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Updated: Jan 3, 2026
Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery
Published on: May 19, 2018
High-Throughput Screening to Evaluate Inhibition of Bile Acid Transporters Using Human Hepatocytes Isolated From
Hiroshi Kohara1, Piyush Bajaj2, Kazunori Yamanaka1
1Drug Safety Research and Evaluation, Takeda Pharmaceutical Company Limited, Kanagawa 251-8555, Kanagawa, Japan.
Abstract:
Cholestasis resulting from hepatic bile acid efflux transporter inhibition may contribute to drug-induced liver injury (DILI). This condition is a common safety-related reason for drug attrition and withdrawal. To screen for safety risks associated with efflux transport inhibition, we developed a high-throughput cellular assay for different drug discovery phases. Hepatocytes isolated from chimeric mice with humanized livers presented gene expression resembling that of the human liver and demonstrated apical membrane polarity when sandwiched between Matrigel and collagen. The fluorescent bile acid-derivative cholyl-l-lysyl-fluorescein (CLF) was used to quantify drug-induced efflux transport inhibition in hepatocytes. Cyclosporine inhibited CLF accumulation in the apical bile canalicular lumen in a concentration-dependent manner. The assay had equivalent predictive power to a primary human hepatocyte-based assay and greater predictive power than an assay performed with rat hepatocytes. Predictive power was tested using 45 pharmaceutical compounds, and 91.3% of the compounds with cholestatic potential (21/23) had margins (IC50/Cmax) < 20. In contrast, 90.9% (20/22) of compounds without cholestatic potential had IC50/Cmax>20. Assay sensitivity and specificity were 91.3% and 90.9%, respectively. We suggest that this improved assay performance could result from higher expression of efflux transporters, metabolic pathways, and/or species differences. Given the long-term supply of cells from the same donor, the humanized mouse-derived hepatocyte-based CLF efflux assay could be a valuable tool for predicting cholestatic DILI.
Insights
A new assay using humanized mouse liver cells accurately predicts drug-induced liver injury (DILI) caused by bile acid transport issues. This tool aids early drug safety screening, improving prediction of cholestatic DILI risks.
Area of Science:
- Hepatology and Drug Metabolism
- Toxicology and Safety Pharmacology
Background:
- Drug-induced liver injury (DILI) is a major cause of drug attrition, often linked to impaired bile acid transport.
- Developing reliable assays to predict cholestatic DILI is crucial for drug discovery.
- Current assays using rat hepatocytes or primary human hepatocytes have limitations.
Purpose of the Study:
- To develop and validate a high-throughput cellular assay for screening drug-induced efflux transport inhibition.
- To assess the predictive power of this novel assay for cholestatic DILI.
Main Methods:
- Utilized hepatocytes from chimeric mice with humanized livers, cultured in a sandwich model.
- Employed cholyl-l-lysyl-fluorescein (CLF), a fluorescent bile acid analog, to measure efflux transport.
- Validated the assay with 45 pharmaceutical compounds, comparing IC50/Cmax margins to known cholestatic potential.
Main Results:
- The assay demonstrated high predictive power, with 91.3% accuracy for compounds with cholestatic potential and 90.9% for those without.
- Sensitivity and specificity were 91.3% and 90.9%, respectively.
- The humanized mouse hepatocyte assay showed superior predictive capability compared to rat hepatocyte assays.
Conclusions:
- A humanized mouse-derived hepatocyte CLF efflux assay is a valuable tool for predicting cholestatic DILI.
- The assay's performance may stem from higher expression of efflux transporters and metabolic pathways.
- This assay offers a reliable method for early-stage drug safety assessment, reducing DILI risks.
