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.

A cell culture system for distinguishing hepatitis C viruses with and without liver cancer-related mutations in the viral core gene / A. El Shamy, F.J. Eng, E.H. Doyle, A.L. Klepper, X. Sun, A. Sangiovanni, M. Iavarone, M. Colombo, R.E. Schwartz, Y. Hoshida, A.D. Branch. - In: JOURNAL OF HEPATOLOGY. - ISSN 0168-8278. - 63:6(2015 Dec), pp. 1323-1333. [10.1016/j.jhep.2015.07.024]

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

M. Iavarone;M. Colombo;
2015

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.
Core-mutations; HCC; HCV; Human serum; Carcinoma, Hepatocellular; Cell Culture Techniques; Cell Differentiation; Cell Line; Genetic Variation; Genotype; Hepacivirus; Hepatitis B Core Antigens; Hepatitis C, Chronic; Hepatocytes; Humans; Liver Cirrhosis; Liver Neoplasms; Oncogenes; Prospective Studies; Risk Factors; Transcriptome; Viral Core Proteins; Virulence; Virus Replication; Genes, Viral; Mutation; Medicine (all); Hepatology
Settore MED/12 - Gastroenterologia
dic-2015
Article (author)
File in questo prodotto:
File Dimensione Formato  
1-s2.0-S0168827815005152-main.pdf

accesso riservato

Tipologia: Publisher's version/PDF
Dimensione 1.73 MB
Formato Adobe PDF
1.73 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/437939
Citazioni
  • ???jsp.display-item.citation.pmc??? 14
  • Scopus 20
  • ???jsp.display-item.citation.isi??? 20
social impact