High-Fidelity Drug-Induced Liver Injury Screen Using Human Pluripotent Stem Cell–Derived Organoids

作者：Tadahiro Shinozawa, Masaki Kimura, Yuqi Cai, Norikazu Saiki, Yosuke Yoneyama, Rie Ouchi, Hiroyuki Koike, Mari Maezawa, Ranran Zhang, Andrew Dunn, Autumn Ferguson, Shodai Togo, Kyle Lewis, Wendy L. Thompson, Akihiro Asai, Takanori Takebe · 发表于：Gastroenterology · 年份：2020 · DOI：10.1053/j.gastro.2020.10.002 · 被引用次数：364 · 研究领域：Liver physiology and pathology、Pluripotent Stem Cells Research、Drug-Induced Hepatotoxicity and Protection

BACKGROUND & AIMS: Preclinical identification of compounds at risk of causing drug induced liver injury (DILI) remains a significant challenge in drug development, highlighting a need for a predictive human system to study complicated DILI mechanism and susceptibility to individual drug. Here, we established a human liver organoid (HLO)-based screening model for analyzing DILI pathology at organoid resolution. METHODS: We first developed a reproducible method to generate HLO from storable foregut progenitors from pluripotent stem cell (PSC) lines with reproducible bile transport function. The qRT-PCR and single cell RNA-seq determined hepatocyte transcriptomic state in cells of HLO relative to primary hepatocytes. Histological and ultrastructural analyses were performed to evaluate micro-anatomical architecture. HLO based drug-induced liver injury assays were transformed into a 384 well based high-speed live imaging platform. RESULTS: HLO, generated from 10 different pluripotent stem cell lines, contain polarized immature hepatocytes with bile canaliculi-like architecture, establishing the unidirectional bile acid transport pathway. Single cell RNA-seq profiling identified diverse and zonal hepatocytic populations that in part emulate primary adult hepatocytes. The accumulation of fluorescent bile acid into organoid was impaired by CRISPR-Cas9-based gene editing and transporter inhibitor treatment with BSEP. Furthermore, we successfully developed an organoid based assay with ...