Discovering and dissecting new regulators of insulin production in beta cells

描述（由申请人提供）：由于我们无法阻止这种潜在疾病的发展，全球糖尿病患病率的空前上升更加令人担忧。我的职业目标是指导一个专注于开发新的糖尿病治疗方法的基础研究项目。使用RNA干扰（RNAi）筛选，我已经确定了一种新的G蛋白偶联受体（GPCR），它对胰岛素的产生和分泌很重要。目的一：明确该新型GPCR对胰岛素启动子和胰岛素分泌作用的分子机制。我将确定该GPCR对胰岛素启动子和胰岛素分泌的影响需要哪些G蛋白信号通路。我还将确定转录因子和胰岛素启动子中的调节位点它们是由这个GPCR调节的。使用Gq/11偶联工程GPCR的平行方法也将进行。目的二世。确定该GPCR在体内β细胞发育和胰岛素产生中的作用。我将研究这种GPCR的敲除小鼠，重点是β细胞发育和成人β细胞功能。第三目标。确定胰岛素启动子活性所需的新基因。扩展用于识别Aim I中确定的候选RNAi筛选，我将使用高通量系统筛选针对所有已知基因的sirna，以寻找胰岛素启动子的其他调节因子。这些目标将为我第一次申请R01提供必要的培训和独立研究方向。具体来说，我将学习使用糖尿病的小鼠模型和全基因组RNAi筛选的设计和性能。为了获得这种经验，我加入了Michael McManus博士的实验室，他是RNA干扰筛选和非编码RNA方面的专家，以及紧邻的Michael German博士的实验室，他是一位在体外和体内研究胰腺β细胞的内科科学家。我还有两位优秀的合作者——GPCR信号专家Bruce Conklin博士和UCSF高通量筛选设施负责人Michelle Arkin博士。加州大学旧金山分校和加州大学旧金山分校糖尿病中心为年轻研究人员提供了极好的培训环境。我将利用他们丰富的资源。为了协助第三目标，我将参加UCSF生物医学信息学项目的大数据集统计分析课程。为了职业发展，我还将参加有关负责任的研究行为、拨款写作和科学写作的课程和研讨会。我将参加一些关于糖尿病、RNAi或非编码rna的系列研讨会和联合实验室会议，并介绍我的研究。重要的是，我在UCSF的职位允许我100%的精力投入到我的训练中。综上所述，我建议研究一种新的GPCR在胰岛素产生中的机制，并确定其他在β细胞功能中重要的新基因。这笔资金不仅可以让我完成这些研究，还可以让我成为一名独立的糖尿病研究者。

DESCRIPTION (provided by applicant): The unprecedented rise in worldwide diabetes prevalence is made even more alarming by our inability to halt the progression of this insidious disease. My career goal is to direct a basic research program focused on developing new diabetes therapies. Using an RNA interference (RNAi) screen, I have identified a novel G- protein coupled receptor (GPCR) that is important for insulin production and secretion. My aims are as follows: Aim I. Define the molecular mechanism of this novel GPCR's action on the insulin promoter and insulin secretion. I will identify what G protein signaling pathway(s) are required for this GPCR's effect on the insulin promoter and insulin secretion. I will also identify the transcription factors and the regulatory sites in the insulin promoter that are regulated by this GPCR. A parallel approach using a Gq/11-coupled engineered GPCR will also be undertaken. Aim II. Determine the role of this GPCR in beta cell development and insulin production in vivo. I will study a knockout mouse for this GPCR focusing on beta cell development and adult beta cell function. Aim III. Identify novel genes required for insulin promoter activity. Expanding on the RNAi screen used to identify the candidate identified in Aim I, I will use a high throughput system to screen siRNAs targeting all known genes to find additional regulators of the insulin promoter. These aims will provide the training and independent research direction necessary for my first R01 application. Specifically, I will learn to use mouse models of diabetes and the design and performance of whole genome RNAi screens. To gain this experience, I have joined the laboratory of Dr. Michael McManus, an expert on RNA interference screening and non-coding RNAs, and the immediately adjacent laboratory of Dr. Michael German, a physician scientist who studies the pancreatic beta cell both in vitro and in vivo. I also have two excellent collaborators -- Dr. Bruce Conklin, an expert in GPCR signaling, and Dr. Michelle Arkin, the leader of the UCSF High Throughput Screening facility. UCSF and the UCSF Diabetes Center are excellent training environments for young investigators. I will capitalize on their rich resources. To assist with Aim III, I will enroll in the UCSF Biomedical Informatics Program's class on the statistical analysis of large data sets. For career development, I will also participate in classes and workshops on the responsible conduct of research, grant writing, and scientific writing. I will attend and present my research in several seminar series and joint lab meetings focused on diabetes, RNAi or non- coding RNAs. Importantly, my position at UCSF allows 100% of my effort to be devoted to my training. In summary, I propose to study the mechanism of a novel GPCR in insulin production and to identify other novel genes important in beta cell function. This grant will not only allow the completion of these studies but will also allow me to transition into an independent diabetes investigator. PUBLIC HEALTH RELEVANCE: Diabetes or pre-diabetes affects nearly 40% adults of in the US. Current treatments for diabetes are inadequate. The goal of this proposal is the identification of genes required for insulin production with the hope that these genes will be drug targets for the next generation of diabetes therapies.