Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, 565-0871, Japan.
Laboratory of Biochemistry and Molecular Biology, School of Pharmaceutical Sciences, Osaka University, Osaka, 565-0871, Japan.
Drug Metab Pharmacokinet. 2020 Jun;35(3):288-296. doi: 10.1016/j.dmpk.2020.01.009. Epub 2020 Feb 10.
Multiple drug resistance 1 (MDR1) is highly expressed in various organs, including the liver, small intestine, and blood-brain barrier (BBB). Because MDR1 plays important roles in the excretion of many drugs, it is necessary to evaluate whether drug candidates are potential substrates of MDR1. Recently, many researchers have shown that human induced pluripotent stem (iPS) cell-derived differentiated cells such as hepatocytes and enterocytes can be applied for pharmacokinetic testing. Here, we attempted to generate MDR1-knockout (KO) iPS cell lines using genome editing technology. The correctly targeted human iPS cell lines were successfully obtained. The expression levels of pluripotent markers in human iPS cells were not changed by MDR1 knockout. The gene expression levels of hepatic markers in MDR1-KO iPS-derived hepatocyte-like cells were higher than those in undifferentiated MDR1-KO iPS cells, suggesting that MDR1-KO iPS cells have hepatic differentiation capacity. In addition, MDR1 expression levels were hardly detected in MDR1-KO iPS cell-derived hepatocyte-like cells. We thus succeeded in establishing MDR1-KO iPS cell lines that could be utilized for pharmacokinetic testing.
多药耐药蛋白 1（MDR1）在包括肝脏、小肠和血脑屏障（BBB）在内的各种器官中高度表达。由于 MDR1 在许多药物的排泄中发挥着重要作用，因此有必要评估候选药物是否为 MDR1 的潜在底物。最近，许多研究人员表明，人类诱导多能干细胞（iPS）细胞分化而来的细胞，如肝细胞和肠细胞，可用于药代动力学测试。在这里，我们尝试使用基因组编辑技术生成 MDR1 敲除（KO）iPS 细胞系。成功获得了正确靶向的人类 iPS 细胞系。MDR1 敲除并未改变人 iPS 细胞中多能标志物的表达水平。MDR1-KO iPS 衍生的肝样细胞中肝标志物的基因表达水平高于未分化的 MDR1-KO iPS 细胞，表明 MDR1-KO iPS 细胞具有肝分化能力。此外，MDR1-KO iPS 细胞衍生的肝样细胞中几乎检测不到 MDR1 的表达。因此，我们成功建立了可用于药代动力学测试的 MDR1-KO iPS 细胞系。
