Characterization of Novel Signaling Pathways Involved in Water Balance Disorders

与水平衡紊乱相关的新型信号通路的表征

基本信息

批准号：
9979859
负责人：
Pui Wen Cheung
金额：
$ 17.28万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9979859
关键词：
AVPR2 gene Actins Advisory Committees Affect Anterior Arteries Attenuated Award Binding Biological Assay Bypass CRISPR/Cas technology Cardiac Cell Culture Techniques Cell Line Cell membrane Cells Cirrhosis Clinic Clinical Clustered Regularly Interspaced Short Palindromic Repeats Co-Immunoprecipitations Communication Communities Complex Congestive Heart Failure Coronary Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cytoskeleton Data Dehydration Disease Duct (organ) structure Epidermal Growth Factor Receptor Epithelial Epithelium Equilibrium Erlotinib Female Forskolin Functional disorder Funding Genes Goals Grant Growth Factor Guanosine Triphosphate Heart Heart failure Hepatotoxicity Homeostasis Hydrolysis Hyponatremia Immersion In Situ In Vitro Inappropriate ADH Syndrome Kidney Kidney Concentrating Ability Knockout Mice Left Life Ligation Liquid substance Lithium Liver Lung Magnetic Resonance Imaging Maintenance Measures Membrane Membrane Biology Mentors Metabolic Microscopy Modeling Mus Mutation Nephrology Operative Surgical Procedures Oral Organ Osmolalities Output Pathology Pathway interactions Patients Phosphorylation Phosphotransferases Play Point Mutation Process Proteins Pyrenes Receptor Inhibition Receptor Signaling Regulation Renal function Research Research Training Rhodamine Role Scientist Secondary to Serum Signal Pathway Signal Transduction Slice Small Interfering RNA Supervision Symptoms Techniques Technology Testing Time Training Urine Vasopressins Ventricular Water Western Blotting Work apical membrane aquaporin-2 career development cell growth regulation collaborative environment design equilibration disorder heart imaging immunocytochemistry in vitro activity in vivo inhibitor/antagonist kinase inhibitor knock-down member mutant novel polymerization programs receptor rho GTPase-activating protein role model targeted treatment therapy design tolvaptan trafficking vasopressin resistant diabetes insipidus water channel

项目摘要

PROJECT SUMMARY/ABSTRACT Maintenance of water homeostasis is a vital function of the kidneys and is essential for adaptation to terrestrial life. To reabsorb water effectively, vasopressin (VP) is released to induce aquaporin-2 (AQP2) phosphorylation and actin cytoskeletal remodeling within kidney principal cells in the collecting ducts, which increases apical membrane expression of AQP2. Dysregulation of AQP2 trafficking results in disorders of water balance; decreased AQP2 membrane expression causes nephrogenic diabetes insipidus (NDI), whereas an increase in plasma membrane AQP2 is associated with fluid retention in the syndrome of inappropriate ADH secretion (SIADH), congestive heart failure and cirrhosis. While VP/cAMP/PKA is the major signaling pathway that facilitates AQP2 membrane trafficking and water reabsorption, the process is in fact far more complex, and can be induced or inhibited by other signaling pathways. One such “alternative” pathway involves the epidermal growth factor receptor (EGFR), whose inhibition induces AQP2 membrane accumulation and phosphorylation similar to VP, but bypasses V2R, cAMP and PKA. Therefore, the aims of this grant are to 1) define the mechanism of crosstalk between VP and EGFR signaling pathways that regulate AQP2 trafficking; 2) understand the role of AQP2 phosphorylation on cytoskeletal remodeling, and 3) characterize the roles of novel signaling pathways in dysregulated water retention observed in patients with congestive heart failure, in order to eventually design therapies to alleviate disease symptoms encountered in the clinic. The training plan is progressive and is designed to move the PI Dr. Cheung through an initial stage of completing and expanding her exciting, ongoing work on EGF/VP crosstalk (Aim 1), then moving on to a new aspect of AQP2 cytoskeletal interactions not so far examined in the lab (Aim 2), before finally arriving at Aim 3 on heart/kidney interactions and water balance, which will complete her pathway to independence and be the subject of her first R01 application. Dr. Cheung, supervised by her mentor Dr. Brown, will not only employ existing technologies in the Program in Membrane Biology, but will also acquire important new techniques and research training both from courses listed in her personalized training plan, and from the experts on her advisory committee Drs. Joung (CRISPR/Cas9 gene editing) and Nahrendorf (surgery and mouse imaging for heart failure). For professional career development, in addition to her award-winning mentor Dr. Brown, she has Drs. Sylvie Breton and Jodie Babitt as successful female academic scientist role models. Moreover, the MGH Nephrology Division chief, Dr. Ravi Thadhani will protect her research time from clinical duties, allowing full immersion in her research. Her mentor, members of her scientific advisory committee, her Division chief, the nurturing and collaborative environment in the MGH and Harvard community, will combine with a carefully designed scientific plan to propel her toward her goal of becoming an independent R01 funded academic nephrologist with expertise in inter-organ communications involved in water regulation.

