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Updated: Dec 6, 2025
Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery
Published on: May 19, 2018
High-Fidelity Drug-Induced Liver Injury Screen Using Human Pluripotent Stem Cell-Derived Organoids
Tadahiro Shinozawa1, Masaki Kimura1, Yuqi Cai1
1Division of Gastroenterology, Hepatology & Nutrition, Developmental Biology and Center for Stem Cell and Organoid Medicine (CuSTOM), Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
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 multiplexed readouts measuring viability, cholestatic and/or mitochondrial toxicity with high predictive values for 238 marketed drugs at 4 different concentrations (Sensitivity: 88.7%, Specificity: 88.9%). LoT positively predicts genomic predisposition (CYP2C9∗2) for Bosentan-induced cholestasis.
Conclusions:
Liver organoid-based Toxicity screen (LoT) is a potential assay system for liver toxicology studies, facilitating compound optimization, mechanistic study, and precision medicine as well as drug screening applications.
Insights
A new human liver organoid model accurately predicts drug-induced liver injury (DILI) and individual susceptibility. This organoid-based toxicity screen (LoT) aids drug development and precision medicine.
Area of Science:
- Drug discovery and development
- Toxicology
- Regenerative medicine
Background:
- Predicting drug-induced liver injury (DILI) in preclinical stages is a significant challenge.
- A need exists for human systems to study DILI mechanisms and individual drug susceptibility.
Purpose of the Study:
- Establish a human liver organoid (HLO)-based screening model for DILI.
- Analyze DILI pathology at the organoid level.
- Develop a predictive model for drug toxicology.
Main Methods:
- Generated reproducible HLOs from pluripotent stem cell lines with bile transport function.
- Utilized qRT-PCR and single-cell RNA-seq to analyze hepatocyte transcriptomic state.
- Developed a high-throughput imaging platform for HLO-based DILI assays.
Main Results:
- HLOs exhibited polarized hepatocytes and bile canaliculi-like structures with unidirectional bile acid transport.
- Single-cell RNA-seq revealed diverse hepatocytic populations mirroring primary hepatocytes.
- The developed organoid-based toxicity screen (LoT) achieved high predictive values (Sensitivity: 88.7%, Specificity: 88.9%) for 238 drugs.
Conclusions:
- The liver organoid-based Toxicity screen (LoT) is a promising system for liver toxicology.
- LoT facilitates compound optimization, mechanistic studies, and drug screening.
- This model supports precision medicine by predicting individual drug responses.
