权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Role of p67phox in myocardial hypertrophy

p67phox 在心肌肥厚中的作用

基本信息

批准号：
7683210
负责人：
Lei Xiao
金额：
$ 35.62万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-03 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7683210
关键词：
Adrenergic Agents Adrenergic Receptor Adult Age Animal Model Blood Vessels Breeding Cardiac Cardiac Myocytes Cardiovascular Diseases Chronic Congestive Heart Failure Crossbreeding Data Development Dominant-Negative Mutation Enzymes Extracellular Signal Regulated Kinases Gender Generations Genes Goals Heart Heart Hypertrophy Heart failure Human Hybrids Hypertrophy In Vitro Laboratories Lead MEKs Measures Mechanical Stress Mechanics Mediating Mitogen-Activated Protein Kinases Modeling Molecular Mus Muscle Cells Myocardial Myocardium NADPH Oxidase Neonatal Operative Surgical Procedures Oxidases Parents Phagocytes Phenotype Phosphotransferases Play Production Protein Biosynthesis Proteins Public Health Rattus Reactive Oxygen Species Reporting Research Research Personnel Role Scaffolding Protein Signal Transduction Smooth Muscle Myocytes Stimulus Stretching System Testing Transgenes Transgenic Mice Transgenic Organisms Ventricular Wild Type Mouse adrenergic aorta constriction ascending aorta base disability enzyme activity fetal hemodynamics in vivo inhibitor/antagonist mortality mouse model mutant neutrophil cytosol factor 67K novel offspring overexpression pressure prevent protein protein interaction public health relevance

项目摘要

DESCRIPTION (provided by applicant): NADPH oxidase (NOX) appears to play a role in the development of myocardial hypertrophy and subsequent progression of heart failure. However, the signaling mechanisms of NOX in cardiac myocytes is unknown. The long-term goal of this research is to understand the signaling mechnisms of NOX in the development of myocardial hypertrophy and heart failure induced by excessive adrenergic stimulation and hemodynamic pressure overload. Our prelminary data indicate that p67phox, the major cytosolic subunit of NOX, mediates the development of myocardial hypertrophy through a novel reactive oxygen species (ROS) - independent mechanism. Based on these data, we hypothesize that NOX/p67phox mediates myocardial hypertrophy that is induced by hypertrophic stimuli including 11-AR-stimulation and hemodynamic pressure overload, and its signaling function involve a novel ROS-independent mechanism. To test this hypothesis, we proposed the following three Specific Aims: In Specific Aim 1, we will determine whether NOX/p67phox mediates 11-adrenergic receptor (11-AR)- stimulated myocardial hypertrophy and whether this involves a novel ROS-independent signaling mechanism in vitro. Dominant negative (DN) mutants and wild type (WT) contstructs of two NOX cytosolic subunits, p67phox and p47phox. Mutants or WT constructs will be overexpressed in cultured adult rat ventricular myocytes (ARVM). The effects of these mutants or WT constructs on 11-AR stimulation-induced hypertrophy and NOX enzyme activity will be determined. The potential protein-protein interaction between p67phox and MEK-ERK cascade will also be determined in this aim. In Specific Aim 2, we will test if p67phox mediates 11-AR-stimulated cardiac hypertrophy in vivo. We will use the newly generated heterozygous DN-p67 transgenic (TG) mice with cardiac-specific overexpression of the DN p67phox mutant to generate homozygous DN-p67 TG mice. Since the commercially available TG mice with cardiac-specific overexpression of a constitutively activated mutant of 11B-AR (CAM-11B-AR) develop cardiac hypertrophy, we will crossbreed these two TG lines (DN-p67 and CAM-11B-AR) to study the effect of overexperssion of DN p67phox mutant protein in the heart on 11B-AR-induced myocardial hypertrophy in vivo. In Specific Aim 3, we will test the role of myocardial p67phox in mediating hemodynamic overload- induced myocardial hypertrophy in vivo. The homozygous DN-p67 TG mice and age- and gender-matched WT control mice will be subjected to chronic pressure overload caused by ascending aorta constriction (AAC). The cardiac phenotypes and function of these TG and WT mice with AAC or sham operation will be studied to determine the role of p67phox in mediating chronic pressure overload-induced myocardial hypertrophy in vivo. Our studies will provide new understanding of the role and novel mechanisms of NOX/p67phox in the development of cardiac hypertrophy that should lead to novel treatments for congestive heart failure. PUBLIC HEALTH RELEVANCE Our studies will provide new understanding of the novel pathological mechanisms of p67phox and NADPH oxidase in the development of hypertrophy in the heart, which could lead to the major cardiovascular disease, heart failure. Therefore, our research will have direct relevance to a common cause of heart failure, which has a high mortality and disability rate and is a major threat to the public health.

