Green Center for Reproductive Biology Sciences, Department of Obstetrics and Gynecology, University of Texas Southwestern Medical School, Dallas, Texas, United States of America.
Pharmacology and Neurosience, University of North Texas Health Science Center, Fort Worth, Texas, United States of America.
PLoS One. 2018 Aug 23;13(8):e0202524. doi: 10.1371/journal.pone.0202524. eCollection 2018.
To elucidate HIV-1 co-infection-induced acceleration of HCV liver disease and identify stage-specific molecular signatures, we applied a new high-resolution molecular screen, the Affymetrix GeneChip Human Transcriptome Array (HTA2.0), to HCV-mono- and HIV/HCV-co-infected liver specimens from subjects with early and advanced disease. Out of 67,528 well-annotated genes, we have analyzed the functional and statistical significance of 75 and 28 genes expressed differentially between early and advanced stages of HCV mono- and HIV/HCV co-infected patient liver samples, respectively. We also evaluated the expression of 25 and 17 genes between early stages of mono- and co-infected liver tissues and between advanced stages of mono- and co-infected patient's samples, respectively. Based on our analysis of fold-change in gene expression as a function of disease stage (i.e., early vs. advanced), coupled with consideration of the known relevant functions of these genes, we focused on four candidate genes, ACSL4, GNMT, IFI27, and miR122, which are expressed stage-specifically in HCV mono- and HIV-1/HCV co-infective liver disease and are known to play a pivotal role in regulating HCV-mediated hepatocellular carcinoma (HCC). Our qRT-PCR analysis of the four genes in patient liver specimens supported the microarray data. Protein products of each gene were detected in the endoplasmic reticulum (ER) where HCV replication takes place, and the genes' expression significantly altered replicability of HCV in the subgenomic replicon harboring regulatory genes of the JFH1 strain of HCV in Huh7.5.1. With respect to three well-known transferrable HIV-1 viral elements-Env, Nef, and Tat-Nef uniquely augmented replicon expression, while Tat, but not the others, substantially modulated expression of the candidate genes in hepatocytic cells. Combinatorial expression of these cellular and viral genes in the replicon cells further altered replicon expression. Taken together, these results showed that HIV-1 viral proteins can exacerbate liver pathology in the co-infected patients by disparate molecular mechanisms-directly or indirectly dysregulating HCV replication, even if lack of association of HCV load and end-stage liver disease in hemophilic patients were reported, and modulating expression of hepatocellular genes critical for disease progression. These findings also provide major insights into development of stage-specific hepatocellular biomarkers for improved diagnosis and prognosis of HCV-mediated liver disease.
为了阐明 HIV-1 合并感染加速 HCV 肝病的机制并确定阶段特异性分子特征，我们应用了一种新的高通量分子筛选方法，即 Affymetrix GeneChip Human Transcriptome Array（HTA2.0），对来自早期和晚期 HCV 单感染和 HIV/HCV 合并感染患者肝脏标本的 HCV 进行分析。在 67528 个注释良好的基因中，我们分析了 HCV 单感染和 HIV/HCV 合并感染患者肝脏样本中早期和晚期之间差异表达的 75 个和 28 个基因的功能和统计学意义。我们还评估了早期 HCV 单感染和合并感染肝组织之间以及晚期 HCV 单感染和合并感染患者样本之间的 25 个和 17 个基因的表达。基于我们对基因表达随疾病阶段变化的分析（即早期与晚期），结合对这些基因已知相关功能的考虑，我们重点关注了 4 个候选基因，即 ACSL4、GNMT、IFI27 和 miR122，它们在 HCV 单感染和 HIV-1/HCV 合并感染性肝病中具有阶段特异性表达，并且已知在调节 HCV 介导的肝细胞癌（HCC）中发挥关键作用。我们对患者肝组织标本中这 4 个基因的 qRT-PCR 分析支持了微阵列数据。每个基因的蛋白质产物都在 HCV 复制发生的内质网（ER）中被检测到，并且这些基因的表达显著改变了 JFH1 株 HCV 亚基因组复制子在 Huh7.5.1 中的复制能力。关于三个众所周知的可转移 HIV-1 病毒元件——Env、Nef 和 Tat-Nef，它们独特地增强了复制子的表达，而 Tat，而不是其他的，显著调节了肝细胞中的候选基因的表达。这些细胞和病毒基因在复制子细胞中的组合表达进一步改变了复制子的表达。总之，这些结果表明，HIV-1 病毒蛋白可以通过不同的分子机制加重合并感染患者的肝病理，直接或间接地使 HCV 复制失调，即使在血友病患者中报道了 HCV 载量与终末期肝病之间缺乏关联，并且调节对疾病进展至关重要的肝细胞基因的表达。这些发现还为 HCV 介导的肝病的改善诊断和预后提供了重要的肝细胞生物标志物的研究进展。
