Human hepatocyte-enriched miRNA-192-3p promotes HBV replication through inhibiting Akt/mTOR signalling by targeting ZNF143 in hepatic cell lines

Emerg Microbes Infect. 2022 Dec;11(1):616-628. doi: 10.1080/22221751.2022.2037393.

Abstract

Previous studies have revealed multiple tissue- or cell-specific or enriched miRNA profiles. However, miRNA profiles enriched in hepatic cell types and their effect on HBV replication have not been well elucidated. In this study, primary human hepatocytes (PHHs), Kupffer cells (KCs), liver sinusoidal endothelial cells (LSECs), and hepatic stellate cells (HSCs) were prepared from liver specimens of non-HBV-infected patients. Four hepatic cell type-enriched miRNA profiles were identified from purified liver cells miRNA microarray assay. The results revealed that 12 miRNAs, including miR-122-5p and miR-192-3p were PHH-enriched; 9 miRNAs, including miR-142-5p and miR-155-5p were KC-enriched; 6 miRNAs, including miR-126-3p and miR-222-3p were LSEC-enriched; and 14 miRNAs, including miR-214-3p and miR-199a-3p were HSC-enriched. By testing the effect of 11 PHH-enriched miRNAs on HBV production, we observed that miR-192-3p had the greatest pro-virus effect in hepatic cell lines. Moreover, we further found that miR-192-3p promoted HBV replication and gene expression through inhibiting Akt/mTOR signalling by direct targeting of ZNF143 in HepG2.2.15 cells. Additionally, the serum and hepatic miR-192-3p expression levels were significantly higher in chronic hepatitis B patients than in healthy controls and serum miR-192-3p positively correlated with the serum levels of HBV DNA and HBsAg. Collectively, we identified miRNA profiles enriched in four hepatic cell types and revealed that PHH-enriched miR-192-3p promoted HBV replication through inhibiting Akt/mTOR signalling by direct targeting of ZNF143 in hepatic cell lines. Our study provides a specific perspective for the role of hepatic cell type-enriched miRNA in interaction with viral replication and various liver pathogenesis.

Keywords: Abbreviations: CMIA: chemiluminescent microparticle immunoassay; Akt/mTOR signalling; HBV: hepatitis B virus; HSC: hepatic stellate cell; KC: Kupffer cell; LSEC: liver sinusoidal endothelial cell; MiRNA profile; PHH: primary human hepatocyte; RI: replicative intermediate; ZNF143; hepatitis B virus; miR-192-3p; pgRNA: pregenomic RNA.

MeSH terms

  • Endothelial Cells / metabolism
  • Hepatitis B virus* / genetics
  • Hepatitis B virus* / metabolism
  • Hepatocytes / metabolism
  • Humans
  • Liver / pathology
  • MicroRNAs* / metabolism
  • Proto-Oncogene Proteins c-akt / genetics
  • TOR Serine-Threonine Kinases / genetics
  • Trans-Activators

Substances

  • MIRN192 microRNA, human
  • MIRN214 microRNA, human
  • MicroRNAs
  • Trans-Activators
  • ZNF143 protein, human
  • MTOR protein, human
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases

Grants and funding

This work was supported by the National Natural Science Foundation of China (81871640, 82172255, 81800526, 82002131, 81801999), the Shanghai Shen Kang Hospital Development Center (SHDC12019116), the Shanghai Key Clinical Specialty Construction Program (ZK2019B24), and Shang Rao Science and Technology Department (2020D001). F.L. received a scholarship from the Medizinische Fakultat Universitat Duisburg Essen.