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.

a.项目总结/摘要： 肌肉减少症是与年龄相关的骨骼肌质量和力量的损失;它导致了一系列的合并症 包括身体功能和整体恢复力的丧失。在肌肉减少症患者中的一种这样的扰动是 受伤后肌肉再生能力减弱。肌肉干细胞，被称为卫星细胞， 需要激活，增殖和分化，以再生肌肉和恢复身体功能。岁 卫星细胞在损伤后激活较慢;易于凋亡;并且在修复损伤中效率较低 肌肉. AMPK/ULK 1/p27 Kip 1通路对于从静止到进入的成功转变似乎是关键的 进入细胞周期。我们的初步数据确定了AMPK/ULK 1/p27 Kip 1通路的扰动， 高龄该奖项期间将研究AMPK/ULK 1/p27 Kip 1通路在细胞凋亡中的作用。 在人类和小鼠模型中的卫星细胞衰老表型。在Aim 1中，我们将测试以下假设： AMPK及其下游靶点ULK 1和p27 Kip 1的激活调节自噬/凋亡决定 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这一通路的功能丧失 恢复增殖能力。在目标2中，我们将检验这样一个假设，即运动是一种生理诱导剂， AMPK和自噬，刺激AMPK/ULK 1/p27 Kip 1通路，从而增强增殖， 衰老的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/ULK 1/p27 Kip 1信号通路将调节热量限制对衰老卫星细胞的有益作用。 总之，本提案中的实验将检验AMPK/ULK 1/p27 Kip 1通路是 在老化的卫星细胞中受损，导致自噬减少和对凋亡的敏感性。关键 白色的职业发展方面将是：学习如何协调临床运动试验;培训 在卫星细胞分离和代谢分析方法中，特别是在衰老生物体的背景下。 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方面的工作/研讨会;以及密集的职业指导，以确保 独立本建议书中详细说明的研究和职业发展计划将通过以下方式进行： 优秀的导师团队。William E.克劳斯是杜克医学院的教授， 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。Drs. Kenneth Schmader，Deborah Muoio（杜克）和Amy Wagers（哈佛）将担任共同导师;他们将 分别促进老化生物学、细胞代谢和老化干细胞生物学的培训。环境 在杜克医学院是理想的研究和培训活动中概述了这一建议。这 该奖项将为白色博士提供最佳培训，以确保他的职业生涯有一个出色的开端， 独立调查员