TRP channels in the regulation of vascular tone

TRP 通道在血管张力调节中的作用

• 批准号: 10654013
• 负责人: David X. Zhang
• 金额: $ 53.07万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-04 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10654013
• 关键词: Address; Adipose tissue; Aging; Animal Disease Models; Animal Model; Arachidonic Acids; Arteries; Atherosclerosis; Binding; Binding Sites; Blood Vessels; Cause of Death; Cell Separation; Cells; Compensation; Coronary; Coronary Arteriosclerosis; Cyclic AMP-Dependent Protein Kinases; Disease; Electrophysiology (science); Endothelial Cells; Endothelium; Enzymes; Equilibrium; Functional disorder; Funding; Gene Expression; Genetic Polymorphism; Goals; Health; Homeostasis; Homologous Gene; Human; Hydrogen Peroxide; Image; Impairment; Inflammation; Ion Channel; Knock-out; Ligands; Mass Spectrum Analysis; Mediating; Mediator; Metabolism; Microcirculation; Microvascular Dysfunction; Mitochondria; Modeling; Molecular; Mutagenesis; Myocardial Ischemia; NADPH Oxidase; Nitric Oxide; Outcome; Oxidation-Reduction; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patient-Focused Outcomes; Patients; Phospholipase A2; Phosphorylation; Play; Process; Production; Protein Isoforms; Proteins; Reactive Oxygen Species; Regulation; Role; Signal Transduction; Small Interfering RNA; Structural Models; Structure; TRP channel; Testing; Transfection; Vanilloid; Variant; Vascular Diseases; Vasodilation; Vasodilator Agents; Woman; acute stress; analog; arteriole; design; eicosanoid metabolism; endothelial dysfunction; gain of function; human disease; human model; human tissue; improved; in vivo; inhibitor; innovation; insight; lipophilicity; men; mimetics; molecular modeling; novel; novel strategies; outcome prediction; patch clamp; pharmacologic; receptor; release factor; structural biology; synergism; targeted treatment; transcriptome sequencing; vasomotion

Vascular homeostasis is critically dependent upon vasodilator factors released from the endothelium. The most prominent of these factors is nitric oxide (NO), which is the main barometer of endothelial function and becomes impaired in a broad range of diseases including coronary artery disease (CAD). In the human coronary and adipose microcirculation, we have demonstrated a novel process where loss of NO-dependent flow-mediated dilation (FMD) in subjects with CAD is compensated by the production of hydrogen peroxide (H2O2) from endothelial mitochondria and subsequent H2O2-dependent dilation. Although both are vasodilators, H2O2, in opposition to NO, generally promotes cell activation, inflammation, and atherosclerosis, and thus understanding mechanisms responsible for this transition from NO to H2O2 may be key to developing novel strategies to improve endothelial function in patients with CAD. The overall goal of this project is to elucidate the signaling mechanisms that regulate the vasodilator switch from NO to H2O2 during CAD. Building on findings from the last cycle, this proposal is designed to determine intracellular pathways responsible for a previously unappreciated gain of function of endothelial transient receptor potential vanilloid 4 (TRPV4) channels and its contribution to vasodilator switch in CAD. We will test the central hypothesis that a synergy of shear-sensitive phospholipase A2-derived arachidonic acid and NADPH oxidase signaling promotes TRPV4 activation and subsequent H2O2-dependent dilation while cross-inhibiting NO-dependent dilation in CAD arterioles. Further, NADPH oxidases as novel aging- and CAD-associated upstream regulators play a critical role in initiating the switch. This application brings together expertise in vasomotion regulation, human microcirculation, and ion channel structural biology to identify novel molecular mechanisms and interactions that regulate vasodilator switch during CAD. Specific Aims: (1) we will determine the molecular mechanism of TRPV4 activation and arteriolar dilation by flow; and (2) we will determine how NADPH oxidases regulate TRPV4 activation and conversion from NO to H2O2 as mediator of FMD in CAD arterioles. Studies will be conducted on freshly isolated human arterioles and endothelial cells as well as in vivo animal models, using a multifaceted approach incorporating isolated vessel reactivity, Ca2+ imaging, patch-clamping electrophysiology, mass spectrometry, RNA-Seq, mutagenesis, and ion channel molecular modeling. Significance: our proposed studies will provide insight into fundamental mechanisms regulating human microvascular function in health and disease and potentially impact our approach to coronary microvascular dysfunction associated with CAD and a variety of other vascular pathologies.

