Modeling bile duct ischemia and reoxygenation injury inhuman cholangiocyte organoids for screening of novel cholangio-protective agents.

Abstract

Ischemia of the bile duct is a common feature in liver disease and transplantation, which represents a major cause of morbidity and mortality, especially after liver transplantation. Detailed knowledge of its pathogenesis remains incomplete due to the lack of appropriate invitro models. To recapitulate biliary damage induced by ischemia and reperfusion invitro, human intrahepatic cholangiocyte organoids (ICOs) were grown at low oxygen levels of 1% up to 72h, followed by re-oxygenation at normal levels. ICOs stressed by ischemia and subsequent re-oxygenation represented the dynamic change in biliary cell proliferation, upregulation of epithelial-mesenchymal transition (EMT)-associated markers, and the evocation of phase-dependent cell death programs similar to what is described in patients. Clinical-grade alpha-1 antitrypsin was identified as a potent inhibitor of both ischemia-induced apoptosis and necroptosis. These findings demonstrate that ICOs recapitulate ischemic cholangiopathy invitro and enable drug assessment studies for the discovery of new therapeutics for ischemic cholangiopathies. Dutch Digestive FoundationMLDS D16-26; TKI-LSH (Topconsortium Kennis en Innovatie-Life Sciences & Health) grant RELOAD, EMC-LSH19002; Medical Delta program "Regenerative Medicine 4D"; China Scholarship Council No. 201706230252.