A novel TRPV4-eNOS signaling pathway in pulmonary endothelium

肺内皮细胞中新型 TRPV4-eNOS 信号通路

• 批准号: 9544363
• 负责人: Swapnil K. Sonkusare
• 金额: $ 40.27万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-02 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9544363
• 关键词: Adenosine Triphosphate; Adrenergic Agents; Adrenergic Receptor; Arteries; Biological Assay; Biosensor; Caliber; Cells; Chronic; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Data; Development; Diagnosis; Disease; Endothelial Cells; Endothelium; Event; Feedback; Functional disorder; Guanylate Cyclase; Human; Hypoxia; Image; Impairment; Ligation; Lung; Mechanics; Mediating; Mediator of activation protein; Mus; NOS3 gene; Nitric Oxide; Nitric Oxide Synthase; Pathologic; Pathway interactions; Pharmacology; Physiological; Progressive Disease; Pulmonary Circulation; Pulmonary Vascular Resistance; Pulmonary artery structure; Purinergic P2 Receptors; Receptor Signaling; Regulation; Resistance; Role; Signal Pathway; Signal Transduction; Smooth Muscle; Stimulus; Testing; Up-Regulation; Vanilloid; Vasodilation; Vasodilator Agents; constriction; design; endothelial dysfunction; improved; inhibitor/antagonist; mortality; mouse model; new therapeutic target; novel; novel therapeutics; patch clamp; pressure; pulmonary arterial hypertension; receptor; response; shear stress; tool; vasoconstriction

Abstract Pulmonary arterial hypertension (PAH) is a disease of pulmonary vasculature with a high mortality rate of up to 45% three years after diagnosis. PAH is often associated with the loss of endothelial vasodilation, which has long been thought to be a major contributor to development of PAH. Endothelial dysfunction in PAH results in reduced release of the predominant vasodilatory molecule in pulmonary vasculature– nitric oxide (NO); but the underlying pathological mechanisms remain unknown. A detailed mechanistic understanding of the pathways that regulate endothelial NO synthase (eNOS) activity in pulmonary vasculature is necessary for deciphering the mechanisms that cause endothelial dysfunction in PAH. In this application we focus on local, unitary Ca2+ influx events through endothelial TRPV4 (transient receptor potential vanilloid 4) channels–TRPV4 sparklets– that regulate eNOS activity and NO release in the native pulmonary artery endothelium. Our preliminary results reveal that TRPV4 is a major Ca2+ influx pathway in pulmonary endothelium that causes vasodilation through eNOS activation. Moreover, endothelial TRPV4 channels serve as a control mechanism that integrates information from different physiological stimuli, including adenosine triphosphate (ATP), flow/shear stress, and smooth muscle 1 adrenergic stimulation, via distinct signaling mechanisms. The TRPV4-induced vasodilations and NO release are impaired in chronic hypoxia-induced PAH. We hypothesize that impaired TRPV4 Ca2+ signaling is responsible for the loss of endothelial vasodilations in PAH. In Specific Aim 1, we will determine the role of endothelial TRPV4 channels as a key control mechanism that integrates information from different physiological stimuli via distinct signaling pathways to cause vasodilations in mouse and human pulmonary arteries. We will also confirm the physiological roles of endothelial TRPV4 channels in endothelium- specific TRPV4-/- mice. In Specific Aim 2, we will define the TRPV4-eNOS linkage that causes vasodilation in response to physiological stimuli. We will also determine how NO itself regulates TRPV4 channel activity through endothelial guanylyl cyclase-protein kinase G (PKG) mechanism. In Specific Aim 3, we will delineate the TRPV4-eNOS dysfunction in mouse models of PAH, and test the hypothesis that endothelial PKG upregulation is responsible for impaired function of TRPV4-eNOS axis in PAH. These studies will result in novel discoveries including first evidence of eNOS regulation by localized Ca2+ signals and its impairment in PAH. Results from these studies will provide much needed novel therapeutic targets for treating PAH.

抽象的 肺动脉高压（PAH）是一种肺血管疾病，死亡率高至 诊断三年后45％。 PAH通常与内皮血管舒张的损失有关 长期以来，人们被认为是PAH发展的主要贡献者。 PAH中的内皮功能障碍导致 肺脉管系统 - 一氧化氮（NO）中主要血管舒张分子的释放降低；但是 潜在的病理机制仍然未知。对路径的详细机械理解 调节肺脉管中的内皮NO合酶（ENOS）活性是破译的必要条件 PAH引起内皮功能障碍的机制。在此应用中，我们专注于本地的统一CA2+ 通过内皮TRPV4（瞬态接收器潜在的香草素4）通道– TRPV4 Sparklets - 调节eNOS活性，而在天然肺动脉森林中没有释放。我们的初步结果 揭示TRPV4是肺部内皮的主要CA2+影响途径，可通过 eNOS激活。此外，内皮TRPV4通道充当整合的控制机制 来自不同物理刺激的信息，包括三磷酸腺苷（ATP），流量/剪切应力和 平滑肌1肾上腺素刺激，通过不同的信号传导机制。 TRPV4引起的 在慢性缺氧诱导的PAH中，血管舒张和无释放受损。我们假设那障 TRPV4 CA2+信号传导负责PAH中内皮血管舒张的损失。在特定目标1中，我们将 确定内皮TRPV4通道作为关键控制机制的作用，该机制整合了信息 通过不同的信号传导途径不同的物理刺激，引起小鼠和人的血管舒张 肺动脉。我们还将确认内皮中内皮TRPV4通道的身体作用 特定的TRPV4 - / - 小鼠。在特定的目标2中，我们将定义引起血管舒张的TRPV4-ENOS链接 对身体刺激的反应。我们还将确定没有本身如何调节TRPV4通道活动 通过内皮鸟叶烯基环酶蛋白激酶G（PKG）机制。在特定目标3中，我们将描绘 PAH小鼠模型中的TRPV4-ENOS功能障碍，并测试了内皮PKG的假设 上调导致PAH中TRPV4-ENOS轴的功能受损。这些研究将导致 新发现，包括局部Ca2+信号调节eNOS调节的第一个证据及其在 pah。这些研究的结果将为治疗PAH提供急需的新型治疗靶标。

项目成果

Nitric Oxide-Dependent Feedback Loop Regulates Transient Receptor Potential Vanilloid 4 (TRPV4) Channel Cooperativity and Endothelial Function in Small Pulmonary Arteries.

• DOI: 10.1161/jaha.117.007157
• 发表时间: 2017-12-23
• 期刊: Journal of the American Heart Association
• 影响因子: 5.4
• 作者: Marziano C;Hong K;Cope EL;Kotlikoff MI;Isakson BE;Sonkusare SK
• 通讯作者: Sonkusare SK