Nitric Oxide in Pulmonary Hypertension

一氧化氮与肺动脉高压

• 批准号: 8112681
• 负责人: Serpil C. Erzurum
• 金额: $ 37.83万
• 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 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8112681
• 关键词: Abnormal Cell; Accounting; Altitude; Antioxidants; Apoptosis; Biochemical; Biological Models; Blood Circulation; Blood Vessels; Bone Marrow; Boxing; Caring; Cell Culture Techniques; Cell Line; Cell Respiration; Cell Survival; Cell model; Cells; Clinical; Clinical Research; Data; Disease; Endothelial Cells; Enzymes; Erythropoietin; Event; Figs - dietary; Fostering; Functional disorder; Glucose; Glycolysis; Goals; Grant; Gray unit of radiation dose; Growth; Hepatocyte Growth Factor; Human; Hypoxia; Hypoxia Inducible Factor; Immunodeficient Mouse; Implant; In Vitro; Knowledge; Link; Lung; Lung Transplantation; Malignant Neoplasms; Manganese Superoxide Dismutase; Measures; Metabolic; Metabolism; Methods; Mitochondria; Modeling; Molecular; Mus; Mutation; NOD/SCID mouse; Nature; Nitric Oxide; Nitric Oxide Synthase; Normal Cell; Outcome; Pathologic; Pathway interactions; Patient Care; Patients; Phenotype; Phosphorylation; Phosphorylation Inhibition; Physiological; Plasma; Population; Positron-Emission Tomography; Primary Cell Cultures; Production; Proteins; Pulmonary Hypertension; Pulmonary artery structure; Regulation; Research; Resistance; Respiration; STAT3 gene; Signal Transduction; Stat3 protein; Testing; Text; Threonine; Tibet; Time; Transducers; Vascular Diseases; Vascular Endothelial Cell; Vasodilator Agents; Work; Xenograft Model; Xenograft procedure; angiogenesis; autocrine; base; caveolin 1; glucose analog; glucose uptake; hemodynamics; human NOS3 protein; hypertension control; hypoxia inducible factor 1; improved; in vivo; matrigel; multidisciplinary; paracrine; patient population; pressure; progenitor; programs; public health relevance; pulmonary arterial hypertension; pulmonary artery endothelial cell; response; translational study

DESCRIPTION (provided by applicant): Pulmonary arterial hypertension (PAH) is a fatal disease associated with increased pulmonary artery pressure (PAP) and abnormalities in blood vessel growth. The excessive growth of the vascular endothelial cells is the focus of our research. Four years ago, we established methods for culture of pulmonary artery endothelial cells (PAEC) from PAH and donor lungs. The PAEC in culture reflect abnormalities found in PAH lungs and patients in vivo, and made it possible to test our hypotheses in translational mechanistic studies. We found that PAH PAEC have enhanced proliferation & survival, which is dependent on activation of signal transducer and activator of transcription-3 (STAT3), a fundamental regulator of cell survival and angiogenesis. The PAH cells have impaired production of the vasodilator nitric oxide (NO) due to post- translational mechanisms that include phosphorylation/inhibition of endothelial NO synthase (eNOS). In association with low NO, the cells have reduced mitochondria numbers & respiration, which is accompanied by ~3-fold increase in glycolysis for energy production. The shift to anaerobic glycolytic metabolism is related to expression of the pro-survival and pro-angiogenic signal transducer, hypoxia-inducible factor -1a (HIF-1a). Physiologic effects classically ascribed to HIF-expression [increased lung glucose uptake by 18F- fluoro-deoxy-glucose analogue - positron emission tomography (FDG-PET), high-levels of plasma erythropoietin (Epo) and mobilization of bone marrow progenitors] are present in patients and quantitatively associated with PAP. The HIF-expression is mechanistically linked to the alterations in STAT3, NO and Mn superoxide dismutase (MnSOD). Preliminary data show that the PAH PAEC secrete the potent growth- stimulatory & bone marrow-mobilizing hepatocyte growth factor (HGF). Present at high-levels in patient plasma and PAH cell cultures, HGF signals through STAT3 and induces HIF-1a, and is itself induced by STAT3 and increased by hypoxia, which endows the cells with autocrine and paracrine proliferative and pro-angiogenic effects. Here, we hypothesize that PAH endothelial cells have decreased NO production by eNOS and activation of survival pathways STAT3 and HIF-1a. These biochemical mechanisms account for the proliferative, glycolytic, and strongly pro-angiogenic phenotype of the cells. We identify mechanisms of reduced eNOS activity in aim 1, and focus on HIF-expression and mechanisms accounting for its expression in aim 2. In aim 3, we test if HIF-expression is mechanistically important in PAH pathophysiology using the primary cell culture model, a xenograft model of PAH cells in immunodeficient mice and a longitudinal clinical study, in which we test whether experimental outcomes of HIF-effects are associated with clinical outcomes in patients over time. Overall our goals are to define the pathophysiology of the abnormal vascular growth in PAH, and in so doing, apply the knowledge to improve the care of patients. PUBLIC HEALTH RELEVANCE: Pulmonary arterial hypertension (PAH) is a fatal disease associated with increased pulmonary artery pressure and abnormalities in blood vessel growth. The excessive growth of the endothelial cells that line the blood vessels in the lungs of patients afflicted with PAH is the focus of our research. Our studies provide evidence for biochemical and metabolic abnormalities in vascular endothelial cells in PAH, and identify the causal molecular mechanisms. We have found that the PAH endothelial cells have lower than normal production of the vasoregulatory gaseous molecule nitric oxide (NO), and use more than 3-fold greater amounts of glucose for energy production than normal cells. On the basis of our findings, we propose basic biochemical studies, and also test whether measures of glucose uptake in the lung using intravenously injected glucose analogues and positron emission tomography (FDG-PET) are useful to assess lung vascular disease in patients over time. Our goals in this competitive renewal are to define the pathophysiology of the abnormal vascular growth, and in so doing, apply the knowledge to improve the care of patients.

