Article] [PubMed]
    [Google Scholar]
  15. Yin C, Goonawardane N, Stewart H, Harris M. A role for domain I of the hepatitis C virus NS5A protein in virus assembly. PLoS Pathog 2018; 14:e1006834 [View Article] [PubMed]
    [Google Scholar]
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