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: 10600362
• 负责人: James P. White
• 金额: $ 12.32万
• 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/10600362
• 关键词: Aging; Apoptosis; Autophagocytosis; Award; Biological Assay; Biology of Aging; Cell Aging; Cell Cycle; Cell Separation; Cell Survival; Cellular Metabolic Process; Cellular biology; Clinical; Data; Development; Development Plans; Educational workshop; Elderly; Ensure; Environment; Exercise; Human; Impairment; Individual; Injury; Learning; Mentors; Mentorship; Metabolic; Metabolic dysfunction; Methods; Molecular; Morbidity - disease rate; Mus; Muscle; Muscle satellite cell; Muscular Atrophy; Natural regeneration; Organism; Pathway interactions; Persons; Phenotype; Physical Function; Physiological; Predisposition; Proliferating; Quality of life; Regenerative capacity; Research Activity; Research Personnel; Role; Skeletal Muscle; Testing; Training; Training Activity; Training Programs; Transplantation; Wages; Work; age related; aged; career; career development; cell age; comorbidity; disability; experimental study; functional loss; human model; improved; in vivo; injured; injury and repair; medical schools; mouse model; muscle form; muscle regeneration; muscle strength; prevent; professor; research and development; sarcopenia; satellite cell; 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. 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- p27Kip pathway appears critical for successful transition from quiescence to entry into the cell cycle. Our preliminary data identify perturbations in the AMPK-p27Kip pathway with advanced age. This award period will investigate the role of the AMPK-p27Kip 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 target p27Kip regulates 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-p27Kip pathway, thereby enhancing proliferation and metabolic function in aging murine and human satellite cells. In Aim 3, we will test the hypothesis that rescuing activity of the AMPK-p27Kip pathway in aged satellite cells will improve in vivo regenerative capacity by transplantation experiments in injured muscle of young and old mice. Together, the experiments in this proposal will test the hypothesis that the AMPK- p27Kip pathway is impaired in aging satellite cells; and the hypothesis that this impairment contributes to the delayed proliferation rate, susceptibility to apoptosis, and reduced ability to regenerate muscle after injury. 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. Dr. White will also have Drs. Bruce Spiegelman, Amy Wagers and Ana Maria Cuervo as co-mentors; they will respectively enhance training in cell metabolism; aging stem cell biology; and autophagy. The Duke School of Medicine environment 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 包括身体机能的丧失肌肉减少症患者的一个这样的扰动是降低的能力， 使受伤后的肌肉再生肌肉干细胞，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。衰老的卫星细胞 在损伤时激活;易受细胞凋亡影响;并且在修复受损肌肉中效率较低。AMPK- p27 Kip通路对于从静止到进入细胞周期的成功过渡似乎是关键的。我们 初步数据证实了AMPK-p27 Kip通路随年龄增长而紊乱。本次颁奖期间 将研究AMPK-p27 Kip通路在人类和哺乳动物卫星细胞衰老表型中的作用。 小鼠模型。在Aim 1中，我们将检验AMPK及其下游靶点p27 Kip的激活是否与AMPK的激活有关的假设。 调节衰老卫星细胞中的自噬/凋亡决定。我们将使用分子检测来拯救 衰老细胞中该途径的功能丧失并恢复增殖能力。在目标2中，我们将测试 假设运动是AMPK和自噬的生理诱导剂，刺激AMPK-p27 Kip 途径，从而增强衰老小鼠和人类卫星细胞的增殖和代谢功能。在 目标3，我们将检验以下假设：拯救衰老卫星细胞中AMPK-p27 Kip途径的活性将 通过在年轻和老年小鼠损伤肌肉中移植实验来提高体内再生能力。 总之，本研究中的实验将检验AMPK-p27 Kip通路受损的假设。 在老化的卫星细胞中;以及这种损伤有助于延迟增殖率的假设， 细胞凋亡的易感性和损伤后再生肌肉的能力降低。白色博士的主要方面 职业发展将是：学习如何协调临床运动试验;培训卫星方法 细胞分离和代谢分析，特别是在衰老生物体的背景下。培训计划将 需要专门的内部和外部科学演示;干细胞相关课程/研讨会 生物学和老龄化;和密集的职业导师，以确保走向独立的进展。研究 和职业发展计划，将与一个优秀的导师团队进行。 博士William E.克劳斯是杜克医学院的教授，是临床锻炼方面的专家 研究和肌肉/卫星细胞生物学;他将担任主要导师。白色博士也将有博士。 布鲁斯Spiegelman，艾米赌注和安娜玛丽亚Cuervo作为共同导师，他们将分别提高 细胞代谢训练;衰老干细胞生物学;和自噬。杜克医学院 环境是开展本提案所述研究和培训活动的理想环境。该奖项将提供 博士白色与最佳的培训，以确保一个出色的开始，他的职业生涯作为一个独立的调查员。