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）是一种肺动脉血管疾病，其死亡率很高，最高为TO。 诊断三年后45％。 长期以来一直是PAH发展的主要贡献者。 肺动脉质量 - 一氧化氮（NO）中主要血管扩张分子的释放降低； 潜在的病理机制仍然未知。 调节肺血管中内皮NO合酶（ENOS）活性是破译的必要条件 在此应用中引起内皮功能障碍的机制。 通过内皮TRPV4（瞬态受体电势4）通道的涌入事件 - TRPV4 Sparklets - 调节eNOS活性，在肺动脉内皮中没有释放 揭示TRPV4是肺部内皮中的主要Ca2+涌入途径，可导致血管舒张 ENOS激活。 来自不同生理刺激的信息，包含三磷酸腺苷（ATP），流量/剪应力，以及 平滑肌1肾上腺素能通过不同的信号传导机制 血管舒张和没有释放在慢性缺氧引起的PAH中受损。 TRPV4 CA2+信号传导是在PAP中丧失了pap中的hothothothialialialialialialialialialialialiasialials。 确定原始治疗TRPV4通道作为关键控制机制的作用，该机制整合了信息 不同的生理刺激通过不同的信号通路引起小鼠和人的血管造成的刺激 肺动脉。 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 对生理刺激的反应。 通过内皮鸟类环酶 - 蛋白酶激酶G（PKG）机制，我们将描绘出来。 PAH小鼠模型中的TRPV4-ENOS功能障碍 上调导致PAH中TRPV4-Enos轴的功能受损。 新发现，包括局部CA2+信号调节eNOS的第一个证据及其在 PAH。研究的结果将提供急需的新型治疗靶标的忘记治疗PAH。

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