The 12-HETE receptor Gpr31 in the -cell pathogenesis of type 1 diabetes

摘要 本应用程序响应RFA-RM-19-011，并将重点放在 阐明了糖尿病背景下被称为GPR31的可药物基因组(IDG)数据库。超过了 在过去的十年里，我们的实验室一直专注于炎症和细胞对炎症的反应作为一种 导致β细胞功能障碍和死亡的中枢机制--1型糖尿病(T1D)和2型糖尿病 (T2D)。具体地说，我们的实验室已经证明了12-脂氧合酶(12-LOX)，一种参与 花生四烯酸代谢并在β细胞中表达，在β细胞炎症和 产生二十烷类化合物12-S-羟基二十碳四烯酸(12(S)-HETE)。12(S)-HETE产生 β细胞中的内质网(ER)和氧化应激，但其发生的机制有 仍然难以捉摸。小鼠12-LOX编码基因的缺失(Alox15)，无论是在胰岛还是在 在全球范围内，导致保护NOD小鼠免受自发性1型糖尿病和肥胖诱导的2型糖尿病的影响。 就像高脂饮食的小鼠患糖尿病一样。GPR31最近被解除孤儿，并被确定为12(S)-HETE 受体。我们通过传统的ES在C57BL6/J背景上产生了Gpr31b-/-小鼠 细胞克隆。初步数据表明，GPR31-/-小鼠是存活的，代谢正常，类似于ALOX15- /-小鼠，但GPR31是否介导12-LOX在糖尿病发生中的作用仍有待确定 β细胞中的条件。在这项提案中，我们将生成支持GPR31潜在作用的初步数据 作为炎症诱导的β细胞功能障碍和死亡的关键介质，以及产生的数据和资源 将向联合国儿童基金会资源传播和外联中心(RDoC)提供这份提案的 IDG程序。这项拨款提案延续了S·S博士之间长期而富有成效的合作。 特西和R·米尔米拉，他们共同位于芝加哥大学。博尔顿博士的综合专业知识。 β细胞信号级联中的米尔米拉和特西博士在糖尿病动物模型中的协同作用 完成这一试点提案。我们的团队将检验GPR31促进β-细胞炎症的假设 在糖尿病的背景下，信号转导和促进β细胞功能障碍和死亡。为了检验这一假设， 将在分配给本论坛的时间框架内实现以下两个目标： 目的1：阐明GPR31在糖尿病炎症和12(S)-HETE中的作用。 小岛。 目的：研究GPR31在正常和促炎条件下的体内代谢效应。 这项提议的主要影响是确定GPR31是否为合适的药物靶点 在糖尿病炎症背景下的发展。

Abstract This application is responsive to RFA-RM-19-011 and will focus on a poorly characterized GPCR in the Illuminating the Druggable Genome (IDG) database known as GPR31 in the context of diabetes. Over the past decade, our laboratory has focused on inflammation and the cellular response to inflammation as a central mechanism that contributes to β-cell dysfunction and death type 1 diabetes (T1D) and type 2 diabetes (T2D). Specifically, our laboratory has demonstrated that 12-lipoxygenase (12-LOX), an enzyme involved in arachidonic acid metabolism and expressed in β-cells, is activated in the context of β-cell inflammation and produces the eicosanoid 12-S-hydroxyeicosatetraenoic acid (12(S)-HETE). 12(S)-HETE generates endoplasmic reticulum (ER) and oxidative stress in β cells, but the mechanism through which this occurs has remained elusive. Deletion of the gene encoding 12-LOX in mice (Alox15), either conditionally in islets or globally, leads to protection from spontaneous type 1-like diabetes in NOD mice and obesity-induced type 2- like diabetes in high fat-fed mice. GPR31 has recently been de-orphaned and identified as the 12(S)-HETE receptor. We have generated congenic Gpr31b-/- mice on the C57BL6/J background through traditional ES cell cloning. Preliminary data suggest that Gpr31-/- mice are viable and metabolically normal, similar to Alox15- /- mice, however it remains to be determined if GPR31 mediates the effects of 12-LOX under diabetogenic conditions in β-cells. In this proposal, we will generate preliminary data supporting the potential role of GPR31 as a key mediator of inflammation-induced β-cell dysfunction and death, and the data and resources generated from this proposal will be made available to the Resource Dissemination and Outreach Center (RDOC) of the IDG program. This grant proposal continues a longstanding and productive collaboration between Drs. S. Tersey and R. Mirmira, who are co-located at the University of Chicago. The combined expertise of Dr. Mirmira in β cell signaling cascades and Dr. Tersey in animal models of diabetes are synergistic towards the completion of this pilot proposal. Our Team will test the hypothesis that GPR31 promotes β-cell inflammatory signaling and contributes to β-cell dysfunction and death in the setting of diabetes. To test this hypothesis, the following two aims will be achieved within the timeframe allotted to this RFA: Aim 1: Elucidate the role of GPR31 in mediating the effects of diabetogenic inflammation and 12(S)-HETE in islets. Aim 2: Characterize the metabolic effects of GPR31 in vivo under normal and pro-inflammatory conditions. The primary impact of this proposal is the determination of whether GPR31 is a suitable target for drug development in the context of diabetic inflammation.