Department of Systems Biology, Institute of Cytology and Genetics SB RAS, Novosibirsk, Russia.
Department of Computational Biology, St. Petersburg State Polytechnical University, St. Petersburg, Russia.
PLoS One. 2014 Mar 18;9(3):e91502. doi: 10.1371/journal.pone.0091502. eCollection 2014.
As an RNA virus, hepatitis C virus (HCV) is able to rapidly acquire drug resistance, and for this reason the design of effective anti-HCV drugs is a real challenge. The HCV subgenomic replicon-containing cells are widely used for experimental studies of the HCV genome replication mechanisms, for drug testing in vitro and in studies of HCV drug resistance. The NS3/4A protease is essential for virus replication and, therefore, it is one of the most attractive targets for developing specific antiviral agents against HCV. We have developed a stochastic model of subgenomic HCV replicon replication, in which the emergence and selection of drug resistant mutant viral RNAs in replicon cells is taken into account. Incorporation into the model of key NS3 protease mutations leading to resistance to BILN-2061 (A156T, D168V, R155Q), VX-950 (A156S, A156T, T54A) and SCH 503034 (A156T, A156S, T54A) inhibitors allows us to describe the long term dynamics of the viral RNA suppression for various inhibitor concentrations. We theoretically showed that the observable difference between the viral RNA kinetics for different inhibitor concentrations can be explained by differences in the replication rate and inhibitor sensitivity of the mutant RNAs. The pre-existing mutants of the NS3 protease contribute more significantly to appearance of new resistant mutants during treatment with inhibitors than wild-type replicon. The model can be used to interpret the results of anti-HCV drug testing on replicon systems, as well as to estimate the efficacy of potential drugs and predict optimal schemes of their usage.
作为一种 RNA 病毒,丙型肝炎病毒(HCV)能够迅速获得耐药性,因此设计有效的抗 HCV 药物是一个真正的挑战。含有 HCV 亚基因组复制子的细胞广泛用于 HCV 基因组复制机制的实验研究、体外药物测试以及 HCV 耐药性研究。NS3/4A 蛋白酶对于病毒复制是必需的,因此它是开发针对 HCV 的特异性抗病毒药物的最有吸引力的靶标之一。我们已经开发了一种 HCV 亚基因组复制子复制的随机模型,其中考虑了耐药突变病毒 RNA 在复制子细胞中的出现和选择。将导致对 BILN-2061(A156T、D168V、R155Q)、VX-950(A156S、A156T、T54A)和 SCH 503034(A156T、A156S、T54A)抑制剂耐药的关键 NS3 蛋白酶突变纳入模型,使我们能够描述各种抑制剂浓度下病毒 RNA 抑制的长期动力学。我们从理论上表明,不同抑制剂浓度下病毒 RNA 动力学的可观察差异可以用突变 RNA 的复制率和抑制剂敏感性的差异来解释。在抑制剂治疗过程中,NS3 蛋白酶的预先存在的突变体比野生型复制子对新的耐药突变体的出现贡献更大。该模型可用于解释在复制子系统上进行抗 HCV 药物测试的结果,以及估计潜在药物的疗效并预测其使用的最佳方案。