255-OR: Therapeutic Potential of Intestinal Epithelial Organoids–Derived Exosomes in Ameliorating Steatohepatitis

Introduction and Objective: Non-alcoholic steatohepatitis is a prevalent metabolic liver disease with limited therapeutic options. This study aimed to investigate the therapeutic potential of intestinal epithelial organoid (IEO)-derived exosomes in treating steatohepatitis. Methods: IEOs were generated from the small intestinal tissue of eight-week-old C57BL/6 mice, and exosomes were isolated from IEO culture supernatants. Human hepatoma HepG2 cells were treated with IEO-derived exosomes and palmitic acid (PA). Additionally, the role of IEO-derived exosomes in macrophages-mediated inflammation was evaluated using human leukemia THP-1 cells treated with PA and lipopolysaccharide (LPS), a model for macrophage-mediated inflammation in steatohepatitis. Experimental analyses included cell viability assays (CCK-8), lipid accumulation assessments (triglycerides content and Oil Red O staining), and quantitative PCR for fibrosis and inflammatory markers. Results: IEO-derived exosome treatment significantly attenuated PA-treated cell death in HepG2 cells, thereby enhancing cell viability. Oil Red O staining and TG content revealed that IEO-derived exosomes significantly attenuated lipotoxicity-induced lipid accumulation in hepatocytes. mRNA expression of fibrosis markers (COL1A1, ACTA2) and inflammatory markers (NLRP3, TNF-α) was significantly downregulated in PA-treated HepG2 cells following exosome treatment. In PA+LPS-treated THP-1 cells, IEO-derived exosomes also reduced the expression of inflammatory cytokines (TNF-α, IL-6, IL-8, CXCL10, and IL-1β) and fibrosis markers (COL1A1, TGF- β1). Conclusion: These findings demonstrate that IEO-derived exosomes attenuate lipotoxicity-induced steatosis, fibrosis, and macrophage-mediated inflammation, highlighting their therapeutic potential for steatohepatitis through modulation of gut-liver axis crosstalk. Disclosure S. Hahn: None. J. Kim: None. G. Kim: None.