描述（由申请方提供）：NADPH氧化酶（NOX）似乎在心肌肥大的发生和随后的心力衰竭进展中发挥作用。然而，NOX在心肌细胞中的信号转导机制尚不清楚。本研究的长期目标是了解NOX在过度肾上腺素能刺激和血流动力学压力超负荷引起的心肌肥厚和心力衰竭的发展中的信号机制。我们的初步数据表明，p67 phox，主要的胞浆亚基的NOX，介导的发展心肌肥厚通过一种新的活性氧（ROS）的非依赖性机制。基于这些数据，我们假设，NOX/p67 phox介导的心肌肥厚，是由肥大刺激，包括11-AR刺激和血流动力学压力过载，其信号转导功能涉及一种新的ROS非依赖性机制。为了验证这一假设，我们提出了以下三个具体目标：在具体目标1中，我们将确定NOX/p67 phox是否介导11-肾上腺素能受体（11-AR）刺激的心肌肥大，以及这是否涉及一种新的ROS非依赖性信号转导机制。显性阴性（DN）突变体和野生型（WT）构成两个NOX胞质亚基，p67 phox和p47 phox。突变体或WT构建体将在培养的成年大鼠心室肌细胞（ARVM）中过表达。将确定这些突变体或WT构建体对11-AR刺激诱导的肥大和NOX酶活性的影响。 p67 phox和MEK-ERK级联之间潜在的蛋白质-蛋白质相互作用也将在此目标中确定。在特定目标2中，我们将测试p67 phox是否介导体内11-AR刺激的心脏肥大。我们将使用新产生的具有DN p67 phox突变体的心脏特异性过表达的杂合DN-p67转基因（TG）小鼠来产生纯合DN-p67 TG小鼠。由于市售的TG小鼠与心脏特异性过表达的组成型激活的突变体11 B-AR（CAM-11 B-AR）发展心肌肥大，我们将这两个TG系（DN-p67和CAM-11 B-AR）杂交，以研究DN p67 phox突变蛋白在心脏中的过表达对11 B-AR诱导的心肌肥大的影响。在具体目标3中，我们将测试心肌p67 phox在体内介导血流动力学超负荷诱导的心肌肥大中的作用。纯合DN-p67 TG小鼠和年龄和性别匹配的WT对照小鼠将经受由升主动脉缩窄（AAC）引起的慢性压力超负荷。将研究这些患有AAC或假手术的TG和WT小鼠的心脏表型和功能，以确定p67 phox在体内介导慢性压力超负荷诱导的心肌肥大中的作用。我们的研究将提供新的理解的作用和新的机制，NOX/p67 phox在心肌肥厚的发展，应导致新的治疗充血性心力衰竭。公共卫生相关性我们的研究将为p67 phox和NADPH氧化酶在心脏肥大发展中的新病理机制提供新的理解，这可能导致主要的心血管疾病心力衰竭。因此，我们的研究将与心力衰竭的常见原因直接相关，心力衰竭具有高死亡率和致残率，是对公众健康的主要威胁。