Mutations in the chikungunya virus non-structural proteins cause resistance to favipiravir (T-705), a broad-spectrum antiviral
Mutations in the chikungunya virus non-structural proteins cause resistance to favipiravir (T-705), a broad-spectrum antiviral
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DOI：
10.1093/jac/dku209
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发表时间：
2014-10-01
影响因子：
5.2
通讯作者：
Leyssen, Pieter
中科院分区：
文献类型：
--
作者：
Delang, Leen;Segura Guerrero, Nidya;Leyssen, Pieter
目的：T-705，也称为法匹拉韦，是一种小分子抑制剂，目前正在临床开发中，用于治疗流感病毒感染。这种分子还抑制了其他RNA病毒的广泛复制。本研究的目的是探讨法匹拉韦对基孔肯雅病毒（CHIKV）复制的抗病毒作用，并有助于解开这种病毒的分子作用机制。方法：在细胞培养和小鼠致死性感染模型中检测法匹拉韦的抗CHIKV作用。使用五步方案来选择对法匹拉韦敏感性降低的CHIKV变体。在细胞培养中证实了抗性表型，并进行了全基因组测序。将鉴定的突变反向工程改造成感染性克隆，以确认其对法匹拉韦抗病毒疗效的影响。结果：法匹拉韦抑制CHIKV的实验室菌株和临床分离株以及一组其他甲病毒的复制。独立选择了几种法匹拉韦耐药CHIKV变体，所有这些变体都特别获得了RNA依赖性RNA聚合酶（RdRp）中独特的K291 R突变。将这种K291 R突变反向工程化到CHIKV的感染性克隆中证实了突变基因型和抗性表型之间的联系。有趣的是，这种特殊的赖氨酸在一般正链RNA病毒的RdRp中也是高度保守的。
Objectives: T-705, also known as favipiravir, is a small-molecule inhibitor that is currently in clinical development for the treatment of influenza virus infections. This molecule also inhibits the replication of a broad spectrum of other RNA viruses. The objective of this study was to investigate the antiviral effect of favipiravir on chikungunya virus (CHIKV) replication and to contribute to unravelling the molecular mechanism of action against this virus.Methods: The anti-CHIKV effect of favipiravir was examined in cell culture and in a mouse model of lethal infection. A five-step protocol was used to select for CHIKV variants with reduced susceptibility to favipiravir. The resistant phenotype was confirmed in cell culture and the whole genome was sequenced. The identified mutations were reverse-engineered into an infectious clone to confirm their impact on the antiviral efficacy of favipiravir.Results: Favipiravir inhibits the replication of laboratory strains and clinical isolates of CHIKV, as well as of a panel of other alphaviruses. Several favipiravir-resistant CHIKV variants were independently selected and all of them in particular acquired the unique K291R mutation in the RNA-dependent RNA polymerase (RdRp). Reverse-engineering of this K291R mutation into an infectious clone of CHIKV confirmed the link between the mutant genotype and the resistant phenotype. Interestingly, this particular lysine is also highly conserved in the RdRp of positivestranded RNA viruses in general.Conclusions: This study provides an important insight into the precise molecular mechanism by which favipiravir exerts its