Patient-derived liver organoids recapitulate epithelial heterogeneity and enable precision disease modelling of alcohol-associated liver disease
Organoids are emerging as a powerful human-based in vitro tool in the biomedical field. However, patient-derived liver organoids fail to recapitulate the liver epithelial heterogeneity and its generation still requires liver  surgical resections, thus limiting personalized chronic liver diseases modeling. Here, we report the derivation of organoids from intact liver needle biopsies(b-Orgs) from alcohol-associated liver disease (ALD) patients for precision disease modeling and drug testing. B-Orgs were generated with an efficiency of 80% from patients with early and advanced stages of ALD. b-Orgs show an enriched hepatocyte phenotype as assessed by immunofluorescence, functional studies, and transcriptomics. Single cell RNA-sequencing revealed a heterogeneous epithelial composition comprising hepatocyte, biliary and progenitor hepatobiliary populations, mirroring the epithelial populations found in advanced ALD patients. Moreover, b-Orgs preserve disease-stage features, as b-Orgs from advanced ALD patients showed increased expression of genes related to epithelial-mesenchymal transition, angiogenesis and inflammation. Stimulation of b-Orgs with ethanol and pro-inflammatory mediators,promoted ALD features such asROS production, lipid accumulation, inflammation and decreased proliferation, which were mitigated in response to prednisolone. Overall, we provide a new methodology to obtain b-Orgs showing epithelial complexity and patient specific features, thus expanding organoid-based liver disease modelling for personalized medicine. Overall design: single-cell RNA sequencing 3' (scRNA-seq)