Mechanisms of Cardioprotection

心脏保护机制

基本信息

批准号：
7248566
负责人：
Roberto Bolli
金额：
$ 55.8万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-07-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7248566
关键词：
Ablation Analysis of Variance Animals Apoptosis Arts Biochemical Genetics Biochemistry Biological Assay Cardiac Cardiovascular system Catabolism Class Confocal Microscopy Coronary Arteriosclerosis Coronary Occlusions Cyclic GMP Cytoprotection Data Development Diethyldithiocarbamate Disruption Dose EMSA Enzyme Immunoassay Exercise Gene Targeting Gene Transfer Techniques Generations Genes Genetic Genetic Techniques Genetically Engineered Mouse HIF1A gene Heart Heme Hypoxia Inducible Factor I Kappa B-Alpha I-kappa B Proteins Immunohistochemistry Individual Infarction Inhibition of Apoptosis Interferon Activation Investigation Ischemia Janus kinase Knock-out Knockout Mice Late Effects Lead Left ventricular structure Lipopolysaccharides Measures Mediating Mediator of activation protein Modeling Molecular Molecular Biology Molecular Genetics Multiprotein Complexes Mus Mutant Strains Mice Myocardial Ischemia Myocardium Myosin Heavy Chains N-acetylpenicillamine NF-kappa B Necrosis Neuro-Oncological Ventral Antigen 2 Nitric Oxide Synthase Nitroglycerin Nuclear PTGS2 gene Pathologic Pathology Patients Pharmaceutical Preparations Phase Phenotype Physiological Physiological reperfusion Physiology Play Polymerase Chain Reaction Prevention Process Property Prostaglandin-Endoperoxide Synthase Prostaglandins Protein Chemistry Protein Kinase C Protein Overexpression Proteins Recruitment Activity Regulation Relative (related person)Reperfusion Injury Reperfusion Therapy Repression Resistance Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription Ribonucleases Role S-nitro-N-acetylpenicillamine STAT1 gene STAT3 gene Signal Transduction Site Stimulus Stress Structure Supplementation TNFRSF5 gene Techniques Testing Therapeutic Thromboxane B2 Time Time Study Tissues Transcription Coactivator Transgenic Mice Transgenic Organisms UC01 Up-Regulation Ventricular Cardiac alpha-Myosin Waste Products cell type concept cyclooxygenase 1 cyclooxygenase 2 diethylenetriamine dimer heat shock transcription factor heme oxygenase-1 in vivo inhibitor/antagonist insight interdisciplinary approach knockout gene mutant novel preconditioning progesterone 11-hemisuccinate-(2-iodohistamine)protective effect response size therapeutic gene transcription factor tumor

