Farnesoid X Receptor Agonist INT ‐787 Protects Human Liver Organoids From Alcohol‐Induced Injury

ABSTRACT Background Alcohol‐associated liver disease (ALD) represents a spectrum of liver injury ranging from steatosis to alcohol‐associated hepatitis, cirrhosis, and hepatocellular carcinoma. Activation of the farnesoid X receptor (FXR) decreases liver steatosis, inflammation, and fibrosis in animal models of liver‐related diseases, including cirrhosis and alcohol‐induced liver injury, and in patients with primary biliary cholangitis or metabolic dysfunction‐associated steatohepatitis. We assessed the therapeutic potential of the bile acid analogue and potent FXR agonist INT‐787 to determine its effects on alcohol‐induced liver injury using an in vitro human liver organoid model. Methods Human liver organoids were obtained from multidonor primary human hepatocytes cocultured with primary Kupffer cells and liver endothelial cells. Organoids were cultured with alcohol 0.5% in the presence of dimethyl sulfoxide (control), INT‐787, or the DNA methyltransferase inhibitor larsucosterol (DUR‐928). On day 4, steatosis (Nile Red staining) and markers of inflammation (IL‐6 and TNF‐α) and fibrosis (TGF‐β1, αSMA, COL1A1, COL3A1, CTGF and THBS1) were assessed, as well as cell viability (ATP), response to oxidative stress (CYP2E1 and COX‐2), apoptotic pathways (BAX and BCL2), cholestasis (MRP2), and primary bile acid levels. Results INT‐787 counteracts the toxic effect of alcohol exposure by markedly reducing lipid accumulation and significantly decreasing markers of inflammation, fibrosis, oxidative stress, apoptosis, and cholestasis. INT‐787 and DUR‐928 similarly inhibit inflammation, oxidative stress, and apoptosis induced by alcohol while ameliorating cell viability, but INT‐787 is significantly superior to DUR‐928 in inhibiting steatosis and fibrosis. Conclusion INT‐787 represents a promising molecule for the clinical treatment of ALD.