Elucidating the role of ER remodeling in aging of C. elegans

阐明 ER 重塑在秀丽隐杆线虫衰老中的作用

• 批准号: 10536456
• 负责人: Eric KF Donahue
• 金额: $ 3.19万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10536456
• 关键词: Adult; Advisory Committees; Age; Age of Onset; Aging; Automobile Driving; Autophagocytosis; Biological Aging; Caenorhabditis elegans; Calcium; Caloric Restriction; Cell physiology; Cellular biology; Child; Chronic; Chronic Disease; Communication; Complement; Data; Developmental Cell Biology; Disease; Elderly; Electron Microscopy; Endoplasmic Reticulum; Environment; Environmental Risk Factor; Equilibrium; Eukaryota; Event; Family member; Feedback; Foundations; Genes; Genetic; Genetic Engineering; Homeostasis; Human; Impairment; Intervention; Laboratories; Laboratory Study; Licensing; Lipids; Longevity; Mediating; Mediator of activation protein; Membrane; Metabolism; Mitochondria; Molecular; Morbidity - disease rate; Morphology; Nature; Neurodegenerative Disorders; Nutrient; Organelles; Pathologic; Pathway interactions; Physiological; Physiology; Play; Population; Prevalence; Prevention; Process; Proteins; Proteome; Public Health; RNA Interference; Recording of previous events; Recycling; Regulation; Reporter; Ribosomes; Role; Rough endoplasmic reticulum; Shapes; Signal Transduction; Site; Smooth Endoplasmic Reticulum; Stress; Structure; Supervision; Techniques; Testing; Therapeutic; Time; Tubular formation; United States; Universities; Work; age related; base; detection of nutrient; dietary restriction; disability; experience; experimental study; healthspan; healthy aging; in vivo imaging; knock-down; loss of function; mortality; mutant; normal aging; novel; preservation; prevent; professor; protective effect; proteostasis; receptor; therapeutic evaluation; therapeutic target; tomography

PROJECT SUMMARY/ABSTRACT An aging human population has revealed the burden of chronic illness and age-related disease, and by understanding the genetic and environmental factors that drive aging, we will be better suited to develop and test therapeutics that slow age-related disease. As biological aging is influenced by both genetics and the environment, our laboratory studies the cellular and molecular drivers of aging, with a particular focus on inter- organelle communication in disease. Here, we newly describe a dramatic reorganization of endoplasmic reticulum (ER) subdomains in aging C. elegans. The ER mediates inter- and intracellular signaling through these sheet and tubule domains, and sheet:tubule balance is critical for cell function. ER tubules store calcium and lipids, and at specialized membrane contact sites, they regulate mitochondrial dynamics. We find that the aging ER undergoes a loss of rough ER sheets and expansion of smooth ER tubules, and our data suggest that modifying ER structure is sufficient to preserve mitochondrial morphology in age, making the ER a potential target in preventing age-related mitochondrial fragmentation. Though autophagy is seen as cytoprotective in aging, we show that autophagy is necessary for age-related ER remodeling. This may be explained by ER- phagy, a form of ER-selective autophagy that has not been studied in the context of aging, as ER-phagy shares common recycling processes. Finally, we demonstrate that caloric restriction, which extends lifespan, prevents this age-related loss of ER morphology. Therefore, we hypothesize that dysregulated ER-phagy drives age- related ER remodeling and that dietary restriction promotes longevity by mitigating this loss of ER form and function. To discern whether these changes are attributable to selective ER-phagy, rather than general autophagy, I will use a combination of in vivo imaging, fluorescent reporters, and RNAi to investigate the molecular mechanisms leading to a change in ER subdomains with age (Aim 1). In Aim 2, I will use dietary restriction, a robust longevity paradigm, to investigate the cause(s) and consequence(s) of ER remodeling in healthspan and lifespan regulation. This work will be conducted at Vanderbilt University under the supervision of Dr. Kristopher Burkewitz, Assistant Professor of Cell & Developmental Biology, who discovered roles for ER function in lifespan regulation through ER-mitochondrial crosstalk. I will additionally be supported by Dr. David Miller, Professor of Cell & Developmental Biology, whose lab is experienced in electron microscopy and pioneered many genetic engineering techniques I will perform in C. elegans. In these studies, I will receive feedback from a strong advisory committee with expertise including interorganelle signaling, membrane dynamics, and aging physiology. Successful completion of this project will not only advance our understanding of cell biology and the role of the ER in aging but also establish ER structure and function as therapeutic targets in the treatment of age-related disease.

项目概要/摘要 人口老龄化揭示了慢性疾病和与年龄相关的疾病的负担，并且 了解导致衰老的遗传和环境因素，我们将更适合开发和 测试减缓与年龄相关的疾病的疗法。由于生物衰老受到遗传和后天因素的影响 环境中，我们的实验室研究衰老的细胞和分子驱动因素，特别关注 疾病中的细胞器通讯。在这里，我们新描述了内质的戏剧性重组 衰老线虫中的网状结构 (ER) 子域。 ER 通过这些信号介导细胞间和细胞内信号传导 片层和小管结构域以及片层：小管平衡对于细胞功能至关重要。 ER 小管储存钙和 脂质，并在专门的膜接触位点，它们调节线粒体动力学。我们发现老化 ER 经历了粗糙 ER 表层的损失和光滑 ER 小管的扩张，我们的数据表明 改变 ER 结构足以在年龄上保持线粒体形态，使 ER 成为潜在的 预防与年龄相关的线粒体断裂的目标。尽管自噬被认为具有细胞保护作用 随着年龄的增长，我们发现自噬对于与年龄相关的内质网重塑是必要的。这可以用 ER 来解释： 自噬，一种 ER 选择性自噬形式，尚未在衰老背景下进行研究，如 ER-自噬一样 常见的回收流程。最后，我们证明，热量限制可以延长寿命，防止 这种与年龄相关的内质网形态丧失。因此，我们假设内质网自噬失调会导致年龄增长 相关的 ER 重塑以及饮食限制通过减轻 ER 形式的损失来促进长寿 功能。辨别这些变化是否归因于选择性 ER 吞噬，而不是一般性的 自噬，我将结合体内成像、荧光报告基因和 RNAi 来研究 导致 ER 亚结构域随年龄变化的分子机制（目标 1）。在目标 2 中，我将使用饮食 限制，一个强大的长寿范式，以研究 ER 重塑的原因和后果 健康寿命和寿命调节。这项工作将在范德比尔特大学的监督下进行 细胞与发育生物学助理教授 Kristopher Burkewitz 博士发现了 ER 的作用 通过 ER 线粒体串扰调节寿命。我还将得到大卫博士的支持 Miller，细胞与发育生物学教授，其实验室在电子显微镜和 开创了许多基因工程技术，我将在秀丽隐杆线虫中进行这些技术。在这些研究中，我将获得 来自强大的咨询委员会的反馈，其专业知识包括细胞器间信号传导、膜 动力学和衰老生理学。这个项目的成功完成不仅会增进我们的理解 细胞生物学和 ER 在衰老中的作用，同时也建立 ER 结构和功能作为治疗靶点 治疗与年龄相关的疾病。