Endoplasmic Reticulum Thiol Redox State and Unfolded Protein Response in Aging

衰老过程中内质网硫醇氧化还原状态和未折叠蛋白反应

• 批准号: 8549048
• 负责人: Vyacheslav M Labunskyy
• 金额: $ 13.15万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8549048
• 关键词: Address; Age; Aging; Aging-Related Process; Animal Model; Biology; Biology of Aging; Caenorhabditis elegans; Cell Aging; Cells; Cellular Stress; Cellular Stress Response; Chemicals; Collection; Commit; DNA Microarray Chip; Data; Development; Diabetes Mellitus; Disease; Disulfides; Dithiothreitol; Endoplasmic Reticulum; Environment; Eukaryotic Cell; Faculty; Fluorescence-Activated Cell Sorting; Functional disorder; Gene Deletion; Genes; Genetic; Genetic Engineering; Genetic Screening; Goals; Heat shock proteins; Homeostasis; Hospitals; Human; Hydrogen Peroxide; Hypoxia; Individual; Lead; Link; Longevity; Malignant Neoplasms; Measures; Mediating; Medical; Mentors; Mentorship; Metabolism; Microarray Analysis; Modification; Molecular Chaperones; Mutation; Nerve Degeneration; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phase; Play; Positioning Attribute; Protein Secretion; Proteins; Quality Control; RNA Splicing; Reporter; Research; Research Personnel; Research Training; Resistance; Role; Saccharomyces cerevisiae; Saccharomycetales; Scientist; Signal Pathway; Signal Transduction; Stress; Stress Response Signaling; Sulfhydryl Compounds; Testing; Training Programs; Translations; Tunicamycin; Up-Regulation; Woman; Yeasts; age related; aged; base; biological adaptation to stress; career; career development; cell age; cell type; chemical genetics; comparative genomics; disulfide bond; endoplasmic reticulum stress; high throughput screening; improved; in vivo; inhibitor/antagonist; insight; longevity gene; medical schools; mutant; novel; novel therapeutics; nucleotide analog; overexpression; prevent; protein aggregation; protein folding; protein misfolding; response; sensor; small molecule; stress protein; transcription factor

DESCRIPTION (provided by applicant): My long-term goals are focused on understanding the basic mechanisms of aging and I am strongly committed to pursuing an academic career as an independent investigator. This proposal describes a comprehensive 5-year training program for my career development and transition to a tenure-track faculty position in the field of aging research. The mentored phase of the proposal will be carried out under the mentorship of Dr. Vadim Gladyshev, a well-established expert in redox biology and comparative genomics, and the co-mentorship of Dr. Gary Ruvkun, a pioneering researcher in the biology of aging, genetics and metabolism. Additionally, an advisory panel of established medical scientists with expertise in the ER unfolded protein stress response, redox signaling, and fluorescence activated cell sorting analysis consisting of Drs. Joseph Loscalzo, Gokhan Hotamisligil and Ronglih Liao will provide further scientific and career guidance. The planned career development activities will be carried out at the Brigham and Women's Hospital and Harvard Medical School, which provide an excellent research and training environment. Research plan: ER stress and protein misfolding have been shown to play an important role in aging and pathogenesis of various age-related diseases, such as diabetes, cancer and neurodegeneration. Cells adapt to accumulation of misfolded proteins in the ER by activating an evolutionary conserved protective mechanism known as the unfolded protein response (UPR). This signaling pathway restores ER homeostasis by degrading misfolded proteins, inhibiting translation, and facilitating protein folding and secretion. Although UPR dysfunction is increasingly recognized as a contributing factor to the pathophysiology of age-related diseases, the role of UPR signaling in regulating lifespan is not known. This proposal will test the hypothesis that modulating UPR signaling, either pharmacologically or genetically, can activate protective cellular stress responses and mediate lifespan extension. Our preliminary data demonstrate that constitutive up-regulation of the UPR signaling due to "mild" ER stress caused by selective inactivation of individual protein folding and maturation factors in the ER leads to increased longevity in budding yeast. We also found that extended lifespan in these ER/secretory pathway mutants is dependent on functional ER stress sensor protein, Ire1p, and is associated with ER hyperoxidation, suggesting that redox status in the ER is closely interlinked with the UPR signaling and is an important determinant of S. cerevisiae lifespan. Building upon these findings, we propose to address the following specific questions: (i) What are the mechanisms by which UPR and ER redox state regulate longevity? (ii) How protein folding capacity and redox state in the ER change with age? (iii) Can modulation of the UPR with small molecule compounds be used to improve ER stress resistance and regulate aging process?

描述（由申请人提供）：我的长期目标集中在了解衰老的基本机制，并且我坚定地致力于作为一名独立研究者追求学术生涯。该提案描述了一个为期 5 年的综合培训计划，旨在促进我的职业发展和向衰老研究领域的终身教授职位的过渡。该提案的指导阶段将在氧化还原生物学和比较基因组学领域的知名专家 Vadim Gladyshev 博士和衰老生物学先驱研究员 Gary Ruvkun 博士的共同指导下进行、遗传学和新陈代谢。此外，由在 ER 未折叠蛋白应激反应、氧化还原信号传导和荧光激活细胞分选分析方面具有专业知识的资深医学科学家组成的顾问小组由 Drs. Joseph Loscalzo、Gokhan Hotamisligil 和 Ronglih Liao 将提供进一步的科学和职业指导。计划中的职业发展活动将在布莱根妇女医院和哈佛医学院进行，那里提供了优良的研究和培训环境。研究计划：内质网应激和蛋白质错误折叠已被证明在衰老和各种与年龄相关的疾病（如糖尿病、癌症和神经退行性疾病）的发病机制中发挥着重要作用。细胞通过激活称为未折叠蛋白反应（UPR）的进化保守保护机制来适应内质网中错误折叠蛋白的积累。该信号通路通过降解错误折叠的蛋白质、抑制翻译以及促进蛋白质折叠和分泌来恢复内质网稳态。尽管UPR功能障碍越来越被认为是年龄相关疾病病理生理学的一个促成因素，但UPR信号在调节寿命中的作用尚不清楚。该提案将检验以下假设：通过药理学或遗传手段调节 UPR 信号传导可以激活保护性细胞应激反应并介导寿命延长。我们的初步数据表明，由于内质网中单个蛋白质折叠和成熟因子的选择性失活引起的“轻度”内质网应激，UPR信号的组成性上调会导致芽殖酵母的寿命延长。我们还发现，这些 ER/分泌途径突变体的寿命延长依赖于功能性 ER 应激传感器蛋白 Ire1p，并且与 ER 过度氧化相关，这表明 ER 中的氧化还原状态与 UPR 信号密切相关，是一个重要的决定因素。酿酒酵母的寿命。基于这些发现，我们建议解决以下具体问题：（i）UPR和ER氧化还原状态调节寿命的机制是什么？ (ii) ER 中的蛋白质折叠能力和氧化还原状态如何随年龄变化？ (iii) 用小分子化合物调节 UPR 是否可以用来提高 ER 应激抵抗力并调节衰老过程？