DNA methylation of extracellular superoxide dismutase in pulmonary hypertension

肺动脉高压细胞外超氧化物歧化酶 DNA 甲基化

基本信息

批准号：
8335465
负责人：
Eva S. Nozik
金额：
$ 7.65万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-23 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8335465
关键词：
Address Adult Affect Animal Model Antioxidants Attenuated Azacitidine Biology Blood Vessels Bronchi Cell Proliferation Cells Child CpG Islands Cytosine DNA Methylation Data Development Diagnosis Disease Disease Progression Enzymes Epigenetic Process Event Fibrosis Foundations Freezing Gene Delivery Gene Expression Gene Proteins Genes Genetic Transcription Genomics Goals Guanosine Human Human Development Hypermethylation Hypertension Inflammation Injury Lead Life Lung Lung Transplantation Lung diseases Measures Mediating Messenger RNA Methylation Modification Molecular National Heart, Lung, and Blood Institute Normal Cell Nucleotides Organ Transplantation Pathogenesis Pathway interactions Patients Process Promoter Regions Protein Isoforms Proteins Pulmonary Circulation Pulmonary Hypertension Pulmonary artery structure Reactive Oxygen Species Regulation Research Research Personnel Research Proposals Role Sampling Serum Smooth Muscle Myocytes Superoxide Dismutase Superoxides Testing Therapeutic Time Tissue Banking Tissue Banks Tissues Transgenic Mice Vascular Diseases Vascular remodeling Work attenuation base bisulfite cancer cell demethylation disorder control enzyme activity extracellular human disease improved inhibitor/antagonist neoplastic cell novel novel therapeutics promoter protein expression pulmonary arterial hypertension tool

项目摘要

Idiopathic pulmonary arterial hypertension (IPAH) is a life-threatening disease of the pulmonary circulation affecting children and adults. Accumulating evidence indicates that one key factor that contributes to the pathogenesis of vascular diseases, including PAH, is an increase in reactive oxygen species, such as superoxide (O2-), that exceed antioxidant capabilities. One critically important antioxidant in the vessel wall is extracellular superoxide dismutase (EC-SOD), the sole enzymatic defense against extracellular O2-. EC-SOD is the most highly expressed SOD isoform in the vasculature under normal conditions, and EC-SOD expression is decreased in animal models of lung or vascular injury and several human diseases, including one study showing a decrease in EC-SOD protein in the bronchus of 8 patients with IPAH. Augmenting EC- SOD activity in transgenic mice or through adenoviral gene delivery attenuates pulmonary vascular remodeling and pulmonary hypertension. The regulation of EC-SOD in IPAH has not been investigated. It is now recognized that gene expression can be regulated by epigenetic modifications including cytosine methylation within the promoter region, specifically cytosines adjacent to guanosine nucleotides (CpG islands). New data identify CpG islands within the EC-SOD promoter that can be methylated, and indicate that hypermethylation of the EC-SOD promoter inhibits EC-SOD transcription in human cancer cells, contributing to enhanced tumor cell proliferation. Based on these data, we hypothesize that DNA methylation of the EC-SOD promoter mediates silencing of this protective gene and contributes to the pathogenesis of idiopathic pulmonary arterial hypertension. Aim 1 will utilize lung and pulmonary artery tissue and serum procured at the time of organ transplantation in patients with IPAH, disease-specific controls with PAH due to other causes, or patients without PAH to determine whether EC-SOD gene and protein as well as enzyme activity are decreased in IPAH and test the impact of EC-SOD expression on pulmonary artery smooth muscle cell proliferation. Specific Aim 2 will then use bisulfite genomic sequencing to test whether the EC-SOD promoter in pulmonary artery tissue and pulmonary artery smooth muscle cells from humans with IPAH is hypermethylated, and if reversal of methylation restores EC-SOD expression and normal cell proliferation. The tissue and cell samples used in this study will have been procured and processed through the efforts of the investigators of the Pulmonary Hypertension Breakthrough Initiative. Our findings will serve as strong preliminary data for the subsequent submission of a comprehensive RO1 application investigating the regulation of EC-SOD in IPAH. The long term goals are to establish new pathways important in the regulation of this pivotal antioxidant enzyme in the vessel wall, identify the role of extracellular superoxide in proliferation, inflammation and fibrosis contributing to pulmonary vascular remodeling, and provide a rationale for novel therapeutic tools to improve treatment of this lethal disease affecting children and adults.