Nitric Oxide in Pulmonary Hypertension

一氧化氮与肺动脉高压

• 批准号: 10330030
• 负责人: Serpil C. Erzurum
• 金额: $ 77.2万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10330030
• 关键词: ARG2 gene; Affect; Altitude; Arginine; Award; Big Data; Bioenergetics; Biological Availability; Blood; Cardiac; Cells; Cellular Metabolic Process; Cessation of life; Citric Acid Cycle; Data; Dependence; Development; Disease; Electron Transport; Endothelium; Erythropoietin; Functional disorder; Glucose; Glutamates; Goals; Grant; Harvest; Hemoglobin; Human; Hypoxia; Hypoxia Inducible Factor; Knock-out; Knockout Mice; Knowledge; Left; Link; Lung; Measures; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Methods; Mitochondria; Multiomic Data; Mus; NOS3 gene; Nature; Nitric Oxide; Nitric Oxide Synthase; Ornithine; Oxygen Consumption; Pathologic; Pathway Analysis; Pathway interactions; Patients; Persons; Phenotype; Phosphorylation; Production; Pulmonary Hypertension; RNA; Resistance; Resource Sharing; Respiration; Right Ventricular Dysfunction; System; Systolic Pressure; Testing; Transplantation; Tricarboxylic Acids; Vasodilator Agents; Ventricular; Wild Type Mouse; Woman; Work; aerobic glycolysis; arginase; differential expression; experimental study; fluorodeoxyglucose positron emission tomography; glucose uptake; hypertension control; hypoxia-induced pulmonary hypertension; improved; in vivo; lung pressure; metabolomics; method development; multiple omics; nitric oxide synthase II; oxidation; protein metabolite; pulmonary arterial hypertension; pulmonary artery endothelial cell; pulmonary vascular cells; therapeutic target; trait; transcriptomics; treatment strategy; uptake

ABSTRACT This is a competitive renewal of our R37 MERIT (2009-2019), which aims to define mechanisms of Pulmonary Arterial Hypertension (PAH). Our work led to new knowledge, shared resources and methods, including a critical advance in harvest and culture of human pulmonary artery endothelial cells (PAEC). We identified loss of nitric oxide (NO) production, and mechanisms: (i) phosphorylation inactivation of endothelial NO synthase (eNOS), and (ii) decreased eNOS substrate arginine (arg) availability related to increased mitochondrial arginase 2 (ARG2). We discovered that high-altitude natives, who avoid high-altitude hypoxic pulmonary hypertension (HAPH), have adaptations that increase arg, NO, and decrease ARG2. In parallel, we found that PAH PAEC have less mitochondrial respiration than control cells, and a shift to aerobic glycolysis. In vivo, lung and cardiac glucose uptake in PAH patients measured by 18F-fluorodeoxyglucose-positron emission tomography (FDG-PET) was higher than controls. In preliminary data, patients with the highest FDG uptake have the lowest arg levels and most severe disease, suggesting that arg metabolic fate impacts PAH beyond just the loss of vasodilator NO. We show that mitochondrial arg metabolism is interconnected to bioenergetics, including fuel dependency of tricarboxylic acid cycle (TCA) and mitochondrial respiration. In murine studies, Arg2 knockout (Arg2KO) have greater capacity for mitochondrial respiration and less cardiac glucose uptake than wildtype (WT), and lack the hypoxia-induced increases of pulmonary pressures and erythropoietin (Epo) found in WT. Thus, we hypothesize that mitochondrial arg metabolism via ARG2 is interconnected to abnormalities of TCA cycle and bioenergetics, and promotes pathologic proliferation of PAEC, development of PAH and right ventricular (RV) dysfunction. Aim 1 identifies disease metabolic mechanisms and pathways that participate in pathologic endothelial functions and PAH pathophysiology. To find pathways associated with PAH, we perform differential expression (RNA, protein, metabolites) and integrative network analyses of PAH and control PAEC, then measure cell bioenergetics and functions after blocking pathways enriched in PAH (Aim1A). PAH patients, dichotomized into low or high cardiac FDG uptake groups, are compared with controls using metabolomic and transcriptomic analyses to uncover pathways in vivo, then followed longitudinally to determine if death, transplant, and/or worsening RV systolic pressure (RVSP) is greater in the high uptake group (Aim1B). Aim 2 determines if decreasing mitochondrial arg metabolism is protective against hypoxia-associated PH. Metabolomic profiles of high-altitude Amhara, who have high NO, low arginase, and resist HAPH, are compared to their low-altitude counterparts and PAH patients, to identify protective pathways in relation to quantitative traits, such as Epo and RVSP (Aim 2A). In mechanistic studies, we determine if Arg2KO have bioenergetic changes as compared to WT, and if Arg2KO are protected from hypoxia-induced elevations in Epo and pulmonary pressures (Aim 2B). These studies will provide new metabolic understanding of PAH, and offer new potential therapeutic targets.

