Drug Metabolism & Pharmacokinetics, Research Laboratory for Development, SHIONOGI & CO., LTD., 3-1-1, Toyonaka, 561-0825, Osaka, Japan.
Department of Life Science and Technology, School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, 226-8501, Kanagawa, Japan.
Sci Rep. 2020 Apr 6;10(1):5989. doi: 10.1038/s41598-020-63151-z.
Drug absorption via the intestinal tissue is modulated by membrane permeability and metabolism in intestinal epithelial cells (IECs). In drug discovery research, using human IECs to evaluate membrane permeability and metabolic stability can offer very useful information when exploring for drug candidate compounds that have good bioavailability and when trying to predict the fraction absorbed and intestinal availability in humans. Here, we evaluated the pharmacokinetic functions of human IECs differentiated from human induced pluripotent stem cells (hiPSCs) in 3D cultures. As human IECs differentiated in 3D cultures form intestinal organoids and spheroids (herein termed organoids), their morphology makes it difficult to evaluate their pharmacokinetic functions. Therefore, we dissociated intestinal organoids into single cells and attempted to purify human IECs. We found that hiPSC-derived IECs (hiPSC-IECs) expressed the epithelial cell adhesion molecule (EpCAM) and could be highly purified by sorting EpCAM+ cells. The hiPSC-IEC monolayer showed a high TEER value (approximately 350 Ω × cm). In addition, hiPSC-IECs oxidatively metabolized terfenadine (CYP3A and CYP2J2 substrate) and midazolam (CYP3A substrate). These results indicated that hiPSC-IECs form tight-junction and have cytochrome P450 enzymatic activities. In conclusion, we developed a novel application of hiPSC-derived intestinal organoids for drug testing.
药物通过肠道组织的吸收受到肠道上皮细胞（IEC）中膜通透性和代谢的调节。在药物发现研究中，使用人源 IEC 评估膜通透性和代谢稳定性，在探索具有良好生物利用度的候选药物化合物以及尝试预测人体吸收分数和肠道利用率时，可以提供非常有用的信息。在这里，我们评估了 3D 培养中分化的人诱导多能干细胞（hiPSC）来源的人 IEC 的药代动力学功能。由于 3D 培养中分化的人 IEC 形成肠类器官和球体（本文称为类器官），它们的形态使得难以评估其药代动力学功能。因此，我们将肠类器官解离成单细胞，并尝试纯化人 IEC。我们发现，hiPSC 衍生的 IEC（hiPSC-IEC）表达上皮细胞黏附分子（EpCAM），并可以通过分选 EpCAM+细胞进行高度纯化。hiPSC-IEC 单层的 TEER 值很高（约 350 Ω×cm）。此外，hiPSC-IEC 氧化代谢特非那定（CYP3A 和 CYP2J2 底物）和咪达唑仑（CYP3A 底物）。这些结果表明，hiPSC-IEC 形成紧密连接并具有细胞色素 P450 酶活性。总之，我们开发了一种新型的 hiPSC 衍生肠类器官用于药物测试的应用。
