Role of Nedd4-2, SGK1, and 14-3-3 in ENaC Trafficking

描述（由申请人提供）： 高血压（HTN）影响发达国家25%的成年人口，是中风、心肌梗死和充血性心力衰竭的主要独立危险因素。导致HTN的几种单基因缺陷现在已被归因于醛固酮敏感性远端肾单位内钠处理的异常，其中上皮Na+通道ENaC构成肾脏中Na+转运的限速步骤。因此，阐明ENaC调控机制将对HTN的研究具有深远的病理生理意义。这项研究将重点关注Nedd 4 - 2、SGK 1和14-3-3蛋白在ENaC运输中的作用。本实验的目的是：（1）研究Nedd 4 -2调控ENaC转运的分子机制。(2)确定与Nedd 4 - 2相互作用并抑制Nedd 4 -2的特异性14-3-3同种型的亚细胞定位。(3)评价SGK 1和14-3-3调节Nedd 4 -2的分子机制。 除了拟议的实验，教学课程在细胞生物学，共聚焦显微镜，和研究的负责任的行为将补充一个结构化的导师培训计划，以准备主要研究者（PI）的学术生涯作为一个临床科学家在肾脏病。大卫皮尔斯博士是离子运输激素调节领域的领导者，他将指导Pi的科学发展。此外，基思莫斯托夫博士是上皮细胞胞内运输领域的领导者，将在实验设计和结果解释方面提供科学专业知识。一个由有成就的生物医学科学家组成的专业发展咨询委员会也将定期召开会议，提供科学和职业建议。UC San弗朗西斯科提供的多样化科学资源和结构化指导是培训临床科学家的理想选择。在该培训项目和环境中，PI将发展成为一名有竞争力的独立研究者。

DESCRIPTION (provided by applicant): Hypertension (HTN) affects 25% of the adult population in the developed world and is a major, independent risk factor for stroke, myocardial infarction, and congestive heart failure. Several monogenetic defects resulting in HTN have now been attributed to abnormalities in sodium handling within the aldosterone-sensitive distal nephron where the epithelial Na+ channel, ENaC, constitutes the rate-limiting step of Na+ transport in the kidney. Therefore, elucidating the mechanisms of ENaC regulation will have profound pathophysiologic implications for the study of HTN. The proposed research will focus on the role of Nedd4- 2, SGK1, and 14-3-3 proteins in ENaC trafficking. The proposed experiments aim to: (1) Characterize the molecilar mechanisms by which Nedd4-2 regulates trafficking of ENaC. (2) Determine the subcellular localization of the specific 14-3-3 isoform(s) which interact with and inhibit Nedd4-2. (3) Evaluate the molecular mechanism of SGK1 and 14-3-3 regulation of Nedd4-2. In addition to the proposed experiments, didactic coursework in cell biology, confocal microscopy, and the responsible conduct of research will complement a structured mentorship training program to prepare the principal investigator (PI) for an academic career as a clinical scientist in Nephrology. Dr. David Pearce is a leader in the field of hormone regulation of ion transport and will mentor the Pi's scientific development. Additionally, Dr. Keith Mostov is a leader in the field of intracellular trafficking in epithelial cells and will provide scientific expertise in the design of experiments and interpretation of results. A professional development advisory committee comprised of accomplished biomedical scientists will also convene at regular intervals to provide scientific and career advice. The diverse, scientific resources and structured mentoring available at UC San Francisco are ideal for training clinical scientists. Within this training program and environment the PI will develop into a competitive, independent investigator.