Discovering and dissecting new regulators of insulin production in beta cells

描述（由申请人提供）：全球糖尿病患病率前所未有的上升，由于我们无法阻止这种阴险疾病的进展而变得更加令人担忧。我的职业目标是指导一个专注于开发新的糖尿病疗法的基础研究项目。使用RNA干扰（RNAi）筛选，我已经确定了一个新的G-蛋白偶联受体（GPCR），这是重要的胰岛素的生产和分泌。我的目标如下：目标一。明确这种新型GPCR对胰岛素启动子和胰岛素分泌作用的分子机制。我将确定这种GPCR对胰岛素启动子和胰岛素分泌的作用所需的G蛋白信号通路。我还将确定转录因子和调控位点的胰岛素启动子，这是由该GPCR的调节。还将采用Gq/11偶联工程GPCR的平行方法。Aim II.确定这种GPCR在体内β细胞发育和胰岛素产生中的作用。我将研究基因敲除小鼠，专注于β细胞发育和成人β细胞功能。Aim III.确定胰岛素启动子活性所需的新基因。在用于鉴定目标I中鉴定的候选物的RNAi筛选的基础上，我将使用高通量系统来筛选靶向所有已知基因的siRNA，以找到胰岛素启动子的其他调节剂。这些目标将为我的第一个R 01申请提供必要的培训和独立的研究方向。具体来说，我将学习使用糖尿病小鼠模型和全基因组RNAi筛选的设计和性能。为了获得这方面的经验，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细胞的医生科学家。我还有两位优秀的合作者--布鲁斯·康克林博士，GPCR信号专家，和米歇尔·阿金博士，加州大学旧金山分校高通量筛选设施的负责人。UCSF和UCSF糖尿病中心是年轻研究人员的绝佳培训环境。我将利用他们丰富的资源。为了帮助实现目标III，我将报名参加UCSF生物医学信息学项目的大型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地进行研究，资助写作和科学写作的课程和研讨会。我将出席并在几个系列研讨会和联合实验室会议上介绍我的研究，重点是糖尿病，RNAi或非编码RNA。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。总之，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。这笔赠款不仅可以完成这些研究，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.