Endoplasmic Reticulum Thiol Redox State and Unfolded Protein Response in Aging

描述（由申请人提供）：我的长期目标是专注于了解衰老的基本机制，我坚定地致力于作为一名独立研究者追求学术生涯。这份提案描述了一个全面的5年培训计划，为我的职业发展和向老龄化研究领域的终身教职过渡。该提案的指导阶段将在Vadim Gladyshev博士（氧化还原生物学和比较基因组学方面的知名专家）和Gary Ruvkun博士（衰老、遗传学和代谢生物学领域的先驱研究员）的指导下进行。此外，由具有内质网未折叠蛋白应激反应、氧化还原信号和荧光激活细胞分选分析专业知识的知名医学科学家组成的咨询小组。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里格姆妇女医院和哈佛医学院进行，这些医院提供了良好的研究和培训环境。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年龄相关疾病（如糖尿病、癌症和神经变性）的发病机制中发挥重要作用。细胞通过激活一种被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蛋白、抑制翻译、促进蛋白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与年龄相关疾病的病理生理有关的一个因素，但UPR信号在调节寿命中的作用尚不清楚。这一提议将验证调节UPR信号，无论是药理学还是遗传学，都可以激活保护性细胞应激反应并介导寿命延长的假设。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”内质网应激导致UPR信号的组成性上调，从而延长了出芽酵母的寿命。我们还发现，这些内质网/分泌途径突变体的寿命延长依赖于功能性内质网应激传感器蛋白Ire1p，并与内质网高氧化有关，这表明内质网的氧化还原状态与UPR信号密切相关，是酿酒葡萄球菌寿命的重要决定因素。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）内质网中蛋白质折叠能力和氧化还原状态如何随年龄变化？（iii）是否可以用小分子化合物调节UPR来提高内质网抗逆性和调节老化过程？

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?