Reactive Oxygen Species in Coronary Collateral Growth

冠状动脉侧枝生长中的活性氧

• 批准号: 7754159
• 负责人: WILLIAM M CHILIAN
• 金额: $ 45.02万
• 依托单位: NORTHEAST OHIO MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-22 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7754159
• 关键词: Abbreviations; Adenoviruses; Animal Model; Animals; Anterior Descending Coronary Artery; Antioxidants; Arginine; Binding; Biology; Blood Vessels; Bypass; Canis familiaris; Cardiac; Cardiac Myocytes; Cardiac Myosins; Cardiovascular system; Cause of Death; Cells; Chronic; Collateral Circulation; Coronary; Data; Dependency; Development; Disease; Dominant-Negative Mutation; Dyslipidemias; Electroporation; Endothelial Cells; Endothelium; Environment; Event; Foundations; Glossary; Goals; Growth; Growth Factor; Hand; Healthcare Systems; Heart; Hydrogen Peroxide; Hyperglycemia; Hyperlipidemia; Hypertension; Impairment; Incidence; Infarction; Insulin Resistance; Investigation; Ischemia; Laboratories; Left; Link; Location; MAP Kinase Gene; MAPK14 gene; Measurement; Mediator of activation protein; Metabolic syndrome; Mitogen-Activated Protein Kinases; Modeling; Morbidity - disease rate; Muscle Cells; Mutate; Myocardial; Myocardial Ischemia; Myocardium; Myosin Heavy Chains; NF-E2-related factor 2; Obesity; Oxidation-Reduction; Oxidative Stress; Pathology; Patients; Plasmid Cloning Vector; Plasmids; Problem Solving; Production; Protocols documentation; Rattus; Reactive Oxygen Species; Reperfusion Therapy; Reporting; Response Elements; Risk Factors; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Specificity; Sudden Death; Sulfhydryl Compounds; Techniques; Testing; United States; Vascular Diseases; Viral; base; cadherin 5; cell type; cost; dihydroethidium; enhanced green fluorescent protein; falls; gene therapy; human MAPK14 protein; in vivo; insight; mitogen-activated protein kinase p38; mortality; mutant; nuclear factor-erythroid 2; outcome forecast; oxidation; prevent; promoter; protein activation; protein expression; public health relevance; research study; response; vector

DESCRIPTION (provided by applicant): Results from our and other laboratories demonstrate that a critical amount of ROS and a redox state within a certain boundary is critical for coronary collateral growth; however, oxidative stress, i.e., a shift in the redox state to a more oxidative environment, impairs coronary collateral growth. Previous investigations fall short of ascertaining the cell type (or types), in which alterations of redox state matter, and where redox signaling is critical. The overarching goal of this proposal is to determine the cell type or types in the heart responsible for redox signaling in the growth of the coronary collateral circulation in response to repetitive ischemia. A corollary to this aim is that we will also determine the cell type or types in which oxidative stress confers negative influences on coronary collateral growth. To solve these problems we propose the following specific aims: Aim 1. Determine in which cell type (or types) does oxidative stress corrupt coronary collateral growth. We will induce oxidative stress in the coronary endothelium, smooth muscle cells and cardiac myocytes using the cell-specific promoters VE-Cadherin, SM22, and cardiac myosin heavy chain (CMHC), respectively. Cells will be transfected with a plasmid (using in vivo electroporation) or transduced with an adenovirus expressing an iNOS mutant (E371A) that does not bind arginine, and therefore, only produces O2. Aim 2. Determine in which cell type (or types) the redox sensitive p38 MAPK is critical for coronary collateral growth. We will transfect or transduce coronary endothelium, smooth muscle cells and cardiac myocytes using cell-specific promoters described for Aim 1 using a vector expressing a dominant/negative p38 MAPK (DNp38). After determining the particular cell type in the heart most sensitive to the effects of oxidative stress and redox signaling in coronary collateral growth, we will extend our findings to an animal model of vascular pathology (the JCR rat: a model of the metabolic syndrome), which demonstrates poor coronary collateral growth. Specifically in the final two aims we will: Aim 3. Determine the cell type where reducing oxidative stress by over expressing Nrf2 in the JCR rat (a model of reduced coronary collateral growth and oxidative stress) will restore collateral growth. Aim 4. Determine the cell type where expression of a constitutively active p38 MAP kinase will restore collateral growth in the JCR rat. The proposed studies employ a multifaceted approach to solving cell-specific signaling in vivo by employing techniques to determine cell specific changes in protein expression, thiol oxidation and ROS production and ultimately linking these measurements to coronary collateral growth. These studies will provide insight into the cell-specific locations where ROS and redox signaling modulate coronary collateral growth. PUBLIC HEALTH RELEVANCE: Ischemic heart disease (IHD) continues to be one of the leading causes of death in the United States, and represents a major cost in the US healthcare system. The presence of a well developed collateral circulation in the heart has a tremendous impact on the morbidity and mortality of IHD. Patients with well developed collaterals show a lower incidence of sudden death, have smaller infarcts in the event of an occlusion, and demonstrate a better prognosis than patients with poor collaterals (low conductance or evidence of collateral growth). It is also worth noting that about 40% of patients with IHD demonstrate very little to no coronary collaterals. It is also recognized that many risk factors for IHD, e.g., hypertension, obesity, dyslipidemia, which also produce oxidative stress, have a negative influence on coronary collateral growth. However, the cell type where the corruption occurs, e.g., does oxidative stress corrupt the production of growth factors by cardiac myocytes, or does it corrupt endothelial responsiveness to growth factors, remains unknown. The goal of this proposal is to delineate the cell type in the heart where redox signaling and oxidative stress matter for coronary collateral growth. The ultimate goal of this project is to provide the foundation for more directed therapies to stimulate the growth of coronary collaterals in patients with IHD.

