Role of the renal organic anion transporter OAT1 in metabolism and physiology

项目概要/摘要 由于新的监管指南，PI 发现的有机阴离子转运蛋白 (OAT1) 团体（如 NKT）并参与消除许多常见药物 - 已收到相当多的 注意。但几乎所有已发表的研究都继续关注常见药物而不是内源性药物 转运蛋白的底物，从进化的角度来看是高度保守的。代谢组学 对我们的 Oat1 敲除小鼠 (Oat1KO) 的分析以及体外转运数据表明，OAT1 是一种 代谢物、信号分子、维生素、肠道微生物组产品的主要转运蛋白 抗氧化剂。我们之前对 Oat1KO 代谢改变的基因组规模代谢重建 正常饮食条件下的小鼠使我们能够创建第一个多特异性“药物”转运蛋白（OAT1）- 中心代谢网络——大量代谢组学和体外数据进一步支持了这一点。 许多途径（例如脂肪酸、肠道微生物组产物、支链氨基酸）在很大程度上 经验证的约 150-200 种代谢物网络也被认为在代谢疾病中很重要，例如 糖尿病、代谢综合征、肥胖和肠道微生物相关疾病。在本提案中：1）我们将使用 Oat1 KO 小鼠的饮食调整（例如高脂肪、支链氨基酸）和抗生素治疗 定义 Oat1 在适用于代谢疾病的条件下的作用。我们将测试绑定 代谢组学揭示了不同饮食条件（或疾病模型）下的代谢物，使用高 通过未标记通量 (BLI) 测定来量化代谢物与 OAT1 的相互作用；该信息将是 用于对涉及使用放射性标记化合物的后续摄取测定中的代谢物进行优先排序。 2） 然后，我们将使用这些信息，以及来自不同条件下的基因敲除组织的基因表达数据。 饮食条件，为每种饮食（病理模型）条件重建代谢网络- -使用我们之前成功创建“以OAT1为中心的代谢网络”的方法 正常饮食条件。这些研究将共同帮助确定 OAT1 在监管中的独特作用 这些失调的代谢状态下的异常代谢。最后，我们讨论这些信息将如何 为了解服用 OAT1- 的患者药物引起的代谢综合征的各个方面奠定基础 运输毒品。我们在必要的技术方面拥有经过验证的专业知识和世界一流的团队 合作者以确保成功。

PROJECT SUMMARY/ABSTRACT Owing to new regulatory guidelines, the organic anion transporter (OAT1)--a transporter discovered by the PI's group (as NKT) and which is involved in the elimination of many common drugs--has received considerable attention. But nearly all the published studies continue to focus on common drugs rather than endogenous substrates of the transporter, which is highly conserved from an evolutionary standpoint. Metabolomics analysis of our Oat1 knockout mice (Oat1KO), as well as in vitro transport data, indicate that OAT1 is a major transporter of metabolites, signaling molecules, vitamins, gut microbiome products, and antioxidants. Our previous genome-scale metabolic reconstructions of altered metabolism of the Oat1KO mouse under normal dietary conditions enabled us to create the first multispecific "drug" transporter (OAT1)- centered metabolic network--which was further supported by considerable metabolomics and in vitro data. Many pathways (e.g., fatty acids, gut microbiome products, branched chain amino acids) in this largely validated network of ~150-200 metabolites are also known to be important in metabolic diseases such as diabetes, metabolic syndrome, obesity, and gut microbiome-associated illness. In this proposal: 1) We will use dietary modifications (e.g., high fat, branched chain amino acids) and antibiotic treatment of the Oat1 KO mice to define the role of Oat1 under conditions applicable to metabolic diseases. We will test the binding of metabolites revealed by metabolomics under the various dietary conditions (or disease models) using a high throughput unlabeled (BLI) assay to quantify the interaction of metabolites with OAT1; this information will be used to prioritize metabolites for subsequent uptake assays involving the use of radiolabeled compounds. 2) We will then use this information, together with gene expression data from the knockout tissues under various dietary conditions, to reconstruct a metabolic network for each of these dietary (pathological model) conditions- -using methods we have previously successfully used to create an "OAT1-centered metabolic network" under normal dietary conditions. Together, these studies will help define the unique roles of OAT1 in regulation of aberrant metabolism in these dysregulated metabolic states. Finally, we discuss how this information will set the stage for understanding aspects of drug-induced metabolic syndrome in patients taking OAT1- transported drugs. We have proven expertise in the necessary techniques and a team of world-class collaborators to ensure success.