抽象的 这是我们的 R37 MERIT (2009-2019) 的竞争性更新，旨在定义肺功能的机制 动脉高血压（PAH）。我们的工作带来了新知识、共享资源和方法，包括关键的 人肺动脉内皮细胞（PAEC）的收获和培养取得进展。我们发现硝酸盐损失 氧化物（NO）的产生和机制：（i）内皮NO合酶（eNOS）的磷酸化失活， (ii) eNOS 底物精氨酸 (arg) 可用性降低与线粒体精氨酸酶 2 增加相关 （ARG2）。我们发现，高原人，避免高原缺氧性肺动脉高压 (HAPH)，具有增加 arg、NO 并减少 ARG2 的适应性。与此同时，我们发现 PAH PAEC 与对照细胞相比，线粒体呼吸较少，并且转向有氧糖酵解。体内、肺和心脏 通过 18F-氟脱氧葡萄糖-正电子发射断层扫描 (FDG-PET) 测量 PAH 患者的葡萄糖摄取 高于对照。初步数据显示，FDG 摄取量最高的患者的 arg 水平最低 和最严重的疾病，表明 arg 代谢命运对 PAH 的影响不仅仅是血管扩张剂的损失 不。我们表明线粒体精氨酸代谢与生物能学相互关联，包括燃料依赖性 三羧酸循环（TCA）和线粒体呼吸。在小鼠研究中，Arg2 敲除 (Arg2KO) 与野生型（WT）相比，线粒体呼吸能力更强，心脏葡萄糖摄取更少，并且缺乏 WT 中发现缺氧引起的肺压和促红细胞生成素 (Epo) 增加。因此，我们假设 通过 ARG2 进行的线粒体精氨酸代谢与 TCA 循环和生物能异常相关， 并促进 PAEC 的病理性增殖、PAH 的发生和右心室 (RV) 功能障碍。目的 1 确定参与病理内皮功能的疾病代谢机制和途径 PAH 病理生理学。为了找到与 PAH 相关的途径，我们进行差异表达（RNA、蛋白质、 代谢物）以及 PAH 和对照 PAEC 的综合网络分析，然后测量细胞生物能和 阻断富含 PAH (Aim1A) 的途径后发挥作用。 PAH 患者分为低心脏病和高心脏病 使用代谢组学和转录组学分析将 FDG 摄取组与对照组进行比较，以揭示 体内途径，然后纵向追踪以确定死亡、移植和/或 RV 收缩压恶化是否 高摄取组 (Aim1B) 的压力 (RVSP) 更大。目标 2 确定是否减少线粒体 arg 新陈代谢对缺氧相关的PH具有保护作用。高海拔阿姆哈拉人的代谢组学概况 高NO、低精氨酸酶和抵抗HAPH的患者与低海拔人群和PAH患者进行比较， 确定与数量性状相关的保护途径，例如 Epo 和 RVSP（目标 2A）。在机械论上 研究中，我们确定 Arg2KO 与 WT 相比是否具有生物能变化，以及 Arg2KO 是否受到保护 缺氧引起的 Epo 和肺压升高（目标 2B）。这些研究将提供新的 对 PAH 的代谢了解，并提供新的潜在治疗靶点。