血管稳态很大程度上取决于内皮释放的血管舒张因子。最 这些因素中最突出的是一氧化氮（NO），它是内皮功能的主要晴雨表，并成为 包括冠状动脉疾病（CAD）在内的多种疾病都会受到损害。在人体冠状动脉和 脂肪微循环，我们已经证明了一种新的过程，其中NO依赖的血流介导的损失 患有 CAD 的受试者的扩张 (FMD) 通过产生过氧化氢 (H2O2) 来补偿 内皮线粒体和随后的 H2O2 依赖性扩张。虽然两者都是血管扩张剂，但 H2O2 对抗 NO，通常会促进细胞活化、炎症和动脉粥样硬化，因此了解 负责从 NO 到 H2O2 转变的机制可能是开发新策略来改善的关键 CAD 患者的内皮功能。该项目的总体目标是阐明信号机制 在 CAD 期间调节血管舒张剂从 NO 切换为 H2O2。基于上一周期的调查结果，本次 该提案旨在确定细胞内途径，该途径导致以前未被认识到的增益 内皮瞬时受体电位香草酸 4 (TRPV4) 通道的功能及其对血管舒张的贡献 在 CAD 中切换。我们将测试核心假设，即剪切敏感磷脂酶 A2 衍生的协同作用 花生四烯酸和 NADPH 氧化酶信号传导促进 TRPV4 激活和随后的 H2O2 依赖性 扩张，同时交叉抑制 CAD 小动脉中 NO 依赖性扩张。此外，NADPH 氧化酶作为新型衰老- 与 CAD 相关的上游监管机构在启动转换中发挥着关键作用。这个应用程序带来了 结合血管舒缩调节、人体微循环和离子通道结构生物学方面的专业知识来确定 CAD 期间调节血管舒张开关的新分子机制和相互作用。具体目标：（1）我们 将确定TRPV4激活和血流扩张小动脉的分子机制； (2) 我们将 确定 NADPH 氧化酶如何调节 TRPV4 激活以及从 NO 到 H2O2 的转化作为介导 CAD 小动脉中的 FMD。研究将对新鲜分离的人小动脉和内皮细胞进行 以及体内动物模型，使用多方面的方法结合分离的血管反应性、Ca2+ 成像、膜片钳电生理学、质谱、RNA-Seq、诱变和离子通道 分子建模。意义：我们提出的研究将提供对基本机制的见解 调节健康和疾病中的人体微血管功能，并可能影响我们治疗冠状动脉的方法 与 CAD 和各种其他血管病变相关的微血管功能障碍。

项目成果

NADPH oxidase 4 contributes to TRPV4-mediated endothelium-dependent vasodilation in human arterioles by regulating protein phosphorylation of TRPV4 channels.

• DOI: 10.1007/s00395-022-00932-9
• 发表时间: 2022-04-25
• 期刊: Basic research in cardiology
• 影响因子: 9.5

Characterization of blood pressure and endothelial function in TRPV4-deficient mice with l-NAME- and angiotensin II-induced hypertension.

• DOI: 10.1002/phy2.199
• 发表时间: 2014-01-01
• 期刊: PHYSIOLOGICAL REPORTS
• 影响因子: 2.5
• 作者: Nishijima, Yoshinori;Zheng, Xiaodong;Lund, Hayley;Suzuki, Makoto;Mattson, David L;Zhang, David X
• 通讯作者: Zhang, David X

H2O2 is the transferrable factor mediating flow-induced dilation in human coronary arterioles.

• DOI: 10.1161/circresaha.110.237636
• 发表时间: 2011-03-04
• 期刊: Circulation research
• 影响因子: 20.1
• 作者: Liu Y;Bubolz AH;Mendoza S;Zhang DX;Gutterman DD
• 通讯作者: Gutterman DD

Rap1b in smooth muscle and endothelium is required for maintenance of vascular tone and normal blood pressure.

• DOI: 10.1161/atvbaha.114.303678
• 发表时间: 2014-07
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Lakshmikanthan S;Zieba BJ;Ge ZD;Momotani K;Zheng X;Lund H;Artamonov MV;Maas JE;Szabo A;Zhang DX;Auchampach JA;Mattson DL;Somlyo AV;Chrzanowska-Wodnicka M
• 通讯作者: Chrzanowska-Wodnicka M

Arachidonic acid-induced dilation in human coronary arterioles: convergence of signaling mechanisms on endothelial TRPV4-mediated Ca2+ entry.

• DOI: 10.1161/jaha.113.000080
• 发表时间: 2013-04-25
• 期刊: Journal of the American Heart Association
• 影响因子: 5.4
• 作者: Zheng X;Zinkevich NS;Gebremedhin D;Gauthier KM;Nishijima Y;Fang J;Wilcox DA;Campbell WB;Gutterman DD;Zhang DX
• 通讯作者: Zhang DX