项目概要/摘要维持水稳态是肾脏的一项重要功能，对于适应陆地环境至关重要为了有效地重新吸收水分，加压素 (VP) 被释放以诱导水通道蛋白 2 (AQP2) 磷酸化。和集合管肾主细胞内的肌动蛋白细胞骨架重塑，这增加了顶端 AQP2 的膜表达失调导致水平衡紊乱； AQP2 膜表达会导致肾性尿崩症 (NDI)，而 AQP2 的增加会导致肾性尿崩症 (NDI) 质膜 AQP2 与 ADH 分泌不当综合征中的液体潴留相关 (SIADH)、充血性心力衰竭和肝硬化，而 VP/cAMP/PKA 是主要的信号通路。促进 AQP2 膜运输和水重吸收，该过程实际上要复杂得多，并且可以一种这样的“替代”途径涉及表皮。生长因子受体 (EGFR)，其抑制作用会诱导 AQP2 膜积聚和磷酸化与 VP 类似，但绕过 V2R、cAMP 和 PKA 因此，本次资助的目的是 1) 定义调节 AQP2 运输的 VP 和 EGFR 信号通路之间的串扰机制；了解 AQP2 磷酸化对细胞骨架重塑的作用，以及 3) 表征在充血性心力衰竭患者中观察到的水潴留失调的新信号通路为了最终设计出缓解临床中遇到的疾病症状的疗法。培训计划是渐进式的，旨在让 PI 张博士度过初始阶段：完成并扩展她正在进行的关于 EGF/VP 串扰的令人兴奋的工作（目标 1），然后转向新的在最终达到目标 3 之前，AQP2 细胞骨架相互作用的方面尚未在实验室中进行检查（目标 2）关于心/肾相互作用和水平衡，这将完成她的独立之路，并成为她的第一个R01申请的主题，在她的导师布朗博士的指导下，不仅会雇用。膜生物学计划中的现有技术，但还将获得重要的新技术和研究培训既来自她的个性化培训计划中列出的课程，也来自她的专家顾问委员会 Joung 博士（CRISPR/Cas9 基因编辑）和 Nahrendorf（手术和小鼠成像）对于职业生涯的发展，除了她屡获殊荣的导师布朗博士之外，她西尔维·布雷顿 (Sylvie Breton) 博士和朱迪·巴比特 (Jodie Babitt) 博士是成功的女性学术科学家的榜样。 MGH 肾脏病科主任 Ravi Thadhani 博士将保护她的研究时间，使其免受临床职责的影响，从而允许她的导师、她的科学顾问委员会成员、她的部门负责人、麻省总医院和哈佛社区的培育和协作环境，将与精心设计的项目相结合设计了科学计划来推动她实现成为独立 R01 学者资助的目标肾病专家，在涉及水调节的器官间通讯方面具有专业知识。