Reductive Stress Induces Proteotoxic Cardiac Disease

还原性压力诱发蛋白毒性心脏病

• 批准号: 9751933
• 负责人: Rajasekaran Namakkal Soorappan
• 金额: $ 58.02万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-29 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751933
• 关键词: Adaptor Signaling Protein; Address; Adverse effects; Alzheimer' s Disease; Animal Model; Antioxidants; Automobile Driving; Autophagocytosis; Autophagosome; Biochemical; Biochemistry; Biological Assay; Biology; Cardiac; Cardiovascular system; Cell Nucleus; Cessation of life; Chronic; Clinical; Clinical Trials; Collaborations; Comorbidity; Computer Analysis; Coupled; Cytosol; DNA Sequence Alteration; Disease Progression; Etiology; Event; Exhibits; Functional disorder; Genes; Genetic; Genetic Predisposition to Disease; Glutathione Disulfide; Goals; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; High Pressure Liquid Chromatography; Human; Hypertrophic Cardiomyopathy; Impairment; Injury; Label; Laboratories; Lead; Link; Lipid Peroxidation; Measures; Mediating; Molecular; Molecular Biology; Molecular Genetics; Molecular Profiling; Mus; Mutation; Myocardial; Myocardial dysfunction; Myocardium; Organ; Oxidation-Reduction; Oxidative Stress; Participant; Pathogenesis; Pathologic; Pathway interactions; Patients; Phenotype; Physicians; Post-Translational Protein Processing; Process; Property; Protein Dynamics; Proteins; Proteome; Proteomics; Quality Control; Reduced Glutathione; Regulation; Regulatory Pathway; Research; Role; Sample Size; Scientist; Signal Transduction; Stress; Supplementation; Technology; Testing; Therapeutic; Time; Ubiquitination; base; computational platform; endoplasmic reticulum stress; experience; genetic variant; misfolded protein; mouse model; multidisciplinary; novel; patient subsets; peripheral blood; personalized therapeutic; premature; prevent; programs; protein aggregation; protein degradation; protein folding; proteotoxicity; response; sensor; stress reduction; targeted sequencing

Conventionally, oxidative stress is considered pathological to cardiac protein quality control (PQC), which rationalizes the therapeutic potential of antioxidants. However, many clinical trials of antioxidant supplementation failed to deliver a positive impact and, rather, demonstrated adverse effects in various organs, including those in the cardiovascular system. Emerging evidence suggests that reductive stress (RS), also known as antioxidative stress, may cause ER stress and accumulation of mis/unfolded proteins. To fully comprehend the molecular interplay underlying RS-mediated proteotoxicity and the time frame for myocardium experiencing a transition from adaptive to maladaptive remodeling, it is critical to identify the molecular participants, their dynamic interplay, and resulting sequential events (e.g., autophagy) that regulate PQC. Recently, our exciting and novel clinical observations revealed a link of chronic RS (cRS) underlying disease progression of human heart failure (HF). We screened a selected group of HF patients (n=50, without other major comorbidities) for their peripheral blood redox state; among them, a subset (n=8) displayed the RS condition. Our proposed study will entail a translational component utilizing proteomics approaches and molecular biology to better understand the etiology of RS in mouse models and to identify its relevance in HF patients. Our central hypothesis is that cRS will alter proteome properties (e.g., protein dynamics & post- translational modifications, or PTMs) and damage autophagy signaling, leading to persistent proteotoxicity and cardiac dysfunction, thereby driving maladaptive remodeling in animal models and in human heart diseases. We propose three aims: Aim 1 will determine altered protein dynamics, redistributed PTMs, and perturbed autophagy subproteome in RS conditions. We will define the “reductome signatures” in control and cRS phenotypes. Aim 2 will examine the impact of cRS on progressive damage of autophagy that may lead to insufficient cargo-clearance and proteotoxicity in the myocardium over time. We will assess autophagosome formation, autophagy flux, and protein folding capacity under cRS conditions and examine whether enhancing autophagy delays and/or prevents proteotoxicity in mice. Aim 3 will examine the “redox phenotype” in the peripheral blood of HF patients using HPLC based quantification of (a) GSH redox ratio, (b) lipid peroxidation, and (c) total antioxidant capacity, as well as extract the molecular “reductome signatures” in HF patients with RS using a computational platform to determine essential proteome features and regulatory pathways. This aim will establish a translational value for the RS hypothesis in human HF. We have assembled a multidisciplinary team (scientists & physicians) with documented expertise in redox biology, biochemistry, proteomics, and computational analyses. The genetic mouse models of RS, the technology platforms, and the biochemical assays to evaluate RS in mouse and in human are all established in our laboratories. We anticipate the successful completion of our proposed goals.

通常，氧化应激被认为是对心脏蛋白质质量控​​制（PQC）的病理学，这 合理化抗氧化剂的治疗潜力。但是，许多抗氧化剂的临床试验 补充未能产生积极的影响，而是在各种器官中表现出不利影响， 包括心血管系统中的那些。新兴证据表明，压力减轻（RS）也 称为抗氧化应激，可能会导致MIS/展开蛋白质的ER应力和积累。完全 理解分子相互作用的基础RS介导的蛋白毒性和心肌的时间范围 经历从适应性重塑到不良适应性重塑的过渡，识别分子至关重要 参与者，他们的动态相互作用以及调节PQC的结果顺序事件（例如自噬）。 最近，我们令人兴奋和新颖的临床观察结果揭示了慢性RS（CRS）的联系 人类心力衰竭的疾病进展（HF）。我们筛选了一组选定的HF患者（n = 50，没有 其他主要合并症）用于外周血氧化还原状态；其中，一个子集（n = 8）显示了RS 健康）状况。我们拟议的研究将需要使用蛋白质组学方法进行翻译的组件，并 分子生物学，以更好地了解小鼠模型中RS的病因并确定其在HF中的相关性 患者。我们的中心假设是CRS将改变蛋白质特性（例如，蛋白质动力学和后 翻译修改或PTMS）和损害自噬信号传导，导致持续的蛋白质毒性和 心脏功能障碍，从而在动物模型和人类心脏病中驱动适应不良的重塑。 我们提出三个目标：AIM 1将决定改变的蛋白质动力学，重新分布的PTM和扰动 RS条件下的自噬副蛋白酶。我们将在控制和CRS中定义“还原签名” 表型。 AIM 2将检查CRS对自噬进行性损害的影响，这可能导致 随着时间的推移，心肌的货物清除率和蛋白质毒性不足。我们将评估自噬体 在CRS条件下形成，自噬通量和蛋白质折叠能力，并检查是否增强 自噬延迟和/或阻止小鼠的蛋白质毒性。 AIM 3将检查中的“氧化还原表型” 使用基于HPLC的（a）GSH氧化还原比，（b）脂质过氧化的基于HFC患者的外周血 （c）总抗氧化能力，以及提取HF患者的分子“还原标志” RS使用计算平台来确定必需的蛋白质组特征和调节途径。这 AIM将在人HF中为RS假设建立翻译价值。 我们已经组建了一个多学科团队（科学家和医生），并在氧化还原方面拥有有记录的专业知识 生物学，生物化学，蛋白质组学和计算分析。 Rs的遗传小鼠模型， 技术平台以及用于评估鼠标和人类RS的生化测定法 我们的实验室。我们预计我们提出的目标将成功完成。