Bile‐derived organoids from patients with primary sclerosing cholangitis recapitulate their inflammatory immune profile

Abstract

Primary sclerosing cholangitis (PSC) is a heterogeneous and progressive fibroinflammatory cholangiopathy with no known etiology or effective treatment. Studies of PSC are limited due to difficulty in accessing the cholangiocyte, the small percentage of these cells in the liver, instability of in vitro culture systems, and reliance on samples from end‐stage disease. Here we demonstrate that stem cells can be isolated from the bile of PSC patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) earlier in their clinical course, and maintained long‐term in vitro as 3D organoids that express a biliary genetic phenotype. Additionally, bile‐derived organoids (BDOs) can be biobanked and samples obtained longitudinally over the course of the disease. These BDOs express known cholangiocyte markers including gamma glutamyl transferase, cytokeratin 19, epithelial cellular adhesion molecule, cystic fibrosis transmembrane conductance regulator, and anion exchanger 2. RNA sequence analysis identified 39 genes whose expression differed in organoids from PSC patients as compared to non‐PSC controls, including HLA‐DMA and CCL20, immune‐related genes previously described in GWAS studies of PSC. Incubation of these BDOs with IL17A or TNFα lead to an immune reactive phenotype with a significant increase in secretion of proinflammatory mediators, including CCL20, a T‐cell chemoattractant. Conclusions: This study demonstrates that bile can be used as a source of biliary‐like cells that can be maintained long‐term in vitro as 3D organoids. These BDOs retain features of cholangiopathies, including the ability to react to inflammatory stimuli by secreting chemokines, and propagating an immune reactive phenotype reflective of the pathogenesis of these diseases. Thus, BDOs represent a novel platform for the study of the pathogenesis and therapy of cholangiopathies, particularly PSC.

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