The Role of CIC-6 in Vascular Control of Blood Pressure

CIC-6 在血管血压控制中的作用

• 批准号: 10460452
• 负责人: Christine Anne Klemens
• 金额: $ 10.06万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-18 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10460452
• 关键词: ATP phosphohydrolase; Acetylcholine; Address; Adult; Affect; Animal Model; Apoptosis; Apoptotic; Applications Grants; Arteries; Biochemical; Biological Assay; Biological Models; Blood Pressure; Blood Vessels; Bromodeoxyuridine; Ca(2+)-Transporting ATPase; Calcium; Cardiac; Carrier Proteins; Cell Proliferation; Cell physiology; Cellular biology; Charge; Chloride Channels; Chlorides; Code; Contracts; Coupling; Dahl Hypertensive Rats; Data; Deposition; Development; Electrophysiology (science); Endoplasmic Reticulum; Equilibrium; Extracellular Matrix; Extracellular Matrix Proteins; Fibrosis; Financial compensation; Foundations; Future; Genes; Goals; Golgi Apparatus; Heart failure; Histologic; Histology; Human; Hypertension; Image; In Situ Nick-End Labeling; In Vitro; Injury to Kidney; Investigation; Ions; Kidney Diseases; Knock-out; Link; Lipid Bilayers; Measurement; Measures; Medial; Membrane; Mentors; Migration Assay; Mitochondria; Molecular; Molecular Biology; Muscle Contraction; Mutation; Myocardial Infarction; Myography; Organelles; Pathologic; Peripheral; Phase; Phenotype; Physiologic pulse; Physiological; Play; Population; Post-Translational Protein Processing; Process; Property; Protein Secretion; Protocols documentation; Rattus; Research; Resistance; Risk; Role; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Sodium Chloride; Stimulus; Therapeutic; Translating; Variant; Vascular Smooth Muscle; Vascular resistance; Work; arterial stiffness; biophysical properties; blood pressure control; blood pressure reduction; blood pressure regulation; burden of illness; economic impact; experimental study; genome wide association study; health economics; hypertensive; insight; kidney vascular structure; mRNA Expression; migration; new therapeutic target; normotensive; novel; prevent; programs; protein expression; protein function; protein transport; reconstitution; response; salt sensitive; skills; socioeconomics; stroke risk; transcriptomics; uptake; voltage; wound healing

PROJECT SUMMARY Nearly half of the US adult population has hypertension, which puts them at increased risk for stroke, vascular damage, heart attack, heart failure, and kidney disease. Recent genome-wide association studies have linked a number of mutations in the gene, CLCN6, to reduced hypertension and stroke risk. CLCN6 encodes the voltage-sensitive chloride channel 6 (ClC-6). To date, very little is known of about the function ClC-6, or its role in blood pressure homeostasis. The overall goal of this proposal is to establish the physiological and molecular roles of ClC-6 on vascular smooth muscle cell (VSMC) function and blood pressure. Mentored Phase: Preliminary data have demonstrated that ClC-6 is expressed in the Golgi apparatus of VSMCs. I hypothesize that ClC-6 activity in VSMCs regulates luminal Golgi Ca2+ stores by providing a charge balance for Ca2+ uptake, thereby maintaining membrane electroneutrality. This occurs through an association with the Golgi-specific Ca2+-ATPase, SPCA1, and loss of ClC-6 reduces Golgi Ca2+ stores and signaling, which alters VSMC contractility. Specific Aim 1. Establish the role of ClC-6 on Golgi Ca2+ handling in VSMCs. I will utilize planar lipid bilayer electrophysiology and sophisticated Ca2+ imaging to determine ClC-6 chloride channel properties and their impact on the Ca2+ storage capacity of the Golgi. Furthermore, these experiments will provide the first evidence of the role of Golgi-specific Ca2+ release in VSMC Ca2+ handling in response to vasocontraction and dilation stimuli, which has never before been examined. Independent Phase: This phase of the project will develop an independent line of investigation into the molecular effects of ClC-6 on VSMC function. Insight gained from these experiments will further explain the physiological mechanism underlying ClC-6 regulation of blood pressure control. My preliminary data have established that ClC-6 prevents or slows large artery vessel stiffening during development of hypertension. I will further examine the effect of ClC-6 on cellular proliferation, migration, apoptosis, and extracellular matrix protein deposition in normotensive and hypertensive vessels. I hypothesize that loss of ClC-6 will reduce extracellular matrix protein secretion and slow cell proliferation, thereby reducing hypertension induced fibrosis and media thickening. These changes will result in abrogated arterial stiffening and vessel remodeling, and reduce peripheral vascular resistance, providing a rationale for how ClC-6 moderates blood pressure. Specific Aim 2. Elucidate the influence of ClC-6 on vascular stiffness and vessel remodeling during hypertension. Experiments to address this hypothesis will include a diverse range of experiments, including pulse-wave velocity measurements, histological analyses, migration assays, myography, TUNNEL and BrdU assays, and transcriptomic analysis. These studies will inform our understanding of the protein's function by defining the role of this channel in vasculature at the molecular, cellular, and systemic levels.

