Ex vivo rejuvenation and expansion of muscle stem cells from aged mice

衰老小鼠肌肉干细胞的离体再生和扩增

• 批准号: 8926619
• 负责人: Benjamin David Cosgrove
• 金额: $ 24.9万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8926619
• 关键词: Activities of Daily Living; Acute; Adhesions; Aging; Algorithms; Antibodies; Autologous; Biological Assay; Biology; Biomedical Engineering; Biomimetics; Cell Aging; Cell Culture Techniques; Cell physiology; Cellular Mechanotransduction; Cellular Stress; Clinical; Cytometry; Defect; Development; Faculty; Functional disorder; Generations; Homeostasis; Hydrogels; Individual; Injury; Institution; Laboratories; Least-Squares Analysis; Libraries; Life; Longevity; MAPK14 gene; Mentors; Mitogens; Modeling; Modification; Mus; Muscle; Muscle satellite cell; Muscular Atrophy; Natural regeneration; Pathway interactions; Performance; Phase; Phenotype; Phosphoproteins; Physiology; Population; Positioning Attribute; Protein Kinase; Quality of life; Recovery; Rejuvenation; Relative (related person); Research; Signal Transduction; Signaling Molecule; Skeletal Muscle; Stem cells; Surface; System; Systems Biology; Telomerase; Testing; Therapeutic; Tissues; Training; Transplant Recipients; Transplantation; Universities; Work; abstracting; adult stem cell; age related; aged; attenuation; base; career development; cell age; combinatorial; improved; in vivo; inhibitor/antagonist; injured; innovation; insight; mitogen-activated protein kinase p38; muscle form; muscle regeneration; muscle strength; non-invasive imaging; novel; programs; regenerative; research and development; response; satellite cell; senescence; small molecule; stem cell biology; stem cell fate; stem cell population; stem cell therapy; stemness; symposium; tissue regeneration; wasting

"Ex vivo Rejuvenation and Expansion of Muscle Stem Cells from Aged Mice" Project Summary/Abstract Muscle stem cells (MuSCs), also known as satellite cells, are essential to muscle regeneration throughout life [1]. In aging, skeletal muscle mass and regenerative capacity after injury progressively decline, leading to diminished quality of life in aged individuals [2]. Efforts to explain the dysfunction of aged skeletal muscle tissue have focused on aging-related changes in tissue microenvironment factors restricting MuSC function [3]. In my postdoctoral research, I have demonstrated, using non-invasive imaging assays of tissue regeneration, that MuSCs prospectively isolated from old mice have a marked reduction in regenerative capacity relative to young MuSCs, revealing a previously undetected intrinsic stem cell defect in old MuSCs. Further, I have identified a novel ex vivo strategy to overcome the regenerative dysfunction of old MuSCs; treatment of old MuSCs maintained on a soft biomimetic hydrogel platform [4] with a small molecule inhibitor of p38 mitogen- associated protein kinase yields an expansion in absolute numbers of functional stem cells and restores their function in regeneration to that of young MuSCs. This approach offers promise for rejuvenating and increasing the numbers of MuSCs and could enable localized autologous stem cell therapy for muscle wasting in aged individuals, for which there are no pharmacologic treatments in clinical use. I propose to merge prior training in bioengineering and stem cell biology with new training in muscle physiology and systems biology to further investigate the regenerative dysfunction of MuSCs in aging and its rescue ex vivo. In Aim 1 (K99 phase), I will evaluate whether ex vivo-treated old MuSCs can rescue defective muscle regeneration and increase muscle strength in old mice and are capable of long-term rejuvenated function in response to successive regenerative demands. In Aim 2 (bridging K99/R00 phases), I will elucidate whether defective regenerative function is a homogeneous or heterogeneous phenotype in old MuSCs by combining multi-parameter mass cytometry (CyTOF) [5] and SPADE algorithm [6] analysis with sensitive transplantation assays to identify and compare the regenerative functions of MuSC sub-populations isolated from young and old mice. In Aim 3 (R00 phase), I will elucidate dysregulated signaling network mechanisms underlying the stem cell dysfunction of old MuSCs for improved therapeutic treatment using signaling network- level systems biology approaches [7]. This Transition to Independence proposal describes research and career development activities, including conference attendance and course training, that will establish me as a competitive candidate for an independent faculty position and aid my development of an innovative, successful research program in the biology and treatment of stem cell aging. These activities will be mentored by Drs. Helen Blau (primary), Scott Delp (co-mentor), and Garry Nolan (co-mentor) at Stanford University, which is a world-class stem cell biology research institution.

“从老年小鼠的体内复兴和肌肉干细胞的扩张” 项目摘要/摘要 肌肉干细胞（MUSC），也称为卫星细胞，对于一生都至关重要 [1]。在衰老中，受伤后骨骼肌质量和再生能力逐渐下降，导致 老年人的生活质量下降[2]。努力解释老年骨骼肌的功能障碍 组织的重点是与衰老相关的组织微环境因子限制MUSC功能的变化[3]。 在我的博士后研究中，我已经使用非侵入性成像测定组织进行了研究， 从旧小鼠中前瞻性隔离的MUSC相对于再生能力显着降低 年轻的MUSC揭示了旧MUSC中先前未发现的固有干细胞缺陷。此外，我有 确定了一种新型的离体策略，以克服旧MUSC的再生功能障碍。旧的处理 MUSC维持在柔软的仿生水凝胶平台上[4]，该平台具有p38的小分子抑制剂 相关蛋白激酶会在绝对数量的功能干细胞中扩展并恢复其 在年轻MUSC的再生中起作用。这种方法为恢复和增加提供了希望 MUSC的数量，可以使局部自体干细胞疗法用于肌肉浪费 个人，在临床用途中没有药理学治疗。 我建议将生物工程和干细胞生物学的事先培训与肌肉的新培训合并 生理和系统生物学，以进一步研究MUSC在衰老及其ITS中的再生功能障碍 救援前体内。在AIM 1（K99阶段）中，我将评估过体内处理的旧MUSC是否可以营救有缺陷的MUSC 肌肉再生并增加旧小鼠的肌肉力量，并能够长期恢复活力 响应连续再生需求的功能。在AIM 2（桥接K99/R00阶段）中，我将阐明 在旧MUSC中，有缺陷的再生功能是均质还是异质表型 将多参数质量细胞仪（Cytof）[5]和Spade算法[6]结合在一起 移植测定法以识别和比较分离的MUSC亚群的再生功能 来自老鼠和老鼠。在AIM 3（R00阶段）中，我将阐明失调的信号网络机制 使用信号网络的旧MUSC的干细胞功能障碍以改善治疗治疗的基础 水平系统生物学方法[7]。 向独立建议的过渡描述了研究和职业发展活动， 包括会议出勤和课程培训，这将使我成为一个竞争性的候选人 独立的教师职位，并帮助我制定创新，成功的研究计划 干细胞衰老的生物学和治疗。这些活动将由博士指导。海伦·布劳（Helen Blau）（主要），斯科特 斯坦福大学的Delp（Co-Incortor）和Garry Nolan（Co-​​Centor），这是世界一流的干细胞生物学 研究机构。