Abstract :
[en] Liver transplantation is the best treatment for end-stage liver failure and primary liver cancer, but approximately one third of the recipients will develop ischemic cholangiopathy (IC). This biliary complication raises morbidity, mortality, and healthcare costs. Preservation injury is the main cause of IC. Indeed, 87% of livers exhibits biliary epithelial damage at the end of cold preservation. The limited understanding of cholangiocyte preservation injury is a significant barrier to preventing and treating IC. To address this gap, we will use cholangiocyte organoids, which replicate the architecture and function of biliary tissue, and can be cultured retaining disease-specific characteristics. Our research aims to establish cholangiocyte organoids as preclinical model to investigate the pathophysiology of cholangiocyte injury during liver preservation and transplantation. The specific aims are: 1. Generate and characterize porcine and human cholangiocyte organoids. A comparison between DBD and DCD will be established. Cholangiocytes will be cultured in a matrigel solution and expand organoid lines for 3 passages. Organoids will be characterized by, IF, qRT-PCR, transcriptomic, and by lipidomic to identify cholangiocyte markers expression and function. 2. Unravel the
pathophysiology of cholangiocyte injury. Porcine livers will undergo 30 or 60min of anoxia and subsequent 12h reperfusion, mimicking preservation injury. Gallbladder samples at baseline, after anoxia, and 0, 1, 6, and 12h of reperfusion will be used to generate organoids and to compare proliferation capacity, and transcriptome for a comprehensive fingerprint. The long-term goal is to identify actionable targets in the pathophysiology of
cholangiocyte preservation injury, leading to effectively preventing or treating IC.
