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. 项目概要/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，它会导致许多并发症； 包括身体机能丧失在内的肌少症患者的此类困扰之一是能力下降。 受伤后再生肌肉需要激活肌肉干细胞，称为卫星细胞。 老化的卫星细胞增殖和分化以再生肌肉和恢复身体功能的速度较慢。 损伤时激活；容易发生细胞凋亡；修复受损肌肉的效率较低。 p27Kip 通路对于从静止状态成功过渡到进入细胞周期至关重要。 初步数据表明 AMPK-p27Kip 通路中的扰动与高龄有关。 将研究 AMPK-p27Kip 通路在人类和卫星细胞衰老表型中的作用 在 Aim1 小鼠模型中，我们将测试 AMPK 及其下游靶点 p27Kip 激活的假设。 调节老化卫星细胞的自噬/凋亡决定，我们将使用分子测定来拯救。 在目标 2 中，我们将测试衰老细胞中该通路的功能丧失和增殖能力。 假设运动是 AMPK 和自噬的生理诱导剂，可刺激 AMPK-p27Kip 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。 目标 3，我们将测试以下假设：老化卫星细胞中 AMPK-p27Kip 通路的拯救活性将 通过年轻和年老小鼠受伤肌肉的移植实验提高体内再生能力。 总之，本提案中的实验将检验 AMPK-p27Kip 通路受损的假设 在老化的卫星细胞中；以及这种损伤导致增殖速度延迟的假设， 细胞凋亡的易感性以及受伤后肌肉再生能力的降低是怀特博士的关键方面。 职业提升将是：学习如何协调临床运动试验，并进行卫星方法培训； 细胞分离和代谢分析，特别是在衰老有机体的背景下。 需要专门的内部和外部干细胞相关课程/研讨会； 生物学和衰老；以及密集的职业指导，以确保研究的独立性。 本提案中详细的职业发展计划将由优秀的导师团队进行。 杜克医学院教授 William E. Kraus 博士是临床锻炼方面的专家 研究和肌肉/卫星细胞生物学；他将担任怀特博士的主要导师。 Bruce Spiegelman、Amy Wagers 和 Ana Maria Cuervo 将分别担任联合导师； 细胞代谢、衰老干细胞生物学和自噬培训。 该奖项将提供适合本提案中概述的研究和培训活动的环境。 怀特博士接受过最佳培训，以确保他作为独立调查员的职业生涯有一个良好的开端。

项目成果

The AMPK/p27Kip1 Axis Regulates Autophagy/Apoptosis Decisions in Aged Skeletal Muscle Stem Cells.

AMPK/p27Kip1 轴调节衰老骨骼肌干细胞的自噬/凋亡决策。

• 发表时间: 2018-08-14
• 影响因子: 5.9
• 作者: White, James P;Billin, Andrew N;Campbell, Milton E;Russell, Alan J;Huffman, Kim M;Kraus, William E
• 通讯作者: Kraus, William E

Autophagy as a Therapeutic Target to Enhance Aged Muscle Regeneration.

自噬作为增强衰老肌肉再生的治疗靶点。

• 发表时间: 2019
• 影响因子: 6
• 作者: Lee, David E;Bareja, Akshay;Bartlett, David B;White, James P
• 通讯作者: White, James P

Maximizing Longevity and Healthspan: Multiple Approaches All Converging on Autophagy.

最大限度地延长寿命和健康寿命：多种方法都集中在自噬上。

• 发表时间: 2019
• 作者: Bareja, Akshay;Lee, David E;White, James P
• 通讯作者: White, James P

The AMPK/p27Kip1 Pathway as a Novel Target to Promote Autophagy and Resilience in Aged Cells.

AMPK/p27Kip1 通路作为促进衰老细胞自噬和恢复能力的新靶点。

• 发表时间: 2021
• 影响因子: 6
• 作者: McKay, Lauren K;White, James P
• 通讯作者: White, James P