The Role of OAT1 in Uremia

项目摘要/摘要 OAT1(SLC22A6)是典型的肾脏有机阴离子转运体。它负责运输 在许多药物中(例如，抗生素、利尿剂、抗病毒药物、非甾体抗炎药)和毒素(有机汞)。我们确定了 Transporter(AS NKT)；我们还首次发表了Oat1基因敲除(Oat1KO)和表型。我们最近的数据 表明OAT1在许多全身性和近端小管代谢过程中起着中心作用。燕麦1KO 代谢组学数据表明，OAT1可能是体内尿毒症毒素在肾脏积聚的关键转运体 肾功能不全和慢性肾脏疾病(CKD)。改变OAT1功能和/或表达可能会改善 肾功能受损状态下的代谢变化。我们假设，人的“真正”生理作用 OAT1是一个更大的代谢网络的中心组成部分，参与正常的生理和 病理生理过程，而OAT1对代谢性疾病(如：尿毒症 综合症)通过这个网络。我们将详细定义以OAT1为中心的网络。具体：1)我们将 通过转录和代谢重组构建以OAT1为中心的代谢网络 来自Oat1KO的代谢组学数据(使用所述的系统生物学技术)。我们应该能够 清楚地描绘了OAT1通过以下方式调节正常代谢的途径：a)覆盖基于配体的 关于这个初步重建的以OAT1为中心的网络的药效团模型(手头)，以及b)测试 在湿法实验室运输化验中的“命中率”。2)然后我们将确定经验证的以OAT1为中心的角色 代谢网络(及其内部的关键通路)在肾功能不全的代谢改变中发挥作用。 啮齿动物肾大部切除模型和人类CKD代谢组学数据。我们寻求建立一个非常详细的， 经过充分验证的&gt；200代谢物和反应网络，做出可在以下方面评估的预测 病理生理模型和疾病状态。为回应先前的批评，我们修订了 提案，特别是在疾病模型方面--包括来自加州大学圣地亚哥分校的一名专家合作者 奥布莱恩中心协助研究和解读。尽管这个项目被认为雄心勃勃，但我们 强调大量的初步数据和“内部”专业知识。调查小组已经 在运输、代谢、网络生物学、肾脏疾病流行病学、动物模型和 大型数据集的统计分析。该项目将因此产生一张以OAT1为中心的经过验证的详细地图 在正常生理和疾病状态下的新陈代谢，可能是任何多特效药物的首例。 运输者(Nigam，《自然评论药物发现》，2015)。这些研究可能会导致制定战略 通过影响OAT1的功能或表达来改善CKD的代谢异常；它们也将有助于 预测OAT1转运药物如何影响肾脏疾病的新陈代谢。

PROJECT SUMMARY/ABSTRACT OAT1 (SLC22A6) is the prototypical kidney organic anion (PAH) transporter. It is responsible for the transport of many drugs (eg. antibiotics, diuretics, antivirals, NSAIDs) and toxins (organic mercurials). We identified the transporter (as NKT); we also first published the Oat1 knockout (Oat1KO) and phenotypes. Our recent data indicates that OAT1 is central to many systemic and proximal tubule metabolic processes. Oat1KO metabolomics data indicates OAT1 may be the key in vivo transporter of uremic toxins accumulating in renal insufficiency and chronic kidney disease (CKD). Altering OAT1 function and/or expression may ameliorate the metabolic changes in states of compromised renal function. We hypothesize that the “true” physiological role of OAT1 is as a central component of a larger metabolic network involved in normal physiological and pathophysiological processes, and OAT1 exerts a modulatory effect on metabolic diseases (eg. uremic syndrome) via this network. We will define the OAT1-centered network in detail. Specifically: 1) We will construct the OAT1-centered metabolic network through metabolic reconstruction of transcriptomics and metabolomics data from the Oat1KO (using the systems biology techniques described). We should be able to clearly delineate the pathways by which OAT1 regulates normal metabolism by: a) overlaying ligand-based pharmacophore models (in-hand) upon this preliminary reconstructed OAT1-centered network, and b) testing of “hits” in wet lab transport assays. 2) We will then determine what role the validated OAT1-centered metabolic network (and key pathways within it) plays in the metabolic alterations of renal insufficiency using rodent subtotal nephrectomy models and human CKD metabolomics data. We seek to build a very detailed, fully validated, network of >200 metabolites and reactions that makes predictions that can be evaluated in pathophysiological models and disease states. In response to previous criticisms, we have revised the proposal, particularly with respect to the disease model—and included an expert collaborator from the UCSD O’Brien Center to help with the studies and interpretation. Although the project was considered ambitious, we emphasize the huge amount of preliminary data and “in house” expertise. The team of investigators has expertise in transport, metabolism, network biology, epidemiology of kidney disease, animal models, and statistical analysis of large data sets. This project will thus produce a validated detailed map of OAT1-centered metabolism in normal physiology and a diseased state, possibly the first of its kind for any multispecific “drug” transporter (Nigam, Nature Reviews Drug Discovery, 2015). The studies could lead to design of strategies for improving the metabolic abnormalities in CKD by affecting OAT1 function or expression; they will also help predict how OAT1-transported drugs affect metabolism in kidney disease.