The AMPK/ULK1/p27Kip1 axis regulates autophagy and cell survival in aged satellite cells

AMPK/ULK1/p27Kip1 轴调节衰老卫星细胞的自噬和细胞存活

• 批准号: 10177828
• 负责人: James P. White
• 金额: $ 12.25万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10177828
• 关键词: 5' -AMP-activated protein kinase; Acute; Address; Affect; Age-Years; Aging; Apoptosis; Apoptotic; Autophagocytosis; Award; Biological Assay; Biology of Aging; Caloric Restriction; Cell Aging; Cell Cycle; Cell Proliferation; Cell Separation; Cell Survival; Cell physiology; Cellular Metabolic Process; Cellular biology; Chronic; Clinical; Cytosol; Data; Development; Development Plans; Educational workshop; Elderly; Ensure; Environment; Equilibrium; Exercise; Functional disorder; Goals; Health; Health Care Costs; Healthcare; Human; Impairment; Individual; Injury; Lead; Learning; Mediating; Mentors; Mentorship; Metabolic; Metabolic dysfunction; Metabolism; Methods; Molecular; Molecular Target; Morbidity - disease rate; Mus; Muscle; Muscle satellite cell; Muscular Atrophy; Natural regeneration; Organism; Outcome; Pathway interactions; Persons; Pharmacology; Phenotype; Phosphorylation; Physical Function; Physiological; Play; Predisposition; Process; Proliferating; Protein Kinase; Quality of life; Regenerative capacity; Research Activity; Research Personnel; Research Training; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Testing; Time; Training; Training Activity; Training Programs; Wages; Work; age related; age-related muscle loss; aged; cancer cell; care burden; career; career development; cell age; comorbidity; disability; effective therapy; endurance exercise; experimental study; frailty; functional loss; human model; improved; injury and repair; loss of function; medical schools; mortality; mouse model; muscle form; muscle regeneration; muscle strength; novel; preservation; prevent; professor; reduced muscle strength; research and development; resilience; restoration; sarcopenia; satellite cell; skeletal preservation; stem cell aging; stem cell biology; stem cell function

a. Project summary/abstract: Sarcopenia is the age-related loss in skeletal muscle mass and strength; it leads to a host of co-morbidities including loss of physical function and overall resilience. One such perturbation in persons with sarcopenia is the diminished ability to regenerate muscle after injury. Muscle stem cells, referred to as satellite cells, are required to activate, proliferate and differentiate to regenerate muscle and restore physical function. Aged satellite cells are slower to activate upon injury; susceptible to apoptosis; and less efficient in repairing injured muscle. The AMPK/ULK1/p27Kip1 pathway appears critical for successful transition from quiescence to entry into the cell cycle. Our preliminary data identify perturbations in the AMPK/ULK1/p27Kip1 pathway with advanced age. This award period will investigate the role of the AMPK/ULK1/p27Kip1 pathway in the phenotype of satellite cell aging in both human and mouse models. In Aim1, we will test the hypothesis that activation of AMPK and its downstream targets ULK1 and p27Kip1 regulate the autophagy/apoptosis decision in aged satellite cells. We will use molecular assays to rescue the functional loss of this pathway in aged cells and return proliferative capacity. In Aim 2, we will test the hypothesis that exercise, a physiological inducer of AMPK and autophagy, stimulates the AMPK/ULK1/p27Kip1 pathway, thereby enhancing proliferation and metabolic function in aging murine and human satellite cells. Aim 3 will test the hypothesis that AMPK/ULK1/p27Kip1 signaling will regulate the beneficial effects of caloric restriction on aged satellite cells. Together, the experiments in this proposal will test the hypothesis that the AMPK/ULK1/p27Kip1 pathway is impaired in aging satellite cells resulting in a reduction in autophagy and susceptibility to apoptosis. Key aspects of Dr. White's career enhancement will be: to learn how to coordinate clinical exercise trials; to train in methods of satellite cell isolation and metabolic analysis, especially in the context of the aging organism. The training program will entail dedicated internal and external scientific presentations; pertinent course work/workshops in stem cell biology and aging; and intensive career mentorship to ensure progress toward independence. The research and career development plan detailed in this proposal will be conducted with a team of outstanding mentors. Dr. William E. Kraus, a professor at the Duke Medical School is an established expert in clinical exercise studies and muscle/satellite cell biology; he will serve as the primary mentor. Drs. Kenneth Schmader, Deborah Muoio (Duke) and Amy Wagers (Harvard) will serve as co-mentors; they will facilitate training in aging biology, cell metabolism and aging stem cell biology, respectively. The environment at the Duke School of Medicine is ideal for the research and training activities outlined in this proposal. This award will provide Dr. White with optimal training to ensure an outstanding start to his career as an independent investigator.

一个。项目概要/摘要： 肌肉减少症是指与年龄相关的骨骼肌质量和力量损失；它会导致一系列并发症 包括身体机能和整体恢复能力的丧失。肌肉减少症患者的此类困扰之一是 受伤后肌肉再生能力下降。肌肉干细胞，称为卫星细胞，是 激活、增殖和分化以再生肌肉和恢复身体功能所需。老化 卫星细胞受伤后激活速度较慢；易发生细胞凋亡；且修复受伤的效率较低 肌肉。 AMPK/ULK1/p27Kip1 通路对于从静止状态到进入状态的成功转变至关重要 进入细胞周期。我们的初步数据确定了 AMPK/ULK1/p27Kip1 通路中的扰动 高龄。该奖励期将研究 AMPK/ULK1/p27Kip1 通路在 人类和小鼠模型中卫星细胞衰老的表型。在目标 1 中，我们将检验以下假设： AMPK 及其下游靶标 ULK1 和 p27Kip1 的激活调节自噬/凋亡决策 在老化的卫星细胞中。我们将使用分子检测来挽救衰老细胞中该通路的功能丧失 并恢复增殖能力。在目标 2 中，我们将检验以下假设：运动是一种生理诱导剂 AMPK 和自噬，刺激 AMPK/ULK1/p27Kip1 通路，从而增强增殖和 衰老小鼠和人类卫星细胞的代谢功能。目标 3 将检验以下假设： AMPK/ULK1/p27Kip1 信号传导将调节热量限制对老化卫星细胞的有益影响。 总之，本提案中的实验将检验 AMPK/ULK1/p27Kip1 途径的假设 老化卫星细胞受损，导致自噬减少和细胞凋亡敏感性降低。钥匙 怀特博士职业提升的方面将是： 学习如何协调临床运动试验；训练 卫星细胞分离和代谢分析方法，特别是在衰老有机体的背景下。 培训计划将包括专门的内部和外部科学演示；相关课程 干细胞生物学和衰老工作/研讨会；和密集的职业指导，以确保取得进展 独立。本提案中详述的研究和职业发展计划将与 优秀的导师团队。杜克大学医学院教授 William E. Kraus 博士是一位知名人士 临床运动研究和肌肉/卫星细胞生物学专家；他将担任主要导师。博士。 Kenneth Schmader、Deborah Muoio（杜克大学）和 Amy Wagers（哈佛大学）将担任联合导师；他们会 分别促进衰老生物学、细胞代谢和衰老干细胞生物学的培训。环境 杜克大学医学院非常适合本提案中概述的研究和培训活动。这 该奖项将为怀特博士提供最佳培训，以确保他作为一名医生的职业生涯有一个良好的开端 独立调查员。