ABSTRACT
The treatment of common bile duct (CBD) disorders, such as biliary atresia or ischemic strictures, is restricted by the lack of biliary tissue from healthy donors suitable for surgical reconstruction. Here we report a new method for the isolation and propagation of human cholangiocytes from the extrahepatic biliary tree in the form of extrahepatic cholangiocyte organoids (ECOs) for regenerative medicine applications. The resulting ECOs closely resemble primary cholangiocytes in terms of their transcriptomic profile and functional properties. We explore the regenerative potential of these organoids in vivo and demonstrate that ECOs self-organize into bile duct-like tubes expressing biliary markers following transplantation under the kidney capsule of immunocompromised mice. In addition, when seeded on biodegradable scaffolds, ECOs form tissue-like structures retaining biliary characteristics. The resulting bioengineered tissue can reconstruct the gallbladder wall and repair the biliary epithelium following transplantation into a mouse model of injury. Furthermore, bioengineered artificial ducts can replace the native CBD, with no evidence of cholestasis or occlusion of the lumen. In conclusion, ECOs can successfully reconstruct the biliary tree, providing proof of principle for organ regeneration using human primary cholangiocytes expanded in vitro.
Subject(s)
Bile Ducts, Extrahepatic/physiology , Epithelial Cells/cytology , Gallbladder/physiology , Organoids/physiology , Regeneration/physiology , Tissue Engineering/methods , Animals , Bile Ducts, Extrahepatic/cytology , Bile Ducts, Extrahepatic/injuries , Biliary Tract/cytology , Biliary Tract/injuries , Biliary Tract/physiology , Cell Transplantation , Cystic Fibrosis Transmembrane Conductance Regulator/metabolism , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Gallbladder/injuries , Humans , In Vitro Techniques , Keratin-19/metabolism , Keratin-7/metabolism , Mice , Organoids/cytology , Organoids/drug effects , Organoids/metabolism , Secretin/pharmacology , Somatostatin/pharmacology , Tissue Scaffolds , gamma-Glutamyltransferase/metabolismABSTRACT
Linking non-coding genetic variants associated with the risk of diseases or disease-relevant traits to target genes is a crucial step to realize GWAS potential in the introduction of precision medicine. Here we set out to determine the mechanisms underpinning variant association with platelet quantitative traits using cell type-matched epigenomic data and promoter long-range interactions. We identify potential regulatory functions for 423 of 565 (75%) non-coding variants associated with platelet traits and we demonstrate, through ex vivo and proof of principle genome editing validation, that variants in super enhancers play an important role in controlling archetypical platelet functions.
