Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; PRESTO, Japan Science and Technology Agency, Saitama, Japan; Laboratory of Hepatocyte Regulation, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, Japan; Laboratory of Biochemistry and Molecular Biology, School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Cell Mol Gastroenterol Hepatol. 2019;8(3):513-526. doi: 10.1016/j.jcmgh.2019.06.004. Epub 2019 Jun 19.
BACKGROUND & AIMS: To develop an effective and safe orally administered drug, it is important to predict its intestinal absorption rate, intestinal first-pass effect, and drug-drug interactions of orally administered drugs. However, there is no existing model to comprehensively predict the intestinal pharmacokinetics and drug-response of orally administered drugs. In this study, we attempted to generate homogenous and functional intestinal epithelial cells from human induced pluripotent stem (iPS) cells for pharmaceutical research.
We generated almost-homogenous Villin- and zonula occludens-1 (ZO1)-positive intestinal epithelial cells by caudal-related homeobox transcription factor 2 (CDX2) transduction into human iPS cell-derived intestinal progenitor cells.
The drug absorption rates in human iPS cell-derived intestinal epithelial cell monolayers (iPS-IECM) were highly correlated with those in humans (R=0.91). The expression levels of cytochrome P450 (CYP) 3A4, a dominant drug-metabolizing enzyme in the small intestine, in human iPS-IECM were similar to those in human small intestine in vivo. In addition, intestinal availability in human iPS-IECM (the fraction passing the gut wall: Fg=0.73) was more similar to that in the human small intestine in vivo (Fg=0.57) than to that in Caco-2 cells (Fg=0.99), a human colorectal adenocarcinoma cell line. Moreover, the drug-drug interaction and drug-food interaction could be observed by using our human iPS-IECM in the presence of an inducer and inhibitor of CYP3A4, i.e., rifampicin and grape fruit juice, respectively.
Taking these results together, we succeeded in generating the human iPS-IECM that can be applied to various intestinal pharmacokinetics and drug-response tests of orally administered drugs.
为开发有效且安全的口服药物，预测其肠道吸收率、肠道首过效应和口服药物的药物相互作用非常重要。然而，目前还没有现有的模型可以全面预测口服药物的肠道药代动力学和药物反应。在本研究中，我们试图从人诱导多能干细胞（iPS）中生成同质且功能正常的肠上皮细胞，用于药物研究。
我们通过尾侧相关同源盒转录因子 2（CDX2）转导，将人 iPS 细胞衍生的肠祖细胞转化为几乎同质的绒毛蛋白（Villin）和紧密连接蛋白 1（ZO1）阳性肠上皮细胞。
人 iPS 细胞衍生的肠上皮细胞单层（iPS-IECM）中的药物吸收速率与人高度相关（R=0.91）。人 iPS-IECM 中细胞色素 P450（CYP）3A4 的表达水平，一种在小肠中占主导地位的药物代谢酶，与体内人小肠相似。此外，人 iPS-IECM 的肠道利用率（穿过肠壁的部分：Fg=0.73）与人小肠体内（Fg=0.57）更为相似，而与人类结肠直肠腺癌细胞系 Caco-2 细胞（Fg=0.99）相比则更相似。此外，在 CYP3A4 的诱导剂和抑制剂（利福平、葡萄柚汁）存在的情况下，我们可以用人 iPS-IECM 观察到药物-药物相互作用和药物-食物相互作用。
综合这些结果，我们成功生成了可应用于各种口服药物肠道药代动力学和药物反应测试的人 iPS-IECM。
