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重要调节因子 动脉平滑肌细胞 (SMC) Ca2+ 信号传导机制异常。 与血管收缩、血管重塑和血压升高有关 因此，确定一种新的 Ca2+ 信号传导机制可以提供新的动力。 设计降低高血压患者血压的新治疗靶点。 TRPV4（瞬时受体电位香草酸 4）离子通道是众所周知的 Ca2+ 流入途径 使用平滑肌特异性 TRPV4 敲除小鼠，我们提供了第一个证据： SMC TRPV4 通道会增加静息血压并导致血压升高 此外，我们的初步数据表明含有两个 TRPV4 通道。 SMC 中具有相反功能作用的信号纳米结构域：(1) 收缩纳米结构域 神经刺激诱导的 α1 肾上腺素受体 (α1AR) 激活 - 涉及激酶的蛋白质 C (PKC)-锚定蛋白 AKAP150–TRPV4 通道信号传导；和 (2) 扩张器纳米结构域 由腔内压力激活，包含以 Caveolin-1 为支架的 Piezo1–TRPV4–Ca2+- 此外，我们还发现了缩窄子α1AR-TRPV4 通道。 高血压和高血压小鼠模型的动脉信号传导增强 患者，而扩张器 TRPV4-BK 通道信号传导减少。 信号元件之间的沟通不畅破坏了收缩器和收缩器之间的平衡 在目标 1 中，扩张器信号纳米域会导致血压升高。 我们将使用平滑肌特异性基因敲除小鼠、蛋白质共定位、膜片钳 电生理学和血压无线电遥测以确定分子机制 导致收缩子 α1AR:AKAP150:PKC:TRPV4 纳米结构域过度激活 在目标 2 中，我们将确定导致血压降低的分子机制。 Caveolin-1 支架的扩张器 Piezo1:TRPV4:BK 通道纳米结构域的活性 每个目标的研究的临床相关性将通过以下实验确定。 总的来说，拟议的研究来自非高血压和高血压个体的动脉。 将建立新型 SMC Ca2+ 信号传导纳米域来控制血压并识别 高血压中这些纳米结构域的特定损伤。