Tao Wanyin, Gan Tianyu, Lu Jie, Zhong Jin
Unit of Viral Hepatitis, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, PR China.
Unit of Viral Hepatitis, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, PR China; University of Chinese Academy of Sciences, Beijing, 100049, PR China.
Antiviral Res. 2017 Mar;139:18-24. doi: 10.1016/j.antiviral.2016.12.009. Epub 2016 Dec 23.
The development of direct-acting antivirals (DAAs) has significantly improved hepatitis C virus (HCV) treatment. However, drug resistance remains a potential concern in the real-world DAA-based therapies. We previously developed a novel full-length genotype 2a HCVcc clone PR63cc directly from clinical isolates. Here in this study, we compared the sensitivity of PR63cc and JFH1 to 12 different DAAs most of which are either already in clinical use or in the late clinical development phase. For NS5B inhibitors, PR63cc and JFH1 displayed comparable sensitivity to nucleoside/nucleotide analogues sofosbuvir and 2'-C-methyladenosine, while PR63cc was 4-fold more sensitive than JFH1 to nesbuvir, a non-nucleoside inhibitor. Interestingly, PR63cc and JFH1 were both completely resistant to dasabuvir which efficiently inhibited the replication of genotype 1b HCV replicon. For NS5A inhibitors, while PR63cc was as sensitive as JFH1 to ombitasvir and velpatasvir, it was much more resistant than JFH1 to daclatasvir and ledipasvir, which was mainly due to methionine at amino acid residue 31 of NS5A. For NS3 inhibitors, PR63cc was generally less sensitive than JFH1 to simeprevir, grazoprevir, asunaprevir and paritaprevir. Serine at residue 67 of NS3 was identified to be a resistance-associated variant (RAV) for asunaprevir. Finally, we showed that PR63cc was more resistant than JFH1 to the asunaprevir/daclatasvir combination treatment. In summary, our study systemically analyzed the DAA sensitivity of a new HCVcc strain and identified critical RAVs. These results are not only important for monitoring the emergence of drug-resistant mutations of current DAA therapies, but also valuable for developing next-generation DAAs.
直接作用抗病毒药物（DAA）的发展显著改善了丙型肝炎病毒（HCV）的治疗。然而，在基于DAA的实际治疗中，耐药性仍然是一个潜在问题。我们之前直接从临床分离株中开发出一种新型的全长2a基因型HCVcc克隆PR63cc。在本研究中，我们比较了PR63cc和JFH1对12种不同DAA的敏感性，其中大多数已在临床使用或处于临床开发后期阶段。对于NS5B抑制剂，PR63cc和JFH1对核苷/核苷酸类似物索磷布韦和2'-C-甲基腺苷表现出相当的敏感性，而PR63cc对非核苷抑制剂奈拉布韦的敏感性比JFH1高4倍。有趣的是，PR63cc和JFH1对能有效抑制1b基因型HCV复制子复制的达沙布韦均完全耐药。对于NS5A抑制剂，虽然PR63cc对奥比他韦和维帕他韦的敏感性与JFH1相当，但它对达卡他韦和来迪帕司韦的耐药性比JFH1强得多，这主要是由于NS5A氨基酸残基31处的甲硫氨酸所致。对于NS3抑制剂，PR63cc对simeprevir、grazoprevir、asunaprevir和paritaprevir的敏感性通常低于JFH1。NS3残基67处的丝氨酸被确定为asunaprevir的耐药相关变异（RAV）。最后，我们表明PR63cc对asunaprevir/达卡他韦联合治疗的耐药性比JFH1更强。总之，我们的研究系统地分析了一种新的HCVcc毒株对DAA的敏感性，并确定了关键的RAV。这些结果不仅对于监测当前DAA治疗中耐药突变的出现很重要，而且对于开发下一代DAA也很有价值。