项目摘要 近一半的美国成年人患有高血压，这使他们患中风、血管性心脏病和高血压的风险增加。 心脏病心脏衰竭和肾脏疾病最近的全基因组关联研究表明， CLCN 6基因的一些突变，以降低高血压和中风的风险。CLCN 6编码 电压敏感性氯离子通道6（ClC-6）。迄今为止，对ClC-6的功能或其作用知之甚少 在血压稳态方面。该提案的总体目标是建立生理和分子生物学模型， ClC-6对血管平滑肌细胞（VSMC）功能和血压的作用。 指导阶段：初步数据表明，ClC-6在高尔基体中表达。 的VSMC。我假设VSMCs中ClC-6活性通过提供一个钙通道来调节腔高尔基体的钙库。 Ca 2+摄取的电荷平衡，从而维持膜电中性。这是通过 与高尔基体特异性Ca 2 +-ATP酶、SPCA 1和ClC-6缺失的关联减少了高尔基体的Ca 2+储存 和信号传导，其改变VSMC收缩性。具体目标1.确定ClC-6对高尔基体Ca 2+的作用 在VSMC中处理。我将利用平面脂质双层电生理学和复杂的钙离子成像， 确定ClC-6氯离子通道特性及其对高尔基体的Ca 2+储存能力的影响。 此外，这些实验将提供高尔基体特异性Ca 2+释放在VSMC中的作用的第一个证据 Ca 2+处理血管收缩和扩张刺激，这是以前从未被检查。 独立阶段：该项目的这一阶段将制定一个独立的调查路线， ClC-6对VSMC功能的分子作用。从这些实验中获得的见解将进一步解释 ClC-6调节血压控制的生理机制。我的初步数据显示 C1 C-6预防或减缓高血压发展过程中大动脉血管硬化。我会 进一步检测ClC-6对细胞增殖、迁移、凋亡和细胞外基质蛋白的影响， 在正常血压和高血压血管中沉积。我假设ClC-6的丢失会减少 细胞外基质蛋白的分泌和细胞增殖缓慢，从而降低高血压诱导的 纤维化和中膜增厚。这些变化将导致废除动脉硬化和血管 重塑，并降低外周血管阻力，为ClC-6如何调节 血压.具体目标2。阐明ClC-6对血管僵硬度和血管重塑的影响 在高血压期间。解决这一假设的实验将包括各种各样的实验， 包括脉搏波速度测量、组织学分析、迁移测定、肌造影、隧道和 BrdU测定和转录组学分析。这些研究将为我们了解蛋白质的功能提供信息 通过在分子、细胞和系统水平上定义该通道在脉管系统中的作用。