p66Shc Signaling Functions in Cardiomyocytes

p66Shc 心肌细胞中的信号传导功能

基本信息

批准号：
8452689
负责人：
Susan F Steinberg
金额：
$ 37.93万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-15 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8452689
关键词：
Ablation Adrenergic Receptor Animal Model Antioxidants Apoptosis Apoptotic Attention Awareness Binding Biochemical Cardiac Cardiac Myocytes Cardiomyopathies Cardiovascular Diseases Cardiovascular system Caveolae Cell Death Cells Data Deterioration Development Etiology Event Evolution Failure Functional disorder Gene Expression Gene Silencing Generations Genes Genetic Goals Growth Heart Heart Diseases Heart Hypertrophy Heart failure Hypertrophy Knock-out Lead Link Longevity Measurable Mitochondria Modeling Modification Molecular Mus Natural History Oxidation-Reduction Oxidative Stress Pathogenesis Pathway interactions Phenotype Phosphorylation Predisposition Production Property Protein Isoforms Protein Kinase C Proteins Proto-Oncogene Proteins c-akt RNA Interference Reactive Oxygen Species Regulation Regulatory Pathway Relative (related person)Role Signal Transduction Small Interfering RNA Stimulus Testing Therapeutic adapter protein adenoviral-mediated aging gene base biological adaptation to stress diabetic in vivo novel novel therapeutics overexpression p66(ShcA) protein prevent protein structure public health relevance receptor response

项目摘要

DESCRIPTION (provided by applicant): p66Shc has recently emerged as a master regulator of reactive oxygen species (ROS) production and cardiovascular oxidative stress responses. While a role for p66Shc as an 'aging' gene that amplifies ROS generation in mitochondria and leads to measurable changes in lifespan in animal models is convincing, there is almost no information on p66Shc functions in cardiomyocytes. Preliminary studies in this application implicate p66Shc as a target of an adrenergic receptor pathway involving ROS and PKC that is localized to caveolae and regulates AKT-FOXO3a phosphorylation and anti-oxidant gene expression. We identify an effect of the G1q-dependent hypertrophy pathway to increase p66Shc expression and target p66Shc to mitochondria where it is predicted to increase ROS generation. We identify a role for protein kinase C-4 (PKC4) and PKC5 to influence p66Shc phosphorylation and localization. p66Shc regulation by PKC4 is particularly noteworthy, as PKC4 also participates in mitochondrial events that control ROS accumulation and apoptosis. Studies in this application will consider the role of p66Shc as a G1q-induced gene product that cooperates with PKC to influence the evolution of cardiac failure phenotypes. Specific aim I will identify p66Shc activation mechanisms and p66Shc signaling functions in cardiomyocytes. The goal is to determine whether p66Shc participates in ROS-regulatory signaling mechanisms in caveolae and mitochondria that influence growth and/or apoptosis. Most studies do not resolve the signaling functions of p66Shc versus the smaller p46/p52Shc isoforms (that do not regulate Redox signaling). Studies in Specific Aim II will use p66Shc knockout models, siRNA gene silencing, and adenoviral mediated overexpression strategies to resolve the specific functions of p66Shc (and critical determinants within the p66Shc protein structure) in mechanisms that regulate ROS production and cardiomyocyte growth/apoptosis. Studies in Specific Aim III will focus on the role of PKC4 to cooperate with p66Shc to control cardiomyocyte remodeling. These studies are based upon preliminary data showing that PKC4 is a p66Shc binding partner that influences p66Shc-S36 phosphorylation and p66Shc subcellular targeting. Finally, studies in Specific Aim IV will examine whether p66Shc gene silencing prevents the structural remodeling and functional deterioration that develops in selected models of cardiac hypertrophy/failure. Evidence that p66Shc contributes to the etiology and/or pathogenesis of cardiac dysfunction would provide the basis for future research focusing on p66Shc as an 'aging gene' that influences the evolution of cardiovascular disorders associated with oxidative stress (ischemic, diabetic, atherosclerotic, hypertensive, and hypertrophic heart disease). The long-term goal of such studies would be to develop novel therapeutic strategies that prevent p66Shc expression or interdict p66Shc function that afford cardioprotection.

描述（由申请人提供）：p66Shc 最近已成为活性氧（ROS）产生和心血管氧化应激反应的主要调节剂。虽然 p66Shc 作为一种“衰老”基因，可以放大线粒体中 ROS 的产生，并导致动物模型中寿命发生可测量的变化，这一作用令人信服，但几乎没有关于 p66Shc 在心肌细胞中功能的信息。本申请中的初步研究表明，p66Shc 是涉及 ROS 和 PKC 的肾上腺素受体途径的靶标，该途径定位于小凹并调节 AKT-FOXO3a 磷酸化和抗氧化基因表达。我们确定了 G1q 依赖性肥大途径对增加 p66Shc 表达的影响，并将 p66Shc 靶向线粒体，预计在线粒体中会增加 ROS 的产生。我们确定了蛋白激酶 C-4 (PKC4) 和 PKC5 在影响 p66Shc 磷酸化和定位方面的作用。 PKC4 对 p66Shc 的调节尤其值得注意，因为 PKC4 还参与控制 ROS 积累和细胞凋亡的线粒体事件。本申请中的研究将考虑 p66Shc 作为 G1q 诱导基因产物的作用，它与 PKC 合作影响心力衰竭表型的进化。具体目标我将确定心肌细胞中的 p66Shc 激活机制和 p66Shc 信号传导功能。目标是确定 p66Shc 是否参与小窝和线粒体中影响生长和/或凋亡的 ROS 调节信号机制。大多数研究并未解析 p66Shc 与较小的 p46/p52Shc 亚型（不调节氧化还原信号传导）的信号传导功能。 Specific Aim II 的研究将使用 p66Shc 敲除模型、siRNA 基因沉默和腺病毒介导的过表达策略来解决 p66Shc（以及 p66Shc 蛋白质结构内的关键决定因素）在调节 ROS 产生和心肌细胞生长/凋亡的机制中的特定功能。 Specific Aim III的研究将重点关注PKC4与p66Shc配合控制心肌细胞重塑的作用。这些研究基于初步数据，表明 PKC4 是 p66Shc 结合伴侣，影响 p66Shc-S36 磷酸化和 p66Shc 亚细胞靶向。最后，Specific Aim IV 中的研究将检验 p66Shc 基因沉默是否可以防止选定的心脏肥大/衰竭模型中发生的结构重塑和功能恶化。 p66Shc 与心脏功能障碍的病因学和/或发病机制有关的证据将为未来的研究奠定基础，重点关注 p66Shc 作为一种“衰老基因”，影响与氧化应激相关的心血管疾病（缺血性、糖尿病、动脉粥样硬化、高血压、和肥厚性心脏病）。此类研究的长期目标是开发新的治疗策略，阻止 p66Shc 表达或阻断 p66Shc 功能，从而提供心脏保护作用。