Protease inhibitor-resistant hepatitis C virus mutants with reduced fitness from impaired production of infectious virus

Tetsuro Shimakami, Christoph Welsch, Daisuke Yamane, David R. McGivern, Min Kyung Yi, Stefan Zeuzem, Stanley M. Lemon

Research output: Contribution to journalArticle

107 Citations (Scopus)

Abstract

Background & Aims Several small molecule inhibitors of the hepatitis C virus (HCV) nonstructural protein (NS) 3/4A protease have advanced successfully to clinical trials. However, the selection of drug-resistant mutants is a significant issue with protease inhibitors (PIs). A variety of amino acid substitutions in the protease domain of NS3 can lead to PI resistance. Many of these significantly impair the replication fitness of HCV RNA replicons. However, it is not known whether these mutations also adversely affect infectious virus assembly and release, processes in which NS3 also participates. Methods We studied the impact of 25 previously identified PI-resistance mutations on the capacity of genotype 1a H77S RNA to replicate in cell culture and produce infectious virus. Results Most PI-resistance mutations resulted in moderate loss of replication competence, although several (V36A/L/M, R109K, and D168E) showed fitness comparable to wild type, whereas others (S138T and A156V) were severely impaired both in RNA replication and infectious virus production. Although reductions in RNA replication capacity correlated with decreased yields of infectious virus for most mutations, a subset of mutants (Q41R, F43S, R155T, A156S, and I170A/T) showed greater impairment in their ability to produce virus than predicted from reductions in RNA replication capacity. Detailed examination of the I170A mutant showed no defect in release of virus from cells and no significant difference in specific infectivity of extracellular virus particles. Conclusions Replicon-based assays might underestimate the loss of fitness caused by PI-resistance mutations, because some mutations in the NS3 protease domain specifically impair late steps in the viral life cycle that involve intracellular assembly of infectious virus.

Original languageEnglish (US)
Pages (from-to)667-675
Number of pages9
JournalGastroenterology
Volume140
Issue number2
DOIs
StatePublished - Feb 2011

Fingerprint

Protease Inhibitors
Hepacivirus
Viruses
Mutation
RNA
Virus Release
Virus Assembly
Replicon
Peptide Hydrolases
Aptitude
Amino Acid Substitution
Virus Replication
Life Cycle Stages
Virion
Mental Competency
Cell Culture Techniques
Genotype
Clinical Trials
Pharmaceutical Preparations
Proteins

Keywords

  • Antiviral Drug
  • NS3/4A Protease
  • Viral Assembly
  • Viral Replication

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Protease inhibitor-resistant hepatitis C virus mutants with reduced fitness from impaired production of infectious virus. / Shimakami, Tetsuro; Welsch, Christoph; Yamane, Daisuke; McGivern, David R.; Yi, Min Kyung; Zeuzem, Stefan; Lemon, Stanley M.

In: Gastroenterology, Vol. 140, No. 2, 02.2011, p. 667-675.

Research output: Contribution to journalArticle

Shimakami, Tetsuro ; Welsch, Christoph ; Yamane, Daisuke ; McGivern, David R. ; Yi, Min Kyung ; Zeuzem, Stefan ; Lemon, Stanley M. / Protease inhibitor-resistant hepatitis C virus mutants with reduced fitness from impaired production of infectious virus. In: Gastroenterology. 2011 ; Vol. 140, No. 2. pp. 667-675.
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