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

基本信息

批准号：
8515285
负责人：
Benjamin David Cosgrove
金额：
$ 8.83万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8515285
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 Image 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 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 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

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：肌肉干细胞（MUSC），也称为卫星细胞，对肌肉再生至关重要[1]。在衰老中，骨骼肌质量和受伤后的再生能力逐渐下降，导致生活质量降低在老年人中[2]。解释老年骨骼肌组织功能障碍的努力集中在与衰老相关的微环境因子限制MUSC功能的变化[3]。在我的博士后研究中，我证明，使用非侵入性成像测定组织再生，从旧小鼠中前瞻性隔离的MUSC相对于年轻的MUSC，再生能力显着降低，揭示了先前未发现的旧MUSC中未发现的固有干细胞缺陷。此外，我已经确定了一种新型的体内策略，以克服旧MUSC的再生功能障碍。用小的p38促丝裂料相关蛋白激酶的小分子抑制剂维持在软仿生水凝胶平台上的旧MUSC的处理可产生绝对数量的功能干细胞，并恢复其功能，以重新恢复到年轻MUSC的功能。这种方法为恢复和增加MUSC的数量提供了希望，并可以使年龄个体的肌肉浪费局部自体干细胞疗法，为此，在临床用途中没有药理治疗。我建议将生物工程和干细胞生物学的事先培训与肌肉生理和系统生物学领域的新培训合并，以进一步研究MUSC在衰老中的再生功能障碍及其救援事件。在AIM 1（K99阶段）中，我将评估经体治疗的旧MUSC是否可以挽救有缺陷的肌肉再生并增加旧小鼠的肌肉力量，并且能够响应连续的再生需求而具有长期的复兴功能。在AIM 2（桥接K99/R00阶段）中，我将通过将多参数质量细胞术（Cytof）[5]和Spade算法分析与对识别的敏感移植功能相结合，通过将多参数质量细胞术（Cytof）结合到旧MUSC中是一种同质或异构表型，是一种同质或异质表型。老鼠。在AIM 3（R00相）中，我将阐明使用信号网络水平系统生物学方法改善旧MUSC的干细胞功能障碍的信号网络机制[7]。向独立提案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，这将使我成为独立教师职位的竞争候选人，并帮助我开发创新的，成功的研究研究计划，以实现干细胞衰老的生物学和治疗。这些活动将由博士指导。斯坦福大学的Helen Blau（小学），Scott Delp（Co-Contor）和Garry Nolan（Co-Contor），这是世界一流的干细胞生物学研究机构。