Stresser David M, Sun Jun, Wilson Sarah S
AbbVie, Inc., North Chicago, Illinois (D.M.S., J.S.) and AbbVie Cambridge Research Center, Cambridge, Massachusetts (S.S.W.)
AbbVie, Inc., North Chicago, Illinois (D.M.S., J.S.) and AbbVie Cambridge Research Center, Cambridge, Massachusetts (S.S.W.).
Drug Metab Dispos. 2021 Mar;49(3):245-253. doi: 10.1124/dmd.120.000281. Epub 2020 Dec 18.
Induction of cytochrome P450 can cause drug-drug interactions and efficacy failure. Induction risk in liver and gut is typically inferred from experiments with plated hepatocytes. Organoids are physiologically relevant, multicellular structures originating from stem cells. Intestinal stem cell-derived organoids retain traits of normal gut physiology, such as an epithelial barrier and cellular diversity. Matched human enteroid and colonoid lines, generated from ileal and colon biopsies from two donors, were cultured in extracellular matrix for 3 days, followed by a single 48-hour treatment with rifampin, omeprazole, CITCO, and phenytoin at concentrations that induce target genes in hepatocytes. After treatment, mRNA was analyzed for induction of target genes. Rifampin induced CYP3A4; estimated EC and maximal fold induction were 3.75 µM and 8.96-fold, respectively, for ileal organoids and 1.40 µM and 11.3-fold, respectively, for colon organoids. Ileal, but not colon, organoids exhibited nifedipine oxidase activity, which was induced by rifampin up to 14-fold. The test compounds did not increase mRNA expression of CYP1A2, CYP2B6, multidrug resistance transporter 1 (P-glycoprotein), breast cancer resistance protein, and UDP-glucuronosyltransferase 1A1 in ileal organoids. Whereas omeprazole induced CYP3A4 (up to 5.3-fold, geometric mean, = 4 experiments), constitutive androstane receptor activators phenytoin and CITCO did not. Omeprazole failed to induce CYP1A2 mRNA but did induce CYP1A1 mRNA (up to 7.7-fold and 15-fold in ileal and colon organoids, respectively, = 4 experiments). Despite relatively high intra- and interexperimental variability, data suggest that the model yields induction responses that are distinct from hepatocytes and holds promise to enable evaluation of CYP1A1 and CYP3A4 induction in gut. SIGNIFICANCE STATEMENT: An adult intestinal stem cell-derived organoid model to test P450 induction in gut was evaluated. Testing several prototypical inducers for mRNA induction of P450 isoforms, UDP-glucuronosyltransferase 1A1, P-glycoprotein, and breast cancer resistance protein with both human colon and ileal organoids resulted in a range of responses, often distinct from those found in hepatocytes, indicating the potential for further development of this model as a physiologically relevant gut induction test system.
细胞色素P450的诱导可导致药物相互作用和疗效失败。肝脏和肠道中的诱导风险通常从平板培养的肝细胞实验中推断得出。类器官是源自干细胞的具有生理相关性的多细胞结构。肠道干细胞衍生的类器官保留了正常肠道生理的特征，如上皮屏障和细胞多样性。从两名供体的回肠和结肠活检组织中生成的匹配的人肠类器官和结肠类器官系在细胞外基质中培养3天，然后用利福平、奥美拉唑、CITCO和苯妥英进行单次48小时处理，处理浓度为可诱导肝细胞中靶基因的浓度。处理后，分析mRNA以检测靶基因的诱导情况。利福平诱导CYP3A4；回肠类器官的估计EC50和最大诱导倍数分别为3.75 μM和8.96倍，结肠类器官分别为1.40 μM和11.3倍。回肠类器官而非结肠类器官表现出硝苯地平氧化酶活性，该活性被利福平诱导高达14倍。测试化合物未增加回肠类器官中CYP1A2、CYP2B6、多药耐药转运蛋白1（P-糖蛋白）、乳腺癌耐药蛋白和尿苷二磷酸葡萄糖醛酸基转移酶1A1的mRNA表达。虽然奥美拉唑诱导了CYP3A4（高达5.3倍，几何平均值，n = 4次实验），但组成型雄烷受体激活剂苯妥英和CITCO未诱导。奥美拉唑未能诱导CYP1A2 mRNA，但诱导了CYP1A1 mRNA（回肠和结肠类器官中分别高达7.7倍和15倍，n = 4次实验）。尽管实验内和实验间存在相对较高的变异性，但数据表明该模型产生的诱导反应与肝细胞不同，有望用于评估肠道中CYP1A1和CYP3A4的诱导情况。重要性声明：评估了一种用于测试肠道中P450诱导的成人肠道干细胞衍生类器官模型。用人结肠和回肠类器官测试几种典型诱导剂对P450同工酶、尿苷二磷酸葡萄糖醛酸基转移酶1A1、P-糖蛋白和乳腺癌耐药蛋白mRNA诱导的情况，得到了一系列反应，这些反应通常与肝细胞中的反应不同，表明该模型作为一种具有生理相关性的肠道诱导测试系统具有进一步开发的潜力。
