Molecular Mechanisms Regulating Motility and Migration of Muscle Satellite Cells

调节肌肉卫星细胞运动和迁移的分子机制

• 批准号: 7907839
• 负责人: Dawn D Cornelison
• 金额: $ 19.16万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7907839
• 关键词: 3-Dimensional; Acute; Address; Adhesions; Adult; Affect; Antibodies; Area; Basic Science; Behavior; Binding; Biological; Biological Assay; Blocking Antibodies; Cell Adhesion; Cell Communication; Cell Culture Techniques; Cell Surface Proteins; Cell Therapy; Cell physiology; Cells; Clinical; Collaborations; Collagen; Cues; Cytoskeleton; DNA Sequence Rearrangement; Data; Defect; Development; Disease; Embryo; Endothelial Cells; Engraftment; Environment; Event; Exhibits; Explosion; Extracellular Matrix; Fiber; Fibroblast Growth Factor 2; Fibroblasts; Foundations; Goals; Grant; Growth Factor; Hepatocyte Growth Factor; Heterogeneity; Home environment; In Vitro; Individual; Infection; Injection of therapeutic agent; Injury; Integrins; Intrinsic factor; Ligands; Lipids; Measures; Mediating; Methods; Modeling; Molecular; Monomeric GTP-Binding Proteins; Mus; Muscle; Muscle Fibers; Muscle satellite cell; Muscular Dystrophies; Mutagens; Mutant Strains Mice; Mutation; Myoblasts; Natural regeneration; Neurons; Peptides; Pharmacologic Substance; Population; Procedures; Proliferating; Protein Isoforms; Proteins; Protocols documentation; Receptor Signaling; Regulation; Research Personnel; Role; Signal Pathway; Signal Transduction; Site; Skeletal Muscle; Stem cells; Stimulus; Surveys; System; Techniques; Time; Tissues; Vertebrates; Video Microscopy; Viral; Viral Vector; Work; adult stem cell; base; cell motility; chemokine; extracellular; gene therapy; in vivo; inhibitor/antagonist; insight; interest; migration; mimetics; muscle regeneration; mutant; myogenesis; novel; population migration; public health relevance; receptor; repaired; response; rho; satellite cell; syndecan-4; therapeutic effectiveness; tool; vector

DESCRIPTION (provided by applicant): Repair and replacement of damaged skeletal muscle in vertebrates requires the activity of a population of resident adult stem cells (satellite cells.) Following acute injury, these relatively rare and highly dispersed cells are required to respond by quickly becoming `activated', multiplying to produce a large pool of replacement myoblasts, relocating to the site of the injury, and differentiating to form new muscle fibers. The molecular and cellular processes required for adult myogenesis (regeneration) overlap with, but are largely distinct from, those active in the embryo when muscle is first formed. At the level of the cell's interactions with the extracellular environment, the requirements for satellite cell motility and migration within the tissue are a critical aspect of muscle regeneration that has not been well explored. In collaboration with Dr. George Davis, we have combined primary satellite cell culture on their host myofibers in a programmable 3D collagen matrix with timelapse videomicroscopy to develop a novel system for qualitatively and quantitatively examining satellite cell migration on their native substrate. Conditions can be altered by addition of exogenous stimuli such as potential mutagens, blocking of soluble or cell-surface proteins with antibodies or peptide mimetics, addition of pharmacological inhibitors of specific signaling pathways or cytoskeletal remodelers, specific infection of satellite cells with viral expression constructs, or use of fibers and satellite cells derived from targeted mutations; all of these can also be assayed both individually and in combination. We have used this system to assess the roles of soluble factors such as growth factors, chemokines and signaling lipids; extracellular matrix and adhesion factors; transmembrane signaling receptors and integrins; and intracellular modulators of the cytoskeleton. Our goal for this short-term exploratory grant is to extend and refine these results to build a working model of satellite cell migration in the context of these defined classes of signaling factors and effectors. Once we have identified critical interactions and points of control, we will continue on to ask how such activities are integrated within individual satellite cells to effect a coherent migration of the population of proliferating myoblasts towards an area of injury. Our broad, long-term goal is to understand how satellite cells detect, integrate, and respond appropriately in time and space to the transient and dynamic signaling environment that would constitute a muscle injury in vivo, with the biological robustness that is taken for granted in healthy muscle tissue and compromised in dystrophic muscle. PUBLIC HEALTH RELEVANCE: In addition to addressing an underexplored facet of basic research into the mechanisms of adult myogenesis, this project has high potential to contribute to the development of satellite cell- based therapies for diseases such as the muscular dystrophies. A critical area of concern in current adult myoblast and muscle-derived stem cell engraftment procedures is the unmet requirement for injected cells to, at least, disperse broadly from the injection site or, at best, actively home to either investigator-defined sites or sites of maximum damage. By providing insight into the motogenic stimuli, preferred migration substrate, and specific guidance cues used by satellite cells migrating in a system (single fiber culture) that could be expected to recapitulate many of the influences found in vivo, this work will ideally suggest potential avenues to modify current myoblast engraftment protocols to enhance their therapeutic effectiveness.

