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

• 批准号: 10669150
• 负责人: Clayton Timothy Brady
• 金额: $ 5.27万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 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/10669150
• 关键词: Acid-Base Equilibrium; Acidosis; Acids; Address; Adult; Amino Acid Sequence; Ammonia; Animals; Bicarbonates; Blood gas; Body Weight; Cardiac; Cardiac Myocytes; Cardiology; Cardiovascular Diseases; Catheterization; Cause of Death; Cells; Chronic; Chronic Kidney Failure; Circulation; 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; absorption; base; experience; experimental study; heart function; in vivo; insight; knowledge integration; medical schools; mortality; pressure; prevent; response; symporter

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.

项目摘要 据估计，15%的美国成年人患有慢性肾病（CKD），伴有心血管疾病。 心血管疾病（CVD）是这一人群的最大死因。代谢性酸中毒在CKD中很常见， 对肾脏和心脏的有害影响。使我们的理解复杂化，限制了我们的能力， 防止这些影响是我们对酸中毒的综合心肾反应知识的空白， 很大程度上受细胞水平酸碱转运蛋白的影响。Na+/HCO 3-共转运蛋白（NBCe 1-B）就是这样一种 传送器。越来越多的证据表明，NBCe 1-B参与了对酸中毒的肾反应， 心肌肥大的病理生理发展，但NBCe 1-B损失对这些系统的后果 从未在整个动物水平上进行过测试。该项目的目标是确定NBCe 1-B在 通过比较野生型（WT）和NBCe 1-B敲除的肾脏和心脏表型， (KO)小鼠该建议的过度假设是NBCe 1-B对于肾（目的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 在酸攻击条件下WT小鼠肾脏中NBCe 1-B的功能。这项工作将包括一系列 免疫印迹和荧光免疫组化实验以及肾酸碱平衡的比较 WT和NBCe 1-B KO小鼠酸中毒期间的处理，通过血气参数、氨 排泄和可滴定酸排泄。目标2的目的是确定 NBCe 1-B-KO小鼠的心脏损伤。小鼠将接受类似于心脏病患者的心脏检查- 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、分子生物学分析、细胞学分析和细胞生物学分析。 使用蛋白质印迹和RT-qPCR评估促肥大机制的证据， 使用Ca 2+敏感显微镜在分离的心肌细胞中的Ca 2+处理。本文中概述的实验 该提案将确定NBCe 1-B在肾脏和心脏系统中的作用，为研究NBCe 1-B在肾脏和心脏系统中的作用提供见解。 对酸中毒的心肾反应。这项工作将在布法罗大学雅各布斯医学院进行 Mark帕克博士的实验室，他是离子传输和生物医学科学的专家， pH调节。培训计划是为发展成为肾脏病学领域的医生-科学家而量身定制的， 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 经验，反映了本提案的研究目标。这些纵向临床指导将与 成功的医生科学家，谁也将直接参与拟议的研究，从而提供 在研究金期间提供综合指导。