Aldosterone impairs endothelin B-dependent synthesis of nitric oxide to promote p

醛固酮损害内皮素 B 依赖性一氧化氮合成，促进 p

• 批准号: 8442173
• 负责人: Bradley Maron
• 金额: $ 13.62万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8442173
• 关键词: Adrenal Glands; Affect; Aldosterone; Angiotensins; Basic Science; Bioavailable; Biochemistry; Biology; Blood Circulation; Blood Pressure; Blood Vessels; Cardiopulmonary; Cardiovascular Diseases; Cardiovascular system; Cellular biology; Cessation of life; Clinical; Clinical Research; Congestive Heart Failure; Coupling; Data; Development; Disease; Distal; Endothelin; Endothelin B Receptor; Endothelin-1; Endothelium; Ensure; Environment; Faculty; Fellowship; Functional disorder; Future; Generations; Heart Diseases; Hematological Disease; Hormones; Hospitals; Human; Hypoxia; In Vitro; Inflammatory; Injury; K-Series Research Career Programs; LIM Domain; Laboratories; Laboratory Research; Left; Link; Lung; Lung diseases; Mediating; Medicine; Mentors; Mentorship; Mineralocorticoid Receptor; Modeling; Modification; Molecular; Monocrotaline; Morbidity - disease rate; NADPH Oxidase; Nitric Oxide; Oxidation-Reduction; Patients; Physicians; Plasma; Principal Investigator; Production; Proteins; Proteomics; Pulmonary Circulation; Pulmonary Hypertension; Pulmonary Vascular Resistance; Pulmonary artery structure; Quality of life; Rattus; Reactive Oxygen Species; Receptor Signaling; Relaxation; Renin; Research; Resources; Scientist; Side; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Smooth Muscle Myocytes; Spironolactone; Sulfenic Acids; Sulfhydryl Compounds; Testing; Therapeutic Effect; Training Programs; United States National Institutes of Health; Vascular Diseases; Vascular Endothelium-Dependent Relaxation; Vasodilation; Woman; arteriole; career; career development; design; disulfide bond; experience; hemodynamics; human NOS3 protein; improved; in vivo; innovation; liquid chromatography mass spectrometry; medical schools; mortality; novel; oxidant stress; oxidation; patient population; premature; pressure; professor; programs; protein function; public health relevance; pulmonary arterial hypertension; pulmonary artery endothelial cell; receptor; success; transmission process

DESCRIPTION (provided by applicant): The NIH Mentored Clinical Scientist Research Career Development Award proposal describes a 5-year training program for career development in academic cardiovascular medicine. The principal investigator (PI) completed a postdoctoral basic science research fellowship (2005-2008) and in July 2011 joined as Faculty in the Division of Cardiovascular Medicine at Brigham and Women's Hospital/Harvard Medical School (BWH/HMS). The PI continues an intensive research program (2010- ) designed to promote a career as an independent scientist in cardiovascular diseases. The proposed research plan affords the PI rigorous experience in cell biology, liquid chromatography-mass spectrometry (LC-MS), and cardiopulmonary hemodynamic assessment in vivo. Drs. Joseph Loscalzo and Jane Leopold will mentor the PI's scientific development during this period. Dr. Loscalzo is an internationally recognized leader in the fields of redox biology and pulmonary arterial hypertension (PAH), who has vast experience in mentoring successfully young physician-scientists. Dr. Leopold is an Associate Professor of Medicine (HMS) and an expert in the field of aldosterone (ALDO)-mediated vascular dysfunction, with a strong track record in mentorship. This proposal focuses on redox biochemistry to investigate the contribution of ALDO to the development of pulmonary vascular dysfunction in PAH; thus, co-mentorship for this proposal is uniquely suited for the PI. The Loscalzo/Leopold Cardiovascular Research Laboratory (BWH/HMS) is the site for this project and an excellent environment with the necessary resources to ensure the PI's success for achieving independence in academic medicine. PAH is a fatal disease characterized by increased pulmonary vascular endothelial reactive oxygen species (ROS) formation, decreased levels of bioavailable nitric oxide (NO.), and impaired pulmonary endothelium-dependent vasodilation. ROS-mediated modifications in the redox state of functionally essential protein cysteinyl thiols involved in NO. signaling may affect adversely vascular tone. In human pulmonary artery endothelial cells (HPAECs), endothelin- B (ETB) receptor Cys405 is functionally linked to activation of endothelial nitric oxide synthase by ETB to generate NO.. Here, we provide novel evidence to demonstrate that hyperALDO is present in PAH in vivo, which increases ROS levels in HPAECs to induce ETB disulfide bond/sulfenic acid formation and decrease ETB-dependent NO. synthesis. Thus, the central hypothesis of this proposal is that: In PAH, oxidative modification of Cys405 by hyperALDO- induced ROS formation acts as a molecular "switch" to disrupt ETB-dependent NO. generation and impair pulmonary vasodilation. The specific aims are: (1) investigate the functional effects of ALDO-induced ETB cysteinyl thiol oxidative modification(s) in vitro; (2a) determine the contribution of hyperALDO to impaired pulmonary vascular reactivity in PAH in vivo; and (2b) investigate the therapeutic effects of ALDO antagonism for PAH in vivo. We will use two different models of PAH in rats in vivo and LC-MS to identify ETB cysteinyl thiol oxidation product(s). These studies aim to identify novel treatment targets for PAH and other vascular diseases with similar pathobiology.