描述（由申请人提供）：肺动脉高压（PAH）是一种致命疾病，与血管生长的肺动脉压力增加（PAP）和异常有关。血管内皮细胞的过度生长是我们研究的重点。四年前，我们建立了PAH和供体肺的肺动脉内皮细胞（PAEC）培养方法。培养物中的PAEC反映了PAH肺和体内患者的异常，并使我们可以在翻译机械研究中检验我们的假设。我们发现PAH PAEC增强了增殖和生存，这取决于信号传感器的激活和转录3的激活因子（Stat3）（STAT3），这是细胞存活和血管生成的基本调节剂。 PAH细胞由于转化后机制而导致的血管扩张剂一氧化氮（NO）的产生受损，包括磷酸化/抑制内皮NO合酶（ENOS）。与低NO相关，细胞的线粒体数量和呼吸减少，伴随着糖酵解的能量产生约3倍。向厌氧糖酵解代谢的转变与促生存和血管生成信号传感器的表达有关，低氧诱导因子-1A（HIF-1A）。经典地归因于HIF表达的生理效应[18f-氟脱氧 - 葡萄糖类似物的肺葡萄糖摄取增加 - 正电子发射断层扫描（FDG-PET），血浆促红细胞增生蛋白（EPO）的高水平和骨骨髓的动员在患者中，与PAP定量相关。 HIF表达与STAT3，NO和MN超氧化物歧化酶（MNSOD）的改变有关。初步数据表明，PAH PAEC分泌有效的生长刺激性和骨髓化肝细胞生长因子（HGF）。存在于患者血浆和PAH细胞培养物中的高级水平，通过STAT3进行HGF信号并诱导HIF-1A，并且本身是由STAT3诱导的，并由缺氧增加，这会赋予细胞自动分泌和旁分泌增殖和促促血管生成作用。在这里，我们假设PAH内皮细胞通过eNOS和生存途径STAT3和HIF-1A的激活降低了没有产生。这些生化机制解释了细胞的增殖，糖酵解和强烈的血管生成表型。我们确定AIM 1中eNOS活性降低的机制，并专注于HIF表达和在AIM 2中表达的机制。在AIM 3中，我们使用原代细胞培养模型在PAH病理生理学中测试HIF表达在机械上很重要，免疫缺陷小鼠中PAH细胞的异种移植模型和一项纵向临床研究，其中我们测试了HIF效应的实验结果是否与患者的临床结局有关。总体而言，我们的目标是定义PAH异常血管生长的病理生理学，并在这样做的过程中使用知识来改善患者的护理。 公共卫生相关性：肺动脉高压（PAH）是一种致命疾病，与肺动脉压力增加和血管生长异常有关。在患有PAH的患者肺中排列血管的内皮细胞的过度生长是我们研究的重点。我们的研究为PAH血管内皮细胞中的生化和代谢异常提供了证据，并鉴定了因果分子机制。我们发现，PAH内皮细胞的血管调节气态分子一氧化氮（NO）的产生低于正常产生，并且使用比正常细胞使用的葡萄糖大量超过3倍以上。根据我们的发现，我们提出了基本的生物化学研究，还可以使用静脉注射的葡萄糖类似物和正电子发射断层扫描（FDG-PET）测试肺中葡萄糖摄取的量度是否有用，可用于评估患者的肺血管疾病。我们在这种竞争性更新中的目标是定义异常血管生长的病理生理，因此，将知识应用于改善患者的护理。