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; 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 估计有15％的美国成年人患有慢性肾脏疾病（CKD），心血管疾病 疾病（CVD）该人群中最大的死亡原因。代谢性酸中毒在CKD中很常见，并且具有 对肾脏和心脏的有害影响。使我们的理解变得复杂，并限制了我们的能力 防止这些影响是我们对酸中毒综合反应的综合反应的差距，即 在很大程度上受细胞水平酸碱转运蛋白的影响。 NA+/HCO3-共转运蛋白（NBCE1-B）就是这样一种 转运蛋白。有积极的证据表明NBCE1-B参与了对酸中毒和 心脏肥大的病理生理发展，但这些系统上NBCE1-B丢失的结果 从未在整个动物层面进行过测试。该项目的目的是确定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进行肾脏和心脏 （KO）小鼠。该提议的总体假设是NBCE1-B对于肾脏至关重要（AIM 1）和 心脏（AIM 2）酸碱处理。目标1的目的是确定丰度，分布和 在酸挑战的条件下，NBCE1-B在WT小鼠肾脏中的功能。这项工作将包括一个系列 西部印迹和荧光免疫组织化学实验以及肾酸基碱的比较 在WT和NBCE1-B KO小鼠中酸中毒期间的处理，如血液气体参数评估 排泄和可滴定酸排泄。目标2的目的是确定确切类型和机制 NBCE1-B-KO小鼠的心脏障碍。小鼠将接受类似于心脏患者的心脏检查 故障，包含心电图，超声心动图和左心压导管插入术 测量。心肌细胞的细胞水平研究将包括组织学分析，分子 评估使用Western blot和RT-QPCR的促嗜性机制的证据，并评估 使用Ca2+敏感显微镜在分离的心肌细胞中进行CA2+处理。在此概述的实验 提案将确定NBCE1-B在肾脏和心脏系统中的作用，从而提供见解 对酸中毒的心脏反应。这项工作将在雅各布斯医学院的布法罗大学举行 和生物医学科学（JSMBS），在Mark Parker博士的实验室中，他是离子运输专家 pH调节。该培训计划是针对肾脏学领域的医师科学家开发而定制的， 并将包括肾脏学和心脏病学领域的临床主持，以获得跨学科 经验，反映该提案的研究目标。这些纵向的临床教育将与 成功的医师科学家也将直接参与拟议的研究，从而提供 在研究金的过程中综合指导。