Hepatocyte-derived extracellular vesicles promote endothelial dedifferentiation in chronic liver disease through the miR-153-3p-pyroptosis axis.

BACKGROUND AND AIMS: Liver cells communicate paracrinally through extracellular vesicles (EVs). This study investigated the role of microRNAs (miRNAs) embedded in hepatocyte-derived EVs (hepEVs) modulating endothelial cell dedifferentiation in chronic liver disease.

APPROACH AND

RESULTS

EVs were purified from primary hepatocytes isolated from healthy (CT) and cirrhotic (CH) livers in both humans and rats. CT-rats were intravenously treated with fluorescence-labeled hepEVs-CT or hepEVs-CH (200 μg/d, 3 d, n=10/group) to assess biodistribution and LSECs phenotype. miRNA profiling of human hepEVs-CH was validated in rat models, and commonly altered miRNAs were overexpressed in CT-LSECs. Gene dysregulation was analyzed in primary CH-LSECs and liver tissues from rats and humans with cirrhosis, as well as CT-LSECs treated with miRNA-specific upregulated hepEVs. CH-rats were treated with the Caspase-1 inhibitor VX-765 (15 mg/kg/d, 10 doses, n=11/group) to evaluate its effects on liver function, hemodynamics, and endothelial phenotype. In vivo, hepEVs-CH accumulated predominantly in LSECs, disrupting genes involved in fibrosis, inflammation, and cell death, while enhancing the von Willebrand factor expression and altering eNOS activity. Profiling of human hepEVs-CH revealed 37 dysregulated miRNAs, with miR-200a-3p and miR-153-3p validated in rat hepEVs-CH. Transfection of miR-153-3p into CT-LSEC dysregulated 771 genes involved in inflammation and cell death, particularly pyroptosis. Increased active Caspase-1 and Gasdermin-D expression was observed in human and rat cirrhotic liver tissue, as well as in CT-LSEC treated with miR-153-3p-hepEVs, indicating enhanced pyroptosis. In vivo, VX-765 treatment reduced hepatic pyroptosis, improving endothelial phenotype and portal hypertension.

CONCLUSIONS

HepEVs-derived miRNAs, particularly miR-153-3p, contribute to endothelial dysfunction in chronic liver disease by triggering pyroptosis through a paracrine mechanism. Inhibiting Caspase-1 may provide a novel therapeutic approach to mitigate endothelial dysfunction in chronic liver disease.