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: 10877390
• 负责人: Christine Anne Klemens
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10877390
• 关键词: ATP phosphohydrolase; Acetylcholine; Address; Adult; Affect; Animal Experimentation; 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; Compensation; Contracts; Coupling; Dahl Hypertensive Rats; Data; Deposition; Development; Diet; Electrophysiology (science); Endoplasmic Reticulum; Equilibrium; Extracellular Matrix; Extracellular Matrix Proteins; Fibrosis; 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; 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; Postdoctoral Fellow; Process; Property; Protein Secretion; Proteins; Protocols documentation; Rattus; Research; Resistance; Risk; Role; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Sodium Chloride; Stimulus; Stroke; 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; mRNA Expression; migration; new therapeutic target; normotensive; novel; prevent; programs; protein expression; protein function; protein transport; reconstitution; response; skills; socioeconomics; stroke risk; therapeutic target; 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口患有高血压，这使他们处于中风的风险增加 损害，心脏病发作，心力衰竭和肾脏疾病。最近的全基因组关联研究已关联 基因CLCN6中的许多突变，可降低高血压和中风风险。 CLCN6编码 电压敏感的氯化物通道6（CLC-6）。迄今为止，关于功能CLC-6或其作用的知之甚少 在血压稳态中。该建议的总体目标是建立生理和分子 CLC-6在血管平滑肌细胞（VSMC）功能和血压中的作用。 指导阶段：初步数据证明CLC-6在高尔基体中表达 VSMC。我假设VSMC中的CLC-6活性通过提供A来调节Luminal Golgi Ca2+商店 Ca2+摄取的充电平衡，从而保持膜电压。这是通过 与高尔基体特异性Ca2+ -ATPase，SPCA1和CLC-6的损失的关联减少Golgi Ca2+商店 和信号，它改变了VSMC收缩性。具体目标1。确定CLC-6在Golgi Ca2+上的作用 在VSMC中处理。我将利用平面脂质双层电生理学和复杂的Ca2+成像 确定CLC-6氯化物通道性质及其对高尔基体CA2+存储容量的影响。 此外，这些实验将提供第一个证据，证明Golgi特异性Ca2+在VSMC中的作用 Ca2+对血管结构和扩张刺激的处理，从未被检查过。 独立阶段：该项目的这一阶段将开发一条独立的调查线 CLC-6对VSMC功能的分子效应。从这些实验中获得的洞察力将进一步解释 CLC-6调节血压控制的生理机制。我的初步数据 确定CLC-6可防止或减慢高血压发育过程中大型动脉血管的变化。我会 进一步研究CLC-6对细胞增殖，迁移，凋亡和细胞外基质蛋白的影响 在正常和高血压血管中沉积。我假设CLC-6的损失将减少 细胞外基质蛋白分泌和缓慢的细胞增殖，从而减少高血压诱导 纤维化和培养基增厚。这些变化将导致动脉僵硬和血管 重塑并降低周围血管耐药性，为CLC-6调节的方式提供了理由 血压。具体目标2。阐明CLC-6对血管刚度和血管重塑的影响 在高血压期间。解决此假设的实验将包括各种实验范围， 包括脉冲波速度测量，组织学分析，迁移分析，myhotemhing，隧道和 BRDU分析和转录组分析。这些研究将使我们对蛋白质功能的理解有所了解 通过在分子，细胞和全身水平上定义该通道在脉管系统中的作用。