Novel Calcium Signaling Nanodomains in Vascular Smooth Muscle Cells

血管平滑肌细胞中的新型钙信号纳米结构域

• 批准号: 10744522
• 负责人: Swapnil K. Sonkusare
• 金额: $ 53.51万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10744522
• 关键词: A kinase anchoring protein; Adrenergic Receptor; Adult; Arteries; Biological Assay; Blood Pressure; Calcium Signaling; Caveolae; Cell membrane; Co-Immunoprecipitations; Contracts; Data; Dilator; Electrophysiology (science); Elements; Equilibrium; Hypertension; Impairment; Individual; Ion Channel; Knockout Mice; Ligation; Link; Measurement; Measures; Mediating; Molecular; Mus; Muscle Contraction; Myography; Nerve; PRKCA gene; Pathway interactions; Patients; Peptides; Piezo 1 ion channel; Play; Protein Kinase C; Proteins; Receptor Activation; Receptor Signaling; Reporting; Resistance; Rest; Role; Ryanodine Receptor Calcium Release Channel; Sarcoplasmic Reticulum; Scaffolding Protein; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Speed; Stimulus; Testing; Therapeutic; United States; Vanilloid; Vascular Smooth Muscle; Vascular remodeling; Vascular resistance; blood pressure control; blood pressure elevation; blood pressure reduction; blood pressure regulation; caveolin 1; cell type; clinically relevant; confocal imaging; design; experimental study; hypertensive; inhibitor; large-conductance calcium-activated potassium channels; mouse model; nano; new therapeutic target; novel; patch clamp; pressure; protein activation; receptor; receptors for activated C kinase; scaffold; superresolution imaging; vasoconstriction

Smooth muscle Ca2+ signaling mechanisms are crucial regulators of arterial contraction and blood pressure. Abnormalities in arterial smooth muscle cell (SMC) Ca2+ signaling mechanisms have been linked to vasoconstriction, vascular remodeling, and blood pressure elevation in hypertension. Therefore, identifying a new Ca2+ signaling mechanism could provide fresh impetus for designing novel therapeutic targets that lower blood pressure in hypertension. In this regard, TRPV4 (transient receptor potential vanilloid 4) ion channels are a well-known Ca2+-influx pathway in SMCs. Using smooth muscle-specific TRPV4-knockout mice, we provided first evidence that SMC TRPV4 channels increase resting blood pressure and contribute to blood pressure elevation in hypertension. Moreover, our preliminary data demonstrate two TRPV4 channel-containing signaling nanodomains in SMCs with opposite functional effects: (1) constrictor nanodomains involving nerve stimulation-induced activation of α1 adrenergic receptors (α1ARs)–protein kinase C (PKC)-anchoring protein AKAP150–TRPV4 channel signaling; and (2) dilator nanodomains activated by intraluminal pressure and comprising caveolin-1 scaffolded Piezo1–TRPV4–Ca2+- activated K+ (BK) channel signaling. Further, we show that constrictor α1AR–TRPV4 channel signaling is accentuated in arteries from a mouse model of hypertension and hypertensive patients, whereas dilator TRPV4–BK channel signaling is reduced. We hypothesize that miscommunication between the signaling elements disrupts the balance between constrictor and dilator signaling nanodomains and leads to blood pressure elevation in hypertension. In Aim 1, we will use smooth muscle-specific knockout mice, protein co-localization, patch-clamp electrophysiology, and blood pressure radiotelemetry to determine the molecular mechanisms underlying the excessive activation of constrictor α1AR:AKAP150:PKC:TRPV4 nanodomains in hypertension. In Aim 2, we will determine the molecular mechanisms responsible for the reduced activity of dilator Piezo1:TRPV4:BK channel nanodomains scaffolded by caveolin-1 in hypertension. The clinical relevance of studies in each Aim will be established by experiments in arteries from non-hypertensive and hypertensive individuals. Collectively, the proposed studies will establish novel SMC Ca2+-signaling nanodomains that control blood pressure and identify specific impairments at these nanodomains in hypertension.

平滑肌Ca2+信号传导机制是动脉收缩和血液的关键调节剂 压力。动脉平滑肌细胞（SMC）CA2+信号传导机制的异常 与血管收缩，血管重塑和血压升高有关 高血压。因此，确定新的CA2+信号传导机制可以提供新的动力 用于设计降低高血压血压的新型热靶标。在这方面， TRPV4（瞬态接收器电位香草素4）离子通道是众所周知的Ca2+-influx途径 在SMC中。使用平滑肌特异性TRPV4敲除小鼠，我们提供了首先证据表明 SMC TRPV4通道会增加静息血压并导致血压升高 在高血压中。此外，我们的初步数据证明了两个含TRPV4通道的数据 具有相反功能效应的SMC中的信号纳米域：（1）收缩纳米域 涉及神经刺激引起的α1肾上腺素受体（α1ARS） - 蛋白激酶的激活 C（PKC） - 锚定蛋白AKAP150 – TRPV4通道信号传导； （2）扩张纳米构域 被腔内压力并完成小窝蛋白-1脚手架的Piezo1 – Trpv4 – Ca2+ - 激活 激活的K+（BK）信号传导。此外，我们表明收缩α1AR– TRPV4通道 来自高血压小鼠模型和高血压的小鼠模型中的动脉突出信号传导 患者，而扩张器TRPV4 -BK通道信号传导降低。我们假设这一点 信号传导元素之间的沟通不畅破坏了收缩和之间的平衡 扩张器信号纳米域并导致高血压的血压升高。在AIM 1中， 我们将使用平滑肌特异性敲除小鼠，蛋白质共定位，贴片钳 电生理学和血压放射性电子链体，以确定分子机制 限制器α1AR的过量激活的基础：AKAP150：PKC：TRPV4纳米域中 高血压。在AIM 2中，我们将确定负责减少的分子机制 扩张器压电的活性：TRPV4：Caveolin-1在In In In caveolin-1脚手架的BK通道纳米域 高血压。每个目标中研究的临床相关性将通过实验建立 来自非高血压和高血压个体的动脉。拟议的研究集体 将建立新的SMC Ca2+信号纳米域，以控制血压并识别 高血压中这些纳米域的特定损伤。