描述（由申请人提供）：脊椎动物受损骨骼肌的修复和替换需要一群常驻成体干细胞（卫星细胞）的活性。急性损伤后，需要这些相对稀少且高度分散的细胞通过快速“激活”、增殖以产生大量替代成肌细胞、重新定位至损伤部位并分化以形成新的肌纤维来作出反应。成人肌肉发生（再生）所需的分子和细胞过程与肌肉首次形成时胚胎中的活性过程重叠，但在很大程度上不同。在细胞与细胞外环境相互作用的水平上，组织内卫星细胞运动和迁移的要求是肌肉再生的一个关键方面，尚未得到很好的探索。在与博士合作乔治戴维斯，我们结合了原代卫星细胞培养在其主机肌纤维在可编程的三维胶原蛋白基质与延时视频显微镜开发一种新的系统，定性和定量检查卫星细胞迁移其原生基板。可以通过加入外源性刺激如潜在诱变剂、用抗体或肽模拟物阻断可溶性或细胞表面蛋白、加入特异性信号传导途径或细胞骨架重塑剂的药理学抑制剂、用病毒表达构建体特异性感染卫星细胞或使用源自靶向突变的纤维和卫星细胞来改变条件;所有这些也可以单独或组合进行测定。我们已经使用这个系统来评估可溶性因子的作用，如生长因子，趋化因子和信号脂质;细胞外基质和粘附因子;跨膜信号受体和整合素;和细胞骨架的细胞内调节剂。我们这项短期探索性资助的目标是扩展和完善这些结果，以在这些定义的信号传导因子和效应物类别的背景下建立卫星细胞迁移的工作模型。一旦我们确定了关键的相互作用和控制点，我们将继续询问这些活动是如何整合到单个卫星细胞中的，以实现增殖的成肌细胞群体向损伤区域的一致迁移。我们广泛的，长期的目标是了解卫星细胞如何检测，整合，并在时间和空间上适当地响应瞬态和动态信号环境，这将构成体内肌肉损伤，具有在健康肌肉组织中被认为是理所当然的生物稳健性，并在营养不良的肌肉中受到损害。 公共卫生相关性：除了解决成人肌生成机制基础研究中一个未充分探索的方面外，该项目还具有很大的潜力，有助于开发基于卫星细胞的治疗方法，用于肌营养不良症等疾病。在目前的成人成肌细胞和肌肉来源的干细胞移植程序中，一个关键的关注领域是对注射细胞的未满足的要求，即至少从注射部位广泛分散，或者最好主动归巢到植入器定义的部位或最大损伤的部位。通过提供深入了解运动刺激，首选的迁移基板，和特定的指导线索所使用的卫星细胞迁移在一个系统（单纤维培养），可以预期，以概括许多在体内发现的影响，这项工作将理想地建议潜在的途径来修改目前的成肌细胞移植协议，以提高其治疗效果。

项目成果

Ephrin-A3 promotes and maintains slow muscle fiber identity during postnatal development and reinnervation.

在产后发育和加剧期间，ephrin-A3在产后发育和加重过程中促进和维持较慢的肌肉纤维身份。

• DOI: 10.1083/jcb.201502036
• 发表时间: 2015-12-07
• 期刊: The Journal of cell biology
• 作者: Stark DA;Coffey NJ;Pancoast HR;Arnold LL;Walker JP;Vallée J;Robitaille R;Garcia ML;Cornelison DD
• 通讯作者: Cornelison DD

3D timelapse analysis of muscle satellite cell motility.

• DOI: 10.1002/stem.178
• 发表时间: 2009-10
• 期刊: STEM CELLS
• 影响因子: 5.2
• 作者: Siegel, Ashley L.;Atchison, Kevin;Fisher, Kevin E.;Davis, George E.;Cornelison, D. D. W.
• 通讯作者: Cornelison, D. D. W.

Muscle satellite cell proliferation and association: new insights from myofiber time-lapse imaging.

• DOI: 10.1186/2044-5040-1-7
• 发表时间: 2011-02-02
• 期刊: Skeletal muscle
• 影响因子: 4.9
• 作者: Siegel AL;Kuhlmann PK;Cornelison DD
• 通讯作者: Cornelison DD

CD34 promotes satellite cell motility and entry into proliferation to facilitate efficient skeletal muscle regeneration.

• DOI: 10.1002/stem.759
• 发表时间: 2011-12
• 期刊: STEM CELLS
• 影响因子: 5.2
• 作者: Alfaro, Leslie Ann So;Dick, Sarah A.;Siegel, Ashley L.;Anonuevo, Adam S.;McNagny, Kelly M.;Megeney, Lynn A.;Cornelison, D. D. W.;Rossi, Fabio M. V.
• 通讯作者: Rossi, Fabio M. V.