High-Throughput Screening to Evaluate Inhibition of Bile Acid Transporters Using Human Hepatocytes Isolated From Chimeric Mice
| Autor: | Kosuke Harada, Hiroshi Kohara, Yoshiko Okai, Matthew Wagoner, Akimitsu Miyawaki, Toshikatsu Matsui, Kazumasa Miyamoto, Kazunori Yamanaka, Piyush Bajaj, Tadahiro Shinozawa |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
medicine.drug_class Gene Expression Mice Transgenic Toxicology Bile Acids and Salts 03 medical and health sciences Mice 0302 clinical medicine Cholestasis medicine Animals Humans IC50 ATP Binding Cassette Transporter Subfamily B Member 11 biology Bile acid Membrane transport protein Chemistry Cellular Assay Bile Canaliculi Apical membrane medicine.disease Molecular biology High-Throughput Screening Assays 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Hepatocyte biology.protein Cyclosporine Hepatocytes Efflux Chemical and Drug Induced Liver Injury |
| Zdroj: | Toxicological sciences : an official journal of the Society of Toxicology. 173(2) |
| ISSN: | 1096-0929 |
| Popis: | 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. 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. |
| Databáze: | OpenAIRE |
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