Investigating the role of NBCe1-B in renal and cardiac acid-base handling

研究 NBCe1-B 在肾脏和心脏酸碱处理中的作用

基本信息

批准号：
10450763
负责人：
Clayton Timothy Brady
金额：
$ 3.25万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-07 至 2025-08-06
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10450763
关键词：
Acid-Base Equilibrium Acidosis Acids Address Adult Amino Acid Sequence Ammonia Animals Bicarbonates Blood Circulation Blood gas Body Weight Buffaloes Cardiac Cardiac Myocytes Cardiac development Cardiology Cardiovascular Diseases Catheterization Cause of Death Cells Chronic Chronic Kidney Failure Clinical Clinical Trials Data Development Disease Disease Progression Echocardiography Electrocardiogram Evaluation Excretory function Fellowship Functional disorder Goals Growth Heart Heart Hypertrophy Heart failure Histologic Homeostasis Immunohistochemistry Impairment Investigation Ion Transport Kidney Kidney Diseases Knock-out Knockout Mice Knowledge Laboratories Left Link Measurement Mentorship Metabolic acidosis Microscopy Modeling Molecular Mus Nephrology Outcome Patients Phenotype Physicians Physiologic intraventricular pressure Population Preceptorship Promoter Regions Proximal Kidney Tubules Regulation Renal function Research Respiratory Acidosis Risk Factors Role Science Scientist Series Serum System Testing Training Universities Variant Western Blotting Wild Type Mouse Work base experience experimental study heart function in vivo insight medical schools mortality pressure prevent response symporter

项目摘要

Project Summary An estimated 15% of U.S. adults are estimated to have chronic kidney disease (CKD), with cardiovascular disease (CVD) the greatest cause of death in this population. Metabolic acidosis is common in CKD, and has deleterious effects on both the kidney and the heart. Complicating our understanding and limiting our ability to prevent these effects are gaps in our knowledge of the integrated cardiorenal response to acidosis, which is largely influenced by cellular level acid-base transporters. The Na+/HCO3- co-transporter (NBCe1-B) is one such transporter. There is accumulating evidence that NBCe1-B is involved in the renal response to acidosis and the pathophysiologic development of cardiac hypertrophy, but the consequence of NBCe1-B loss on these systems has never been tested at the whole animal level. The goal of this project is to determine the role of NBCe1-B in the kidney and heart by comparing the renal and cardiac phenotypes of wild-type (WT) and NBCe1-B knockout (KO) mice. The over-arching hypothesis of this proposal is that NBCe1-B is essential for renal (Aim 1) and cardiac (Aim 2) acid-base handling. The purpose of Aim 1 is to determine the abundance, distribution, and function of NBCe1-B in the WT mouse kidney during acid-challenged conditions. This work will include a series of western-blot and fluorescent immunohistochemistry experiments as well as a comparison of renal acid-base handling during acidosis in WT and NBCe1-B KO mice as assessed by blood-gas parameters, ammonia excretion, and titratable acid excretion. The purpose of Aim 2 is to determine the exact type and mechanism of cardiac impairment in NBCe1-B-KO mice. The mice will receive a cardiac work-up similar to patients with heart- failure, incorporating electrocardiogram, echocardiogram, and left-ventricular pressure catheterization measurements. Cellular level investigation of cardiomyocytes will include histological analysis, molecular assessment for evidence of pro-hypertrophic mechanisms using Western blot and RT-qPCR, and evaluation of Ca2+ handling in isolated cardiomyocytes using Ca2+ sensitive microscopy. The experiments outlined in this proposal will determine the role of NBCe1-B in the renal and cardiac systems, providing insights into the cardiorenal response to acidosis. This work will take place at the University at Buffalo, Jacobs School of Medicine and Biomedical Sciences (JSMBS), in the laboratory of Dr. Mark Parker, who is an expert in ion transport and pH regulation. The training plan is tailored for development as a physician-scientist in the field of nephrology, and will include clinical preceptorships in nephrology and cardiology in order to gain cross-disciplinary experience, reflecting the research goals of this proposal. These longitudinal clinical preceptorships will be with successful physician-scientists, who will also be directly involved in the proposed research, thereby providing integrated mentorship over the course of the fellowship.

项目概要据估计，15% 的美国成年人患有慢性肾脏病 (CKD)，其中包括心血管疾病疾病（CVD）是该人群死亡的最大原因。代谢性酸中毒在 CKD 中很常见，并且对肾脏和心脏都有有害影响。使我们的理解变得复杂并限制了我们的能力预防这些影响是我们对酸中毒的综合心肾反应知识的空白，这是很大程度上受细胞水平酸碱转运蛋白的影响。 Na+/HCO3- 协同转运蛋白 (NBCe1-B) 就是这样的一种运输者。越来越多的证据表明 NBCe1-B 参与肾脏对酸中毒的反应和心脏肥大的病理生理学发展，以及 NBCe1-B 缺失对这些系统的影响从未在整个动物水平上进行过测试。该项目的目标是确定 NBCe1-B 在通过比较野生型 (WT) 和 NBCe1-B 敲除的肾脏和心脏表型来观察肾脏和心脏（KO）小鼠。该提案的首要假设是 NBCe1-B 对于肾脏（目标 1）和心脏（目标 2）酸碱处理。目标 1 的目的是确定丰度、分布和在酸挑战条件下 NBCe1-B 在 WT 小鼠肾脏中的功能。这部作品将包括一个系列蛋白质印迹和荧光免疫组织化学实验以及肾脏酸碱比较通过血气参数、氨评估评估 WT 和 NBCe1-B KO 小鼠酸中毒期间的处理排泄和可滴定酸排泄。目标 2 的目的是确定确切的类型和机制 NBCe1-B-KO 小鼠的心脏损伤。这些小鼠将接受与心脏病患者类似的心脏检查失败，包括心电图、超声心动图和左心室压力导管插入术测量。心肌细胞的细胞水平研究将包括组织学分析、分子分析使用蛋白质印迹和 RT-qPCR 评估促肥大机制的证据，并评估使用 Ca2+ 敏感显微镜对分离的心肌细胞进行 Ca2+ 处理。本文概述的实验该提案将确定 NBCe1-B 在肾脏和心脏系统中的作用，提供对心肾对酸中毒的反应。这项工作将在布法罗大学雅各布医学院进行和生物医学科学 (JSMBS)，在 Mark Parker 博士的实验室中，他是离子传输和生物医学领域的专家 pH调节。该培训计划是为肾脏病领域的医师科学家的发展而量身定制的，并将包括肾脏病学和心脏病学的临床指导，以获得跨学科的知识经验，反映了本提案的研究目标。这些纵向临床指导将与成功的医师科学家，他们也将直接参与拟议的研究，从而提供整个研究金过程中的综合指导。