Adipose Tissue Macrophages in Metabolic Dysfunction-Associated Steatohepatitis Secrete Extracellular Vesicles That Activate Liver Fibrosis in Obese Male Mice

Gastroenterology. 2025 Sep;169(4):691-704.e9. doi: 10.1053/j.gastro.2025.03.033. Epub 2025 Apr 9.

Abstract

Background & aims: Given the need for effective interventions in metabolic dysfunction-associated steatohepatitis (MASH), understanding the role of adipose tissue macrophage (ATM)-derived small extracellular vesicles (sEVs) is important. We aimed to evaluate the contribution of MASH-ATM-sEVs to the development of liver fibrosis in obese male mice.

Methods: Using flow cytometry and nanoparticle tracking analysis, we characterized MASH-ATMs and their secreted sEVs. We assessed the fibrogenic effects of sEVs from MASH-ATMs or anti-inflammatory macrophages on stellate cells in vitro and in mice in vivo. In addition, we isolated Dicer knockdown microRNA (miRNA)-depleted sEVs from MASH-ATMs and cotreated stellate cells with MASH-ATM-sEVs and miR-155 or miR-34a antagomirs.

Results: MASH-ATMs exhibited a pro-inflammatory and lipid-associated phenotype, secreting sEVs enriched in the fibrogenic miRNAs, miR-155 and miR-34a, which also down-regulate Pparg. In vitro, MASH-ATM-sEVs induced hepatic stellate cell activation and fibrogenesis and exacerbated liver fibrosis when administered to obese mice. In addition, anti-inflammatory macrophage sEVs mitigated fibrosis both in vitro and in vivo. miRNA-free Dicer knockdown-MASH-ATM-sEVs were without effects and cotreatment with miR-155/miR-34a antagomirs blocked the effects of MASH-ATM-sEVs to induce hepatic stellate cell activation.

Conclusions: This study demonstrated the role of MASH-ATM-sEVs in promoting liver fibrosis in obesity. Identification of the fibrogenic miRs, miR-155, and miR-34a, within MASH-ATM-sEVs, highlights the mechanistic importance of extrahepatic signals in MASH. These findings showed the therapeutic potential of modulating macrophage phenotypes and their sEV cargo to ameliorate MASH.

Keywords: Adipose Tissue Macrophages; Exosomes; Lipid-Associated Macrophages; MASH; MicroRNA.

MeSH terms

  • Adipose Tissue* / cytology
  • Adipose Tissue* / metabolism
  • Adipose Tissue* / pathology
  • Animals
  • Cells, Cultured
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism
  • Disease Models, Animal
  • Extracellular Vesicles* / metabolism
  • Extracellular Vesicles* / pathology
  • Hepatic Stellate Cells / metabolism
  • Hepatic Stellate Cells / pathology
  • Liver Cirrhosis* / etiology
  • Liver Cirrhosis* / genetics
  • Liver Cirrhosis* / metabolism
  • Liver Cirrhosis* / pathology
  • Liver* / metabolism
  • Liver* / pathology
  • Macrophages* / metabolism
  • Macrophages* / pathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / genetics
  • MicroRNAs / metabolism
  • Non-alcoholic Fatty Liver Disease* / genetics
  • Non-alcoholic Fatty Liver Disease* / metabolism
  • Non-alcoholic Fatty Liver Disease* / pathology
  • Obesity* / complications
  • Obesity* / metabolism
  • Obesity* / pathology
  • Ribonuclease III / genetics
  • Ribonuclease III / metabolism

Substances

  • MicroRNAs
  • MIRN34a microRNA, mouse
  • Ribonuclease III
  • Mirn155 microRNA, mouse
  • Dicer1 protein, mouse
  • DEAD-box RNA Helicases