A cell culture system for distinguishing hepatitis C viruses with and without liver cancer-related mutations in the viral core gene.
A cell culture system for distinguishing hepatitis C viruses with and without liver cancer-related mutations in the viral core gene.
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DOI：
10.1016/j.jhep.2015.07.024
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发表时间：
2015-12
影响因子：
25.7
通讯作者：
Branch AD
中科院分区：
文献类型：
--
作者：
El-Shamy A;Eng FJ;Doyle EH;Klepper AL;Sun X;Sangiovanni A;Iavarone M;Colombo M;Schwartz RE;Hoshida Y;Branch AD
虽然感染带有Q70和/或M91核心基因突变的1b型丙型肝炎病毒的患者患肝细胞癌的风险增加了近五倍，并增加了胰岛素抵抗，但由于缺乏合适的实验系统，无法直接实验这些突变对细胞基因表达的影响。用人血清长期处理HH-7细胞以诱导分化，并建立检测高危和控制丙型肝炎病毒的模型系统。为了临床验证，将感染细胞的轮廓相互比较，并与前瞻性随访长达23年的早期丙型肝炎相关肝硬变患者的肝活检进行比较(n=216)。在人血清中长期培养产生生长停滞的肝细胞样细胞，其基因图谱与原代人类肝细胞显著重叠。高危(Q70/M91)和对照(R70/L91)病毒对这些细胞基因表达的影响有显著差异。高危病毒增强了与癌症和II型糖尿病相关的通路的表达，而对照病毒增强了与氧化磷酸化相关的通路。具有特殊临床相关性的是，复制高危病毒的细胞的转录组与患者的肝癌高危特征显著相关(Bonferroni校正P=0.03)，而在非肝癌相关的临床结果中没有观察到这种关联。这种基于细胞的系统允许对丙型肝炎病毒变异进行直接的面对面比较，并为先前的临床数据提供了实验支持，这些数据表明核心基因突变具有致癌作用。这个简单的实验系统区分了丙型肝炎病毒的变种，并将使未来的机制分析和干预方法的探索成为可能。
Although patients infected by genotype-1b hepatitis C virus (HCV) with Q70 and/or M91 core gene mutations have an almost five-fold increased risk of developing hepatocellular carcinoma (HCC) and increased insulin resistance, the absence of a suitable experimental system has precluded direct experimentation on the effects of these mutations on cellular gene expression. Huh-7 cells were treated long-term with human serum to induce differentiation and to produce a model system for testing high-risk and control HCV. For clinical validation, profiles of infected cells were compared to each other and to those of liver biopsies of patients with early-stage HCV-related cirrhosis followed prospectively for up to 23 years (n=216). Long-term culture in human serum produced growth-arrested, hepatocyte-like cells whose gene profile overlapped significantly with that of primary human hepatocytes. High-risk (Q70/M91) and control (R70/L91) viruses had dramatically different effects on gene expression of these cells. The high-risk virus enhanced expression of pathways associated with cancer and type II diabetes, while the control virus enhanced pathways associated with oxidative phosphorylation. Of special clinical relevance, the transcriptome of cells replicating the high-risk virus correlated significantly with an HCC high-risk profile in patients (Bonferroni-corrected P=0.03), whereas no such association was observed for non-HCC-related clinical outcomes. The cell-based system allowed direct head-to-head comparison of HCV variants and provided experimental support for previous clinical data indicating an oncogenic effect of core gene mutations. This simple experimental system distinguished HCV variants and will enable future mechanistic analysis and exploration of interventional approaches.