描述（由申请人提供）：NIH 指导临床科学家研究职业发展奖提案描述了学术心血管医学职业发展的 5 年培训计划。主要研究者 (PI) 完成了博士后基础科学研究奖学金（2005-2008 年），并于 2011 年 7 月加入布莱根妇女医院/哈佛医学院 (BWH/HMS) 心血管医学科担任教员。 PI 继续开展一项深入研究计划（2010 年至今），旨在促进心血管疾病领域独立科学家的职业生涯。拟议的研究计划为 PI 提供了细胞生物学、液相色谱-质谱 (LC-MS) 和体内心肺血流动力学评估方面的严格经验。博士。 Joseph Loscalzo 和 Jane Leopold 将指导 PI 在此期间的科学发展。 Loscalzo 博士是氧化还原生物学和肺动脉高压 (PAH) 领域国际公认的领导者，在成功指导年轻医师科学家方面拥有丰富的经验。 Leopold 博士是医学副教授 (HMS) 和醛固酮 (ALDO) 介导的血管功能障碍领域的专家，在指导方面拥有良好的记录。该提案重点关注氧化还原生物化学，研究 ALDO 对 PAH 肺血管功能障碍发展的贡献；因此，该提案的共同指导特别适合 PI。 Loscalzo/Leopold 心血管研究实验室 (BWH/HMS) 是该项目的所在地，拥有良好的环境和必要的资源，以确保 PI 成功实现学术医学的独立性。 PAH 是一种致命性疾病，其特征是肺血管内皮活性氧 (ROS) 形成增加、生物可利用一氧化氮 (NO.) 水平降低以及肺内皮依赖性血管舒张受损。 ROS 介导的参与 NO 的功能必需蛋白半胱氨酰硫醇氧化还原状态的修饰。信号传导可能会影响 不利的血管张力。在人肺动脉内皮细胞 (HPAEC) 中，内皮素 B (ETB) 受体 Cys405 在功能上与 ETB 激活内皮一氧化氮合酶产生 NO 相关。在此，我们提供新的证据来证明 hyperALDO 存在于体内 PAH 中，这会增加 HPAEC 中的 ROS 水平，从而诱导 ETB 二硫键/次磺酸的形成 并减少 ETB 依赖性 NO。合成。因此，该提议的中心假设是：在 PAH 中，hyperALDO 诱导的 ROS 形成对 Cys405 的氧化修饰充当破坏 ETB 依赖性 NO 的分子“开关”。产生并损害肺血管舒张。具体目的是：（1）体外研究ALDO诱导的ETB半胱氨酰硫醇氧化修饰的功能效应； (2a)确定hyperALDO对PAH体内肺血管反应性受损的贡献； (2b) 研究 ALDO 拮抗剂对 PAH 的体内治疗效果。我们将使用大鼠体内两种不同的 PAH 模型和 LC-MS 来鉴定 ETB 半胱氨酰硫醇氧化产物。这些研究旨在确定 PAH 和具有相似病理学的其他血管疾病的新治疗靶点。