Evolution of resistant M414T mutants among hepatitis C virus replicon cells treated with polymerase inhibitor A-782759.
Treatment of hepatitis C virus (HCV) replicon cells with any single specific anti-HCV inhibitor in vitro leads to a rapid selection of resistant mutants. However, the source and the kinetic evolution of these resistant mutants during treatment are poorly understood. In this study we developed allele-specific real-time PCR assays for quantitative detection of the M414T mutant that was selected by a number of benzothiadiazine HCV polymerase inhibitors. Low levels of preexisting M414T mutants were detected in both 1b-con1 (0.22%) and 1b-N (0.18%) subgenomic replicon cell lines, as well as in 6 of 15 HCV RNA isolated from the sera of treatment-naive HCV-infected patients ranging from 0.11 to 0.60%. The proportion of M414T mutants in replicons rapidly increased in a dose-dependent manner upon treatment with benzothiadiazine inhibitor A-782759. After 4 days of treatment, 2.5, 26, or 60% of the replicon population contained M414T mutants with the use of A-782759 at 1x, 10x, or 100x its 50% effective concentration, respectively. In addition, the short 4-day treatment resulted in significant changes in inhibitor susceptibility in the replicon cells. Our results indicated that the resistant mutant preexisted as a minor population in replicon cells and that the mutant was selected within days of treatment with the inhibitor. The findings from this study suggested that early application of combination therapy of an HCV-specific inhibitor with interferon-based regimens or other classes of available inhibitors will be necessary to avoid quick viral rebound or treatment failure.
Lu
Liangjun
L
Global Pharmaceutical Research and Development, Abbott Laboratories, GPRD-R4CQ, 200 Abbott Park Rd., Abbott Park, IL 60064, USA. liangjun.lu@abbott.com
Mo
Hongmei
H
Pilot-Matias
Tami J
TJ
Molla
Akhteruzzaman
A
D001581:Benzothiadiazines / Q000494:pharmacology; D024882:Drug Resistance, Viral / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019143:Evolution, Molecular; D005838:Genotype; D016174:Hepacivirus / Q000145:classification / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D006526:Hepatitis C / Q000821:virology; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D012324:RNA-Dependent RNA Polymerase / Q000037:antagonists & inhibitors* / Q000235:genetics; D012093:Replicon / Q000187:drug effects / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics
D001581:Benzothiadiazines; D004791:Enzyme Inhibitors; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase