权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The function of kidney specific (KS)-WNK1 condensates during potassium stress

钾应激期间肾脏特异性 (KS)-WNK1 缩合物的功能

基本信息

批准号：
10397071
负责人：
Cary Ragan Boyd-Shiwarski
金额：
$ 16.25万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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：
2018-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10397071
关键词：
Advisory Committees Affect Alanine Animal Model Animals Binding Proteins Biological Biology Blood Blood Pressure Chronic Kidney Failure Clustered Regularly Interspaced Short Palindromic Repeats Complex Data Development Diet Dietary Potassium Disease Distal convoluted renal tubule structure Duct (organ) structure Epidemic Equilibrium Evolution Foundations Funding Goals Homeostasis Human Hypertension Hypokalemia In Vitro Intake International Kidney Kidney Diseases Knockout Mice Lead Link Lysine Maps Mass Spectrum Analysis Measures Membrane Mentors Metabolic Microscopy Molecular Molecular Cloning Monitor Morbidity - disease rate Mus Mutagenesis Mutation Organelles Pathway interactions Phosphorylation Phosphotransferases Physicians Physiological Physiology Plasma Population Potassium Process Proline Protein Biochemistry Protein Isoforms Protein Kinase Proteins RNA Recommendation Regulation Renal function Research Research Personnel Resources RiboTag Ribosomes Role SLC12A3 gene Scientist Serum Signal Pathway Signal Transduction Slice Sodium Sodium Chloride Stress Structure Techniques Telemetry Testing Time Tissues Transgenic Mice United States National Institutes of Health Universities Urine Work base blood pressure elevation blood pressure regulation career development cost experience gain of function mutation human model hyperkalemia in vivo insight meetings mouse model novel pressure proteotoxicity renal tubular transport response sensor skills symporter tenure track thiazide transcriptome sequencing

项目摘要

PROJECT SUMMARY/ABSTRACT Potassium [K+]-deficient diets have contributed to the global epidemic of hypertension and chronic kidney disease (CKD). Given the low cost and ease of increasing dietary [K+], more research is needed to understand how [K+] imbalance leads to these diseases. The kidneys handle 90% of [K+], and the distal convoluted tubule (DCT) acts as a [K+] sensor via the WNK-SPAK (With-No-Lysine/Ste20/SPS-1-related proline-alanine-rich protein kinase) pathway. Gain-of-function mutations to this pathway lead to severe hypertension and hyperkalemia by activation of the thiazide-sensitive sodium/chloride co-transporter (NCC). Curiously, dietary [K+] depletion or loading causes the WNK-SPAK kinases to assemble into large DCT-specific cytoplasmic puncta, that are not seen in mice on normokalemic diets. For years, the structure and function of these condensates, which we call “WNK bodies”, remained a mystery. Dr. Boyd-Shiwarski’s initial work has identified that these DCT- specific puncta are (i) dependent upon the expression of kidney specific (KS)-WNK1 (ii) potassium-sensitive;; (iii) membrane-less;; (iv) not associated with conventional organelles;; and (v) associated with WNK-SPAK proteins. Based on these findings, we hypothesize that WNK bodies are membrane-less microdomains that sequester the WNK-SPAK pathway to modulate WNK signaling during potassium imbalance. This hypothesis will be tested in two aims that evaluate the physiological significance and biological basis of WNK body formation. This proposal’s physiology-based aim will provide Dr. Boyd-Shiwarski with the opportunity to work with animal models and (i) implement ex vivo microscopy techniques, (ii) quantify changes in urine, serum, and blood pressure, and (iii) develop transgenic mouse models. Whereas, the biology-based portion of this proposal will include implementation of (i) molecular cloning, (ii) protein biochemistry, and (iii) mass spectrometry and RNA Seq. These skills will be reinforced by a team of mentors, advisors, collaborators, and core resources available at the University of Pittsburgh. The primary mentor, Dr. Arohan Subramanya, is an established NIH R01-funded physician-scientist with 13 years of experience in WNK signaling biology and prior experience mentoring over 20 trainees. The co-mentor, Dr. Tom Kleyman, is an internationally recognized physician scientist who directs the Pittsburgh Center for Kidney Research, and has mentored nine career development awardees and five R01 recipients within the last 10 years. In addition, an advisory committee of accomplished investigators with expertise in hypertension, WNK-SPAK signaling, and renal tubular transport will monitor Dr. Boyd-Shiwarski’s progress through biannual meetings. Dr. Boyd-Shiwarski will use this proposal to accomplish her short-term goal of scientific independence and her long-term goal of becoming a tenure track physician-scientist with expertise in potassium homeostasis, hypertension, and CKD. The results from this proposal will form the basis for an R01 studying the translational role of KS-WNK1-dependent WNK bodies in human models of nephropathy.

项目总结/文摘