Department of Pharmacology, College of Medicine, University of Arizona, Tucson, Arizona
Department of Pharmacology, College of Medicine, University of Arizona, Tucson, Arizona.
J Pharmacol Exp Ther. 2021 Feb;376(2):148-160. doi: 10.1124/jpet.120.000267. Epub 2020 Nov 9.
Our in vivo rodent studies have shown that organic anion transporting polypeptide (Oatp) 1a4 is critical for blood-to-brain transport of statins, drugs that are effective neuroprotectants. Additionally, transforming growth factor- (TGF-) signaling via the activin receptor-like kinase 1 (ALK1) receptor regulates Oatp1a4 functional expression. The human ortholog of Oatp1a4 is OATP1A2. Therefore, the translational significance of our work requires demonstration that OATP1A2 can transport statins and is regulated by TGF-/ALK1 signaling. Cellular uptake and monolayer permeability of atorvastatin, pravastatin, and rosuvastatin were investigated in vitro using human umbilical vein endothelial cells (HUVECs). Regulation of OATP1A2 by the TGF-/ALK1 pathway was evaluated using bone morphogenetic protein 9 (BMP-9), a selective ALK1 agonist, and LDN193189, an ALK1 antagonist. We showed that statin accumulation in HUVECs requires OATP1A2-mediated uptake but is also affected by efflux transporters (i.e., P-glycoprotein, breast cancer resistance protein). Absorptive flux (i.e., apical-to-basolateral) for all statins was higher than secretory flux (i.e., basolateral-to-apical) and was decreased by an OATP inhibitor (i.e., estrone-3-sulfate). OATP1A2 protein expression, statin uptake, and cellular monolayer permeability were increased by BMP-9 treatment. This effect was attenuated in the presence of LDN193189. Apical-to-basolateral statin transport across human endothelial cellular monolayers requires functional expression of OATP1A2, which can be controlled by therapeutically targeting TGF-/ALK1 signaling. Taken together with our previous work, the present data show that OATP-mediated drug transport is a critical mechanism in facilitating neuroprotective drug disposition across endothelial barriers of the blood-brain barrier. SIGNIFICANCE STATEMENT: Transporter data derived from rodent models requires validation in human models. Using human umbilical vein endothelial cells, this study has shown that statin transport is mediated by OATP1A2. Additionally, we demonstrated that OATP1A2 is regulated by transforming growth factor-/activin receptor-like kinase 1 signaling. This work emphasizes the need to consider endothelial transporter kinetics and regulation during preclinical drug development. Furthermore, our forward-thinking approach can identify effective therapeutics for diseases for which drug development has been challenging (i.e., neurological diseases).
我们的体内啮齿动物研究表明，有机阴离子转运多肽 (Oatp) 1a4 对于他汀类药物的血脑转运至关重要，而他汀类药物是有效的神经保护剂。此外，通过激活素受体样激酶 1 (ALK1) 受体的转化生长因子- (TGF-) 信号调节 Oatp1a4 的功能表达。Oatp1a4 的人类同源物是 OATP1A2。因此，我们工作的转化意义需要证明 OATP1A2 可以转运他汀类药物，并且受到 TGF-/ALK1 信号的调节。使用人脐静脉内皮细胞 (HUVEC) 在体外研究阿托伐他汀、普伐他汀和罗苏伐他汀的细胞摄取和单层渗透性。使用骨形态发生蛋白 9 (BMP-9)（一种选择性 ALK1 激动剂）和 LDN193189（ALK1 拮抗剂）评估 TGF-/ALK1 途径对 OATP1A2 的调节。我们表明，他汀类药物在 HUVEC 中的积累需要 OATP1A2 介导的摄取，但也受到外排转运蛋白（即 P-糖蛋白、乳腺癌耐药蛋白）的影响。所有他汀类药物的吸收通量（即，从顶端到基底外侧）均高于分泌通量（即，基底外侧到顶端），并且被 OATP 抑制剂（即雌酮-3-硫酸盐）降低。BMP-9 处理增加了 OATP1A2 蛋白表达、他汀类药物摄取和细胞单层渗透性。在用 LDN193189 存在的情况下，这种作用减弱。穿过人内皮细胞单层的他汀类药物从顶端到基底外侧的转运需要 OATP1A2 的功能表达，而通过治疗靶向 TGF-/ALK1 信号可以控制这种表达。结合我们之前的工作，本数据表明 OATP 介导的药物转运是促进神经保护药物穿过血脑屏障内皮屏障的关键机制。意义声明：从啮齿动物模型中得出的转运体数据需要在人体模型中进行验证。使用人脐静脉内皮细胞，本研究表明他汀类药物的转运由 OATP1A2 介导。此外，我们证明了 OATP1A2 受转化生长因子-/激活素受体样激酶 1 信号的调节。这项工作强调了在临床前药物开发过程中需要考虑内皮转运体动力学和调节。此外，我们的前瞻性方法可以确定对药物开发具有挑战性的疾病（即神经疾病）的有效治疗方法。
