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

Nedd4-2、SGK1 和 14-3-3 在 ENaC 贩运中的作用

• 批准号: 7274319
• 负责人: VIVEK BHALLA
• 金额: $ 12.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7274319
• 关键词: 14-3-3 Proteins; Acute; Adult; Advisory Committees; Affect; Aldosterone; Apical; Biological Assay; Blood Pressure; Cell Line; Cell membrane; Cell surface; Cells; Cellular biology; Clathrin; Clinical; Complement; Condition; Confocal Microscopy; Congestive Heart Failure; Cultured Cells; Data; Defect; Development; Distal; Duct (organ) structure; Electrolytes; Endocytosis; Environment; Epithelial Cells; Family; Hypertension; In Vitro; Ion Channel; Ion Transport; Kidney; Label; Laboratories; Ligase; Lysine; Maintenance; Mediating; Membrane Protein Traffic; Mentors; Mentorship; Microscopy; Molecular; Monitor; Mus; Myocardial Infarction; Nephrology; Nephrons; Numbers; Phosphorylation; Physiological; Play; Polymerase Chain Reaction; Population; Principal Investigator; Process; Protein Isoforms; Proteins; RNA Interference; Rate; Regulation; Research; Research Personnel; Resources; Risk Factors; Role; San Francisco; Scientist; Sgk protein; Sodium; Sorting - Cell Movement; Stroke; Structure; Surface; System; Testing; Training; Training Programs; Transcript; base; biomedical scientist; career; design; epithelial Na+ channel; genetic regulatory protein; hormone regulation; inhibitor/antagonist; member; novel therapeutics; programs; research study; trafficking; ubiquitin ligase

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.

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、心肌梗塞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已归因于醛固酮敏感的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 调节机制将对 HTN 的研究具有深远的病理生理学意义。拟议的研究将重点关注 Nedd4-2、SGK1 和 14-3-3 蛋白在 ENaC 运输中的作用。拟议的实验旨在：（1）表征 Nedd4-2 调节 ENaC 运输的分子机制。 (2) 确定与 Nedd4-2 相互作用并抑制 Nedd4-2 的特定 14-3-3 同工型的亚细胞定位。 (3)评价SGK1和14-3-3调控Nedd4-2的分子机制。 除了拟议的实验之外，细胞生物学、共焦显微镜和负责任的研究行为方面的教学课程将补充结构化的指导培训计划，为首席研究员（PI）作为肾病学临床科学家的学术生涯做好准备。 David Pearce 博士是离子传输激素调节领域的领导者，他将指导 Pi 的科学发展。此外，Keith Mostov 博士是上皮细胞细胞内运输领域的领导者，并将在实验设计和结果解释方面提供科学专业知识。由杰出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。加州大学旧金山分校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。在这个培训计划和环境中，PI 将发展成为一名有竞争力的独立调查员。