Signature molecules expressed differentially in a liver disease stage-specific manner by HIV-1 and HCV co-infection

Abstract

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.

Clinical Morphology of the Liver: Hepatocytes, Endomembrane System

“…На эти же механизмы указывают другие авторы, подтверждающие, что белки ВИЧ-1 могут усугублять поражение печени при ко-инфекции ВГС+ВИЧ путем усиления репликации HCV, модулируя экспрессию гепатоцеллюлярных генов, критических для прогрессирования болезни. Показано, что инактивированный ВИЧ или gp120 увеличивает репликацию HCV, усиливая экспрессию TGF-β1 как в репликоне, так и в инфекционной модели HCV [26,27].…”

Development of a reverse transcription droplet digital PCR (RT-ddPCR) assay for sensitive detection of simian immunodeficiency virus (SIV)

“…One of the most prevalent applications of qRT-PCR is for the detection of viral loads [1][2][3], in which qRT-PCR-enabled data on various causative infectious agents [4] have helped to study disease processes and delineate connections between unique viral sequences and clinical signs and symptoms [5]. Various qRT-PCR assays have been developed to detect and quantify negative-strand RNA viruses (including the measles and mumps viruses, and various viruses that target the respiratory tract) [6,7], positive-strand RNA viruses (including rhino-, entero-and coronaviruses such as SARS-CoV-2) [8][9][10], double-stranded RNA viruses (such as human rotaviruses) [11], and retroviruses (HIV, HTLV, and related viruses in animal models, such as simian immunodeficiency virus (SIV)) [12][13][14][15][16]. In most cases, qRT-PCR assays are more sensitive than traditional methods such as viral culture [17], and have led to a significant increase in the accuracy and clinical relevance of patient testing.…”