Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, Kentucky, USA.
J Virol. 2013 Jun;87(12):6866-75. doi: 10.1128/JVI.03475-12. Epub 2013 Apr 10.
Our recent studies demonstrated that apolipoprotein E mediates cell attachment of hepatitis C virus (HCV) through interactions with the cell surface heparan sulfate (HS). HS is known to covalently attach to core proteins to form heparan sulfate proteoglycans (HSPGs) on the cell surface. The HSPG core proteins include the membrane-spanning syndecans (SDCs), the lycosylphosphatidylinositol-linked glypicans (GPCs), the basement membrane proteoglycan perlecan (HSPG2), and agrin. In the present study, we have profiled each of the HSPG core proteins in HCV attachment. Substantial evidence derived from our studies demonstrates that SDC1 is the major receptor protein for HCV attachment. The knockdown of SDC1 expression by small interfering RNA (siRNA)-induced gene silence resulted in a significant reduction of HCV attachment to Huh-7.5 cells and stem cell-differentiated human hepatocytes. The silence of SDC2 expression also caused a modest decrease of HCV attachment. In contrast, the siRNA-mediated knockdown of other SDCs, GPCs, HSPG2, and agrin had no effect on HCV attachment. More importantly, ectopic expression of SDC1 was able to completely restore HCV attachment to Huh-7.5 cells in which the endogenous SDC1 expression was silenced by specific siRNAs. Interestingly, mouse SDC1 is also fully functional in mediating HCV attachment when expressed in the SDC1-deficient cells, consistent with recent reports that mouse hepatocytes are also susceptible to HCV infection when expressing other key HCV receptors. Collectively, our findings demonstrate that SDC1 serves as the major receptor protein for HCV attachment to cells, providing another potential target for discovery and development of antiviral drugs against HCV.
我们最近的研究表明,载脂蛋白 E 通过与细胞表面硫酸乙酰肝素 (HS) 的相互作用介导丙型肝炎病毒 (HCV) 的细胞附着。HS 已知与核心蛋白共价结合,在细胞表面形成硫酸乙酰肝素蛋白聚糖 (HSPGs)。HSPG 核心蛋白包括跨膜连接蛋白聚糖 (SDC)、糖基磷脂酰肌醇连接的聚糖 (GPC)、基底膜蛋白聚糖 perlecan (HSPG2) 和 agrin。在本研究中,我们对 HCV 附着的每种 HSPG 核心蛋白进行了分析。我们的研究得出了大量证据,证明 SDC1 是 HCV 附着的主要受体蛋白。通过小干扰 RNA (siRNA) 诱导的基因沉默敲低 SDC1 表达导致 HCV 对 Huh-7.5 细胞和干细胞分化的人肝细胞的附着显著减少。SDC2 表达的沉默也导致 HCV 附着适度减少。相比之下,siRNA 介导的其他 SDC、GPC、HSPG2 和 agrin 的沉默对 HCV 附着没有影响。更重要的是,SDC1 的异位表达能够完全恢复 Huh-7.5 细胞中的 HCV 附着,而这些细胞中的内源性 SDC1 表达被特异性 siRNA 沉默。有趣的是,当在 SDC1 缺陷细胞中表达时,鼠 SDC1 也完全能够介导 HCV 附着,这与最近的报道一致,即当表达其他关键 HCV 受体时,鼠肝细胞也容易感染 HCV。总之,我们的发现表明 SDC1 是 HCV 附着到细胞的主要受体蛋白,为发现和开发抗 HCV 抗病毒药物提供了另一个潜在靶点。