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

• 批准号: 10224674
• 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
• 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/10224674
• 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; 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

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.

项目摘要/摘要 维持水稳态是肾脏的重要功能，对于适应地面至关重要 生活。为了有效地吸收水，加压素（VP）释放以诱导水通道蛋白-2（AQP2）磷酸化 和肌动蛋白细胞骨架重塑集合管中的肾脏主要细胞中，这增加了顶端 AQP2的膜表达。 AQP2贩运的失调导致水平疾病； AQP2膜表达降低会导致肾脏基糖尿病（NDI），而增加 质膜AQP2与不适当ADH分泌综合征的液体保留有关 （SIADH），充血性心力衰竭和肝硬化。而VP/CAMP/PKA是主要的信号通路 促进了AQP2膜运输和吸水，该过程实际上更为复杂，并且可以 被其他信号通路诱导或抑制。这样的“替代”途径涉及表皮 生长因子受体（EGFR），其抑制作用诱导AQP2膜积累和磷酸化 类似于VP，但绕过V2R，CAMP和PKA。因此，这笔赠款的目的是1）定义 VP和EGFR信号通路之间的串扰机制，该途径调节AQP2运输； 2） 了解AQP2磷酸化在细胞骨架重塑中的作用，3）表征 在充血性心力衰竭患者中观察到的水位失调的新型信号通路，在 为了最终设计疗法以减轻诊所遇到的疾病症状。 培训计划是进步的，旨在将Pi Dr. Cheung博士穿越初始阶段 完成并扩大她在EGF/VP Crosstalk上正在进行的令人兴奋的工作（AIM 1），然后转到新的 在实验室（AIM 2）中，AQP2细胞骨架相互作用的方面（AIM 2）最终到达AIM 3之前 在心脏/肾脏互动和水平上，这将完成她的独立之路，并成为 她的第一个R01申请的主题。由她的精神布朗博士监督的张博士不仅会雇用 膜生物学计划中的现有技术，但也将获得重要的新技术和 研究培训都来自她的个性化培训计划中列出的课程，以及她的专家 咨询委员会Drs。 Joung（CRISPR/CAS9基因编辑）和Nahrendorf（手术和鼠标成像 心脏衰竭）。对于职业职业发展，除了获得屡获殊荣的导师布朗博士之外，她还 有博士。西尔维·布雷顿（Sylvie Breton）和乔迪·巴比特（Jodie Babitt）作为成功的女性学术科学家榜样。而且， MGH肾脏科负责人Ravi Thadhani博士将保护她的研究时间免受临床职责，允许 完全沉浸在她的研究中。她的精神，她的科学咨询委员会的成员，她的部门负责人， MGH和哈佛社区的培养与协作环境将与仔细的 设计的科学计划旨在推动她成为独立R01资助的学术的目标 肾脏科医生具有参与水调节的管间通信方面的专业知识。