Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9048, USA.
J Virol. 2011 Jul;85(14):7273-83. doi: 10.1128/JVI.00629-11. Epub 2011 May 4.
Many individuals infected with hepatitis C virus (HCV) develop a chronic infection, and of those who are treated with pegylated interferon and ribavirin (RBV), many do not respond. While the nucleoside analog RBV improves treatment outcome, and will likely be an important component of therapy with next-generation viral inhibitors, RBV's mechanism is controversial. Most of RBV's proposed mechanisms require RBV import into cells. Therefore, we explored whether host-based RBV resistance develops through reduced cellular uptake, akin to chemotherapy resistance in some cancers. We examined the effect of host-based RBV resistance on HCV replication in cultured hepatoma Huh7.5 liver cells and whether RBV resistance develops in HCV patients. When Huh7.5 cells were exposed to RBV, resistance developed through reduced RBV uptake via the ENT1 nucleoside transporter and antiviral efficacy was reduced. The uptake defect in RBV-resistant cells was specific to RBV, since transport of another ENT1 substrate, cytidine, was unaffected. Importantly, RBV uptake significantly declined in HCV patient peripheral blood mononuclear cells (PBMCs) following 4 weeks of therapy. Furthermore, maintenance of RBV uptake correlated with rapid treatment response. Our results uncovered a novel form of antiviral drug resistance and suggest that host-based RBV resistance develops in HCV patients undergoing therapy and that maintenance of RBV uptake may contribute to rapid viral clearance.
许多感染丙型肝炎病毒 (HCV) 的个体都会发展成慢性感染,而在接受聚乙二醇干扰素和利巴韦林 (RBV) 治疗的患者中,许多人没有应答。虽然核苷类似物 RBV 改善了治疗效果,并且可能成为下一代病毒抑制剂治疗的重要组成部分,但 RBV 的作用机制仍存在争议。RBV 的大多数提议的机制需要 RBV 进入细胞。因此,我们探讨了宿主 RBV 耐药性是否通过减少细胞摄取而发展,类似于某些癌症中的化疗耐药性。我们研究了宿主 RBV 耐药性对培养的肝癌 Huh7.5 肝细胞中 HCV 复制的影响,以及 HCV 患者中是否会出现 RBV 耐药性。当 Huh7.5 细胞暴露于 RBV 时,通过 ENT1 核苷转运蛋白减少 RBV 摄取而产生耐药性,抗病毒功效降低。RBV 耐药细胞中的摄取缺陷是 RBV 特异性的,因为另一种 ENT1 底物胞苷的转运不受影响。重要的是,在接受治疗的 4 周后,HCV 患者外周血单核细胞 (PBMC) 中的 RBV 摄取显著下降。此外,RBV 摄取的维持与快速治疗应答相关。我们的结果揭示了一种新的抗病毒药物耐药形式,并表明在接受治疗的 HCV 患者中会出现宿主 RBV 耐药性,并且维持 RBV 摄取可能有助于快速清除病毒。