miR-122 inhibition in a human liver organoid model leads to liver inflammation, necrosis, steatofibrosis and dysregulated insulin signaling

PLoS One. 2018 Jul 19;13(7):e0200847. doi: 10.1371/journal.pone.0200847. eCollection 2018.


To investigate the role of miR-122 in the development and regression of non-alcoholic fatty liver disease (NAFLD) in vitro, we used multicellular 3D human liver organoids developed in our laboratory. These organoids consist of primary human hepatocytes, Kupffer cells, quiescent stellate cells and liver sinusoidal endothelial cells. They remain viable and functional for 4 weeks expressing typical markers of liver function such as synthesis of albumin, urea, and alpha-1 p450 drug metabolism. Before mixing, hepatic cells were transduced with lentivirus to inhibit miR122 expression (ABM, CA). Immediately after the organoids were fully formed (day 4) or after 1 or 2 weeks of additional incubation (days 11 or 18), the organoids were analyzed using fluorescent live/dead staining and ATP production; total RNA was extracted for qPCR gene expression profiling. Our results show that miR-122 inhibition in liver organoids leads to inflammation, necrosis, steatosis and fibrosis. This was associated with increase in inflammatory cytokines (IL6, TNF), chemokines (CCL2, CCL3) and increase in a subset of Matrix Metaloproteinases (MMP8, MMP9). An altered expression of key genes in lipid metabolism (i.e LPL, LDLR) and insulin signaling (i.e GLUT4, IRS1) was also identified.

Conclusion: Our results highlight the role of miR-122 inhibition in liver inflammation, steatofibrosis and dysregulation of insulin signaling. Patients with NAFLD are known to have altered levels of miR-122, therefore we suggest that miR-122 mimics could play a useful role in reversing liver steatofibrosis and insulin resistance seen in patients with NAFLD.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Chemokine CCL2 / metabolism
  • Chemokine CCL3 / metabolism
  • Glucose Transporter Type 4 / metabolism
  • Hepatocytes / metabolism
  • Humans
  • Inflammation / metabolism*
  • Insulin / metabolism*
  • Insulin Receptor Substrate Proteins / metabolism
  • Interleukin-6 / metabolism
  • Kupffer Cells / metabolism
  • Liver / cytology*
  • Liver / metabolism*
  • Matrix Metalloproteinase 8 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • MicroRNAs / metabolism*
  • Necrosis / metabolism*
  • Non-alcoholic Fatty Liver Disease / metabolism*
  • Organoids / cytology*
  • Organoids / metabolism*
  • Signal Transduction


  • Chemokine CCL2
  • Chemokine CCL3
  • Glucose Transporter Type 4
  • IRS1 protein, human
  • Insulin
  • Insulin Receptor Substrate Proteins
  • Interleukin-6
  • MIRN122 microRNA, human
  • MicroRNAs
  • Matrix Metalloproteinase 8
  • Matrix Metalloproteinase 9

Grants and funding

We greatly thank Defense Threat Reduction Agency Contracts N66001-13-C-2027 (AA & CEB) and NIH R01DK110167-01A1 (CEB) R15 HL 117199 (HLB), U54 DK 083909 9 (HLB). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.