National Institute of Biomedical Innovation, 7-6-8 Saito Asagi, Ibaraki, Osaka, 567-0085, Japan.
J Proteome Res. 2012 Jul 6;11(7):3664-79. doi: 10.1021/pr300121a. Epub 2012 Jun 12.
Hepatitis C virus (HCV) causes chronic liver disease worldwide. HCV Core protein (Core) forms the viral capsid and is crucial for HCV pathogenesis and HCV-induced hepatocellular carcinoma, through its interaction with the host factor proteasome activator PA28γ. Here, using BD-PowerBlot high-throughput Western array, we attempt to further investigate HCV pathogenesis by comparing the protein levels in liver samples from Core-transgenic mice with or without the knockout of PA28γ expression (abbreviated PA28γ(-/-)CoreTG and CoreTG, respectively) against the wild-type (WT). The differentially expressed proteins integrated into the human interactome were shown to participate in compact and well-connected cellular networks. Functional analysis of the interaction networks using a newly developed data warehouse system highlighted cellular pathways associated with vesicular transport, immune system, cellular adhesion, and cell growth and death among others that were prominently influenced by Core and PA28γ in HCV infection. Follow-up assays with in vitro HCV cell culture systems validated VTI1A, a vesicular transport associated factor, which was upregulated in CoreTG but not in PA28γ(-/-)CoreTG, as a novel regulator of HCV release but not replication. Our analysis provided novel insights into the Core-PA28γ interplay in HCV pathogenesis and identified potential targets for better anti-HCV therapy and potentially novel biomarkers of HCV infection.
丙型肝炎病毒（HCV）在全球范围内引起慢性肝病。HCV 核心蛋白（Core）形成病毒衣壳，通过与宿主因子蛋白酶体激活剂 PA28γ 的相互作用，对 HCV 发病机制和 HCV 诱导的肝细胞癌至关重要。在这里，我们使用 BD-PowerBlot 高通量 Western 阵列，通过比较表达 PA28γ 敲除（简称 PA28γ(-/-)CoreTG 和 CoreTG）或不表达 PA28γ 的 Core 转基因小鼠的肝组织样本中的蛋白质水平，尝试进一步研究 HCV 的发病机制，与野生型（WT）相比。整合到人类相互作用组中的差异表达蛋白参与了紧密且连接良好的细胞网络。使用新开发的数据仓库系统对相互作用网络进行功能分析，突出了与囊泡运输、免疫系统、细胞黏附以及细胞生长和死亡等相关的细胞途径，这些途径在 HCV 感染中受到 Core 和 PA28γ 的显著影响。体外 HCV 细胞培养系统的后续检测验证了 VTI1A，一种囊泡运输相关因子，在 CoreTG 中上调，但在 PA28γ(-/-)CoreTG 中没有上调，作为 HCV 释放但不是复制的新型调节剂。我们的分析为 HCV 发病机制中的 Core-PA28γ 相互作用提供了新的见解，并确定了更好的抗 HCV 治疗和 HCV 感染的潜在新型生物标志物的潜在靶点。