项目摘要

DESCRIPTION (provided by applicant): We have recently found that HO-1 serves as an obligatory mediator of three different forms of late preconditioning (PC), suggesting a central cardioprotective role of this protein. Despite this, and despite the fact that HO-1 is one of the most frequently induced proteins known to exist, the regulation of cardiac HO-1 and the mechanism of its cardioprotective actions are unknown. The objective of this proposal is to elucidate the molecular mechanisms underlying the newly discovered cardioprotective role of HO-1 and to use this knowledqe to develop novel therapies. We will test the hypothesis that diverse PC stimuli upregulate HO-1 via eNOS-dependent generation of NO and subsequent activation of a cluster of stress-responsive transcription factors, and that the salubrious effects of HO-1 are due to CO-dependent inhibition of apoptosis and modulation of other co-induced proteins (iNOS, COX-2). We propose that CO, far from being a toxic waste product, plays a fundamental role in protecting the heart against ischemia and apoptosis. Three different forms of late PC elicited by a pathological (ischemia), physiological (exercise), and pharmacological (NO donor) stimulus will be interrogated in an effort to develop a comprehensive pathophysiologic paradigm. A broad multidisciplinary approach will be used that will combine diverse techniques (integrative physiology, protein chemistry, biochemistry, pathology, confocal microscopy, immunohistochemistry, molecular biology, gene targeting, and transgenesis) and will integrate genetic and biochemical information at the molecular level with physiological information at the whole animal level. A total of 16 genetically engineered mouse lines will be studied. The effects of late PC and HO-1 on infarct size, apoptosis, and necrosis will be determined using HO-1-/- and HO-1 transgenic mice in conjunction with state-of-the-art pathology, immunohistochemistry, and confocal microscopy techniques. The effects of CO on myocardial ischemia/reperfusion injury and apoptosis will be determined by using a novel class of compounds (CO-releasing molecules) that have potential therapeutic applications. The role of specific transcription factors (NF-kappaB, HIF-1alpha, HSF1, STAT1, and STAT3) in HO-1 upregulation will be established by the use of a novel transdominant IkappaBalpha mutant mouse with cardiac-specific repression of NF-kappaB activity, knockouts for HIF-1alpha, HSF1, and three specific NF-kappaB subunits (p50, RelB, and c-Rel), and a novel conditional STAT3 knockout mouse. The functional interactions of HO-1 with the other two co-mediators of late PC (iNOS and COX-2) will be deciphered in vivo by cardiac-specific overexpression and targeted disruption of each of these three genes. This project should produce important new insights into the role of apoptosis vis-a-vis necrosis in late PC and the mechanism of HO-1-dependent cytoprotection. If the studies of CO donors confirm our preliminary data, they may pave the way for the development of novel antiischemic and antiapoptotic therapies predicated upon the supplementation of exogenous CO to patients with coronary artery disease.

描述（由申请人提供）：我们最近发现，HO-1是三种不同形式的晚期预处理（PC）的强制性介体，这表明该蛋白质的中央心脏保护作用。尽管如此，尽管HO-1是已知存在的蛋白质之一，但心脏HO-1的调节及其心脏保护作用的机制尚不清楚。该提案的目的是阐明HO-1新发现的心脏保护作用的分子机制，并使用此知识来开发新的疗法。我们将测试以下假设：不同的PC刺激通过eNOS依赖性的NO和随后激活一组应力反应转录因子上调HO-1，而HO-1的作用是由于共依赖性抑制了凋亡和其他共聚蛋白（Inos，Cox，Cox，Cox，Cox-cox-cox）的共依赖性抑制。我们建议，远非有毒废物的CO在保护心脏免受缺血和凋亡中起着基本作用。病理学（缺血），生理（运动）和药理学（无供体）刺激的三种不同形式的PC的三种形式将受到询问，以开发全面的病理生理范式。将使用一种广泛的多学科方法，该方法将结合多种技术（综合生理学，蛋白质化学，生物化学，病理学，共聚焦显微镜，免疫组织化学，分子生物学，基因靶向和转基因），并将与分子级别整合在物理级别的分子级别。总共研究了16种基因工程的小鼠系。 PC和HO-1晚期对梗塞大小，凋亡和坏死的影响将使用HO-1 - / - 和HO-1转基因小鼠与最先进的病理学，免疫组织化学以及共斑显微镜技术结合使用。 CO对具有潜在治疗应用的新型化合物（共释放分子）的新型化合物（共释放分子）来确定CO对心肌缺血/再灌注损伤和凋亡的影响。特定转录因子（NF-KAPPAB，HIF-1ALPHA，HSF1，STAT1和Stat3）在HO-1上调中的作用将通过使用新型的跨型Ikappabalpha突变小鼠使用心脏抑制NF-kappab活性，nf-kappab活性，HIF-1Alpha，HSF1和三个特定的NF--（PAP-1-app-abp）（PAP-1），从而确定。 C-Rel）和一种新颖的有条件STAT3敲除鼠标。 HO-1与晚期PC（INOS和COX-2）的其他两个联合中等体的功能相互作用将通过心脏特异性的过表达在体内解密，并靶向这三个基因中的每一个。该项目应该对PC晚期凋亡性凋亡的作用以及HO-1依赖性细胞保护的机理产生重要的新见解。如果对CO捐赠者的研究证实了我们的初步数据，则它们可能为发展为冠状动脉疾病患者补充外源性CO的新型抗疾病和抗凋亡疗法的发展铺平道路。