Functional microRNA screen uncovers O-linked N-acetylglucosamine transferase as a host factor modulating hepatitis C virus morphogenesis and infectivity

Gut. 2020 Feb;69(2):380-392. doi: 10.1136/gutjnl-2018-317423. Epub 2019 May 10.

Abstract

Objective: Infection of human hepatocytes by the hepatitis C virus (HCV) is a multistep process involving both viral and host factors. microRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate gene expression. Given that miRNAs were indicated to regulate between 30% and 75% of all human genes, we aimed to investigate the functional and regulatory role of miRNAs for the HCV life cycle.

Design: To systematically reveal human miRNAs affecting the HCV life cycle, we performed a two-step functional high-throughput miRNA mimic screen in Huh7.5.1 cells infected with recombinant cell culture-derived HCV. miRNA targeting was then assessed using a combination of computational and functional approaches.

Results: We uncovered miR-501-3p and miR-619-3p as novel modulators of HCV assembly/release. We discovered that these miRNAs regulate O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) protein expression and identified OGT and O-GlcNAcylation as regulators of HCV morphogenesis and infectivity. Furthermore, increased OGT expression in patient-derived liver tissue was associated with HCV-induced liver disease and cancer.

Conclusion: miR-501-3p and miR-619-3p and their target OGT are previously undiscovered regulatory host factors for HCV assembly and infectivity. In addition to its effect on HCV morphogenesis, OGT may play a role in HCV-induced liver disease and hepatocarcinogenesis.

Keywords: HCV; hepatitis C; hepatocyte; molecular mechanisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Regulation / physiology
  • Gene Knockdown Techniques / methods
  • Genome-Wide Association Study / methods
  • Hepacivirus / pathogenicity*
  • Hepacivirus / physiology
  • Hepatitis C, Chronic / genetics*
  • Hepatitis C, Chronic / virology
  • Hepatocytes / virology
  • Host-Pathogen Interactions / genetics
  • Humans
  • Life Cycle Stages / genetics
  • MicroRNAs / genetics
  • Morphogenesis / physiology
  • N-Acetylglucosaminyltransferases / genetics
  • N-Acetylglucosaminyltransferases / physiology*
  • Up-Regulation
  • Virulence / genetics

Substances

  • MIRN-619 microRNA, human
  • MIRN501 microRNA, human
  • MicroRNAs
  • N-Acetylglucosaminyltransferases
  • OGT protein, human