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Combined analysis of metabolomes, proteomes, and transcriptomes of HCV-infected cells and liver to identify pathways associated with disease development

Authors

  • J. Lupberger
  • T. Croonenborghs
  • A.A. Roca Suarez
  • N. Van Renne
  • F. Jühling
  • M.A. Oudot
  • A. Virzì
  • S. Bandiera
  • C. Jamey
  • G. Meszaros
  • D. Brumaru
  • A. Mukherji
  • S.C. Durand
  • L. Heydmann
  • E.R. Verrier
  • H. El Saghire
  • N. Hamdane
  • R. Bartenschlager
  • S. Fereshetian
  • E. Ramberger
  • R. Sinha
  • M. Nabian
  • C. Everaert
  • M. Jovanovic
  • P. Mertins
  • S.A. Carr
  • K. Chayama
  • N. Dali-Youcef
  • R. Ricci
  • N.M. Bardeesy
  • N. Fujiwara
  • O. Gevaert
  • M.B. Zeisel
  • Y. Hoshida
  • N. Pochet
  • T.F. Baumert

Journal

  • Gastroenterology

Citation

  • Gastroenterology

Abstract

  • BACKGROUND & AIMS: The mechanisms of hepatitis C virus (HCV) infection, liver disease progression, and hepatocarcinogenesis are only partially understood. We performed genomic, proteomic, and metabolomic analyses of HCV-infected cells and chimeric mice to learn more about these processes. METHODS: Huh7.5.1(dif) (hepatocyte-like cells) were infected with culture-derived HCV and used in RNA-Seq, proteomic, metabolomic, and integrative genomic analyses. uPA/SCID mice were injected with serum from HCV-infected patients; 8 weeks later, liver tissues were collected and analyzed by RNA-seq and proteomics. Using differential expression, gene set enrichment analyses, and protein interaction mapping, we identified pathways that changed in response to HCV infection. We validated our findings in studies of liver tissues from 216 patients with HCV infection and early-stage cirrhosis and paired biopsies from 99 patients with hepatocellular carcinoma, including 17 patients with histologic features of steatohepatitis. Cirrhotic liver tissues from patients with HCV infection were classified into 2 groups based on relative peroxisome function; outcomes assessed included Child-Pugh class, development of hepatocellular carcinoma, survival and steatohepatitis. Hepatocellular carcinomas were classified according to steatohepatitis; the outcome was relative peroxisomal function. RESULTS: We quantified 21,950 mRNAs and 8297 proteins in HCV-infected cells. Upon HCV infection of hepatocyte-like cells and chimeric mice, we observed significant changes in levels of mRNAs and proteins involved in metabolism and hepatocarcinogenesis. HCV infection of hepatocyte-like cells significantly increased levels of mRNAs, but not proteins, that regulate the innate immune response-we believe this was due to the inhibition of translation in these cells. HCV infection of hepatocyte-like cells increased glucose consumption and metabolism and the STAT3 signaling pathway and reduced peroxisome function. Peroxisomes mediate beta-oxidation of very long-chain fatty acids (VLCFAs); we found intracellular accumulation of VLCFAs in HCV-infected cells, which is also observed in patients with fatty liver disease. Cells in livers from HCV-infected mice had significant reductions in levels of mRNAs and proteins associated with peroxisome function, indication perturbation of peroxisomes. We associated defects in peroxisome function with outcomes and features of HCV-associated cirrhosis, fatty liver disease, and hepatocellular carcinoma in patients. CONCLUSIONS: We performed combined transcriptome, proteome, and metabolome analyses of liver tissues from HCV-infected hepatocyte-like cells and HCV-infected mice. We found that HCV infection increases glucose metabolism and the STAT3 signaling pathway and thereby reduces peroxisome function; alterations in expression of peroxisome genes were associated with outcomes of patients with liver diseases. These findings provide insights into liver disease pathogenesis and might be used to identify new therapeutic targets.


DOI

doi:10.1053/j.gastro.2019.04.003