Characterizing the physiological and pharmacological roles of SLC22A24

SLC22A24的生理和药理作用研究 溶质载体超家族(SLC)中的膜转运蛋白在生理学和生物化学中发挥着重要作用。 用于细胞摄取和清除内源性代谢物的药理学和许多方剂 毒品。尽管它们很重要，但大约五分之一的SLC转运蛋白没有已知的底物，而是 被认为是“孤儿”蛋白质。最近，我们的实验室通过以下方式使其中一种蛋白质SLC22A24去孤儿 应用各种方法，包括对全基因组关联信号的计算分析， 同源建模、表达谱分析，以及重要的是，稳定表达的细胞的同位素摄取分析 SLC22A24。我们的研究表明SLC22A24是一种对硫酸盐有偏好的有机阴离子转运体 和类固醇的葡萄糖醛酸化物结合物。最近的初步研究表明，转运蛋白还扮演着一个 在药物处置中的重要作用。拟议研究的主要目标是确定 SLC22A24在糖皮质激素代谢中的生理和药理作用 发现作为SLC22A24底物的其他内源性代谢物和药理制剂。 为了实现我们的目标，我们设计了三个具体目标。在目标1中，我们将利用细胞系的研究来 确定SLC22A24在基底膜或根尖膜上的定位。另外，我们会发现， 作为SLC22A24底物的处方药和其他代谢物。在目标2中，我们将使用CRISPR/Cas9 方法建立近端小管表达SLC22A24的转基因小鼠，并用其检测 它在转运蛋白的药理和生理底物的处理中的作用。最后，在目标3中，我们 我将在健康志愿者中进行关联研究，将SLC22A24的功能变异与血浆联系起来 类固醇葡萄糖醛酸苷的水平和肾脏清除量。使用的方法将包括共聚焦显微镜， 细胞的同位素摄取和通量分析，小鼠的CRISPR/Cas9方法，LC/MS/代谢组学分析 方法、关联分析和人体清除量测定。我们假定这一点 在细胞、小鼠和健康志愿者中进行的全面基因组、代谢和功能研究，包括 深入的临床表型将作为系统评估生理学和 一种新的去孤儿转运蛋白的药理学作用。

Characterizing the Physiological and Pharmacological Roles of SLC22A24 Membrane transporters in the solute carrier superfamily (SLC) play critical roles in physiology and pharmacology serving in the cellular uptake and removal of endogenous metabolites and many prescription drugs. In spite of their importance, about one-fifth of SLC transporters have no known substrates and are considered “orphan” proteins. Recently, our laboratory de-orphaned one of these proteins, SLC22A24, by applying various methods, which included computational analyses of genomewide association signals, homology modeling, expression profiling, and importantly, isotopic uptake assays in cells stably expressing SLC22A24. Our studies revealed that SLC22A24 is an organic anion transporter with a preference for sulfate and glucuronide conjugates of steroids. Recent preliminary studies suggest that the transporter also plays an important role in drug disposition. The major goals of the proposed research are to determine the physiological and pharmacological roles of SLC22A24 in the disposition of steroid glucuronides and to discover other endogenous metabolites and pharmacologic agents that are substrates of SLC22A24. To achieve our goals, we have designed three specific aims. In aim 1, we will employ studies in cell lines to determine the localization of SLC22A24 to the basolateral or apical membranes. In addition, we will discover prescription drugs and other metabolites that are substrates of SLC22A24. In aim 2, we will use CRISPR/Cas9 methods to create transgenic mice expressing SLC22A24 in the proximal tubule and use the mice to determine its role in the disposition of pharmacologic and physiologic substrates of the transporter. Finally, in aim 3, we will conduct association studies in healthy volunteers to associate functional variants in SLC22A24 with plasma levels and renal clearance of steroid glucuronides. Methods employed will include confocal microscopy, isotopic uptake and flux assays in cells, CRISPR/Cas9 methods in mice, LC/MS/ metabolomic analytical methods, association analyses and clearance determinations in humans. We postulate that this comprehensive genomic, metabolomic and functional studies in cells, mice and in healthy volunteers including deep clinical phenotyping will serve as a blueprint for systematic evaluation of the physiological and pharmacological roles of a newly de-orphaned transporter.