描述（由申请人提供）：我们和其他实验室的结果表明，临界量的 ROS 和一定范围内的氧化还原状态对于冠状动脉侧枝生长至关重要；然而，氧化应激，即氧化还原状态转变为氧化性更强的环境，会损害冠状动脉侧枝生长。以前的研究未能确定细胞类型（或多种细胞类型），其中氧化还原状态的改变很重要，以及氧化还原信号传导在哪些细胞中至关重要。该提案的总体目标是确定心脏中负责响应重复缺血的冠状动脉侧支循环生长中的氧化还原信号传导的细胞类型。这一目标的必然结果是，我们还将确定氧化应激对冠状动脉侧枝生长产生负面影响的细胞类型。为了解决这些问题，我们提出以下具体目标： 目标 1. 确定哪种细胞类型（或多种细胞）氧化应激会破坏冠状动脉侧枝生长。我们将分别使用细胞特异性启动子 VE-钙粘蛋白、SM22 和心肌肌球蛋白重链 (CMHC) 诱导冠状动脉内皮、平滑肌细胞和心肌细胞中的氧化应激。细胞将用质粒转染（使用体内电穿孔）或用表达 iNOS 突变体 (E371A) 的腺病毒转导，该突变体不结合精氨酸，因此仅产生 O2。目标 2. 确定哪种（或多种）细胞类型中氧化还原敏感的 p38 MAPK 对冠状动脉侧枝生长至关重要。我们将使用目标 1 中描述的细胞特异性启动子，使用表达显性/阴性 p38 MAPK (DNp38) 的载体转染或转导冠状动脉内皮、平滑肌细胞和心肌细胞。在确定了心脏中对冠状动脉侧枝生长中氧化应激和氧化还原信号的影响最敏感的特定细胞类型后，我们将把我们的发现扩展到血管病理学动物模型（JCR大鼠：代谢综合征模型），这表明冠状动脉侧枝生长不良。具体来说，在最后两个目标中，我们将： 目标 3. 确定通过在 JCR 大鼠（冠状动脉侧枝生长和氧化应激减少的模型）中过度表达 Nrf2 来减少氧化应激将恢复侧枝生长的细胞类型。目标 4. 确定 JCR 大鼠中组成型活性 p38 MAP 激酶的表达将恢复附带生长的细胞类型。拟议的研究采用多方面的方法来解决体内细胞特异性信号传导问题，通过采用技术来确定蛋白质表达、硫醇氧化和ROS产生的细胞特异性变化，并最终将这些测量结果与冠状动脉侧枝生长联系起来。这些研究将深入了解 ROS 和氧化还原信号调节冠状动脉侧枝生长的细胞特异性位置。公共卫生相关性：缺血性心脏病 (IHD) 仍然是美国的主要原因之一，也是美国医疗保健系统的主要成本。心脏中是否存在发育良好的侧支循环对 IHD 的发病率和死亡率具有巨大影响。与侧枝循环发育不良（低电导或侧枝生长证据）的患者相比，侧枝循环发育良好的患者猝死发生率较低，发生闭塞时梗塞面积较小，且预后较好。还值得注意的是，约 40% 的 IHD 患者表现出很少甚至没有冠状动脉侧支循环。人们还认识到，IHD的许多危险因素，例如高血压、肥胖、血脂异常，也会产生氧化应激，对冠状动脉侧枝生长产生负面影响。然而，发生腐败的细胞类型，例如氧化应激是否会破坏心肌细胞生长因子的产生，或者是否会破坏内皮细胞对生长因子的反应性，仍然未知。该提案的目标是描绘心脏中氧化还原信号和氧化应激对冠状动脉侧枝生长至关重要的细胞类型。该项目的最终目标是为更定向的治疗提供基础，以刺激 IHD 患者冠状动脉的生长。