A cell culture system for distinguishing hepatitis C viruses with and without liver cancer-related mutations in the viral core gene

Ahmed El-Shamy, Francis J. Eng, Erin H. Doyle, Arielle L. Klepper, Xiaochen Sun, Angelo Sangiovanni, Massimo Iavarone, Massimo Colombo, Robert E. Schwartz, Yujin Hoshida, Andrea D. Branch

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Background & Aims 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. Methods HuH7 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). Results 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. Conclusions 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.

Original languageEnglish
Pages (from-to)1323-1333
Number of pages11
JournalJournal of Hepatology
Volume63
Issue number6
DOIs
Publication statusPublished - Dec 1 2015

Fingerprint

Viral Genes
Liver Neoplasms
Hepacivirus
Cell Culture Techniques
Mutation
Viruses
Hepatocytes
Hepatocellular Carcinoma
Genes
Gene Expression
Oxidative Phosphorylation
Serum
Transcriptome
Type 2 Diabetes Mellitus
Insulin Resistance
Fibrosis
Genotype
Carcinoma
Biopsy
Liver

Keywords

  • Core-mutations
  • HCC
  • HCV
  • Human serum

ASJC Scopus subject areas

  • Medicine(all)
  • Hepatology

Cite this

A cell culture system for distinguishing hepatitis C viruses with and without liver cancer-related mutations in the viral core gene. / El-Shamy, Ahmed; Eng, Francis J.; Doyle, Erin H.; Klepper, Arielle L.; Sun, Xiaochen; Sangiovanni, Angelo; Iavarone, Massimo; Colombo, Massimo; Schwartz, Robert E.; Hoshida, Yujin; Branch, Andrea D.

In: Journal of Hepatology, Vol. 63, No. 6, 01.12.2015, p. 1323-1333.

Research output: Contribution to journalArticle

El-Shamy, Ahmed ; Eng, Francis J. ; Doyle, Erin H. ; Klepper, Arielle L. ; Sun, Xiaochen ; Sangiovanni, Angelo ; Iavarone, Massimo ; Colombo, Massimo ; Schwartz, Robert E. ; Hoshida, Yujin ; Branch, Andrea D. / A cell culture system for distinguishing hepatitis C viruses with and without liver cancer-related mutations in the viral core gene. In: Journal of Hepatology. 2015 ; Vol. 63, No. 6. pp. 1323-1333.
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AU - El-Shamy, Ahmed

AU - Eng, Francis J.

AU - Doyle, Erin H.

AU - Klepper, Arielle L.

AU - Sun, Xiaochen

AU - Sangiovanni, Angelo

AU - Iavarone, Massimo

AU - Colombo, Massimo

AU - Schwartz, Robert E.

AU - Hoshida, Yujin

AU - Branch, Andrea D.

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N2 - Background & Aims 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. Methods HuH7 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). Results 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. Conclusions 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.

AB - Background & Aims 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. Methods HuH7 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). Results 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. Conclusions 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.

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KW - HCC

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