Molecular mechanisms of muscle stem cells transitioning intoquiescence

肌肉干细胞转变为静止状态的分子机制

• 批准号: 8715431
• 负责人: Christoph Lepper
• 金额: $ 39万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-20 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8715431
• 关键词: ARHGEF5 gene; Accidents; Acute; Adult; Alleles; Automobile Driving; Behavior; Bioinformatics; Biological Assay; Candidate Disease Gene; Cell Adhesion; Cell Death; Cell Polarity; Cell Proliferation; Cell Separation; Cell Therapy; Cells; Characteristics; Chimeric Proteins; Clinical; Cytoskeleton; Data; Defect; Development; Epigenetic Process; Essential Genes; Estrogen Receptors; Future; Gene Expression Profile; Genes; Genetic; Genetic Programming; Injury; Intramuscular; Knock-in Mouse; Life; Locomotion; Longitudinal Studies; Mechanics; Mediating; Messenger RNA; Modeling; Molecular; Movement; Mus; Muscle; Muscle satellite cell; Muscular Dystrophies; Natural regeneration; Newborn Infant; Pathway interactions; Play; Population; Posture; Process; Production; Proliferating; Property; Proteins; Regulation; Reporter; Role; Signal Transduction; Skeletal Muscle; Sorting - Cell Movement; Source; Staging; Stem cells; Tamoxifen; Testing; Time; Transplantation; Viral; base; combat; comparative; critical period; gene function; knock-down; mouse model; muscle regeneration; mutant; novel; programs; recombinase; repaired; satellite cell; screening; small hairpin RNA; stem cell population; transcription factor

PROJECT SUMMARY The mammalian musculature provides mechanical force for locomotion. Muscles display remarkable ability to regenerate from reservoirs of muscle stem cells. How muscle stem cells transition from an actively proliferating population during development into a reserved population in adulthood for muscle repair/regeneration throughout lifetime is not known. I have obtained evidence that, in the mouse model, this process depends on the function of the Pax7 gene, which encodes a transcription factor. I plan the following aims to define 1) the precise role of Pax7, 2) the transcriptome changes during the development-to-adult transition, and 3) other essential genes (acting in parallel or downstream of Pax7) that mediate this transition. Aim 1: Determining the cellular and molecular defects of Pax7 mutant satellite cells. When and how do Pax7 mutant cells become defective in the development-to-adult transition? I will combine a plethora of assays for determining several muscle stem cell properties, including muscle differentiation, cell proliferation, cell death, niche occupancy, and cell polarity. These assays will be conducted with time course studies from newborn to adult. Aim 2: Performing mRNA-seq of Pax7 control and Pax7 mutant satellite cells. What are the transcriptome changes during this transition and what are the specific processes governed by Pax7? A comparative time course study of transcriptomes between control and Pax7 mutant cells throughout the development-to-adult transitional period will be performed using mRNA-seq, followed by bioinformatic analysis. Aim 3: Screening candidate genes for their function in satellite cells. What are the genes (in addition to Pax7) critical for the development-to-adult muscle stem cell transition? From Aim 2, I will select up to 50 genes to test for their function in this aspect; they may either act independently or downstream of Pax7. I will develop a novel assay for rapid screening of these genes' functions by combining gene knock-down and intramuscular transplantation of muscle stem cells.

项目概要 哺乳动物的肌肉组织为运动提供机械力。肌肉显示出非凡的能力 从肌肉干细胞库中再生。肌肉干细胞如何从活跃增殖的状态转变为 发育成成年后肌肉修复/再生保留群体的人口 整个一生不得而知。我获得的证据表明，在小鼠模型中，这个过程取决于 Pax7 基因的功能，该基因编码转录因子。我计划以下目标来定义 1） Pax7 的精确作用，2) 转录组在发育到成人过渡期间的变化，以及 3) 其他 介导这种转变的必需基因（与 Pax7 平行或下游起作用）。 目标 1：确定 Pax7 突变卫星细胞的细胞和分子缺陷。何时以及如何做 Pax7突变细胞在发育到成体的过渡过程中出现缺陷？我将结合大量的检测 用于确定多种肌肉干细胞特性，包括肌肉分化、细胞增殖、细胞 死亡、生态位占据和细胞极性。这些测定将通过时间过程研究进行 新生儿到成人。 目标 2：对 Pax7 对照和 Pax7 突变卫星细胞进行 mRNA 测序。什么是转录组 在此过渡期间发生的变化以及 Pax7 管辖的具体流程是什么？比较时间 在整个发育过程中对照和 Pax7 突变细胞之间的转录组研究 将使用 mRNA-seq 进行过渡期，然后进行生物信息学分析。 目标 3：筛选候选基因在卫星细胞中的功能。基因是什么（除了 Pax7）对于发育成成人肌肉干细胞的转变至关重要？从目标 2 中，我将选择最多 50 个基因 测试它们在这方面的功能；它们可以独立行动，也可以在 Pax7 下游行动。我会发展 一种通过基因敲除和肌肉注射相结合来快速筛选这些基因功能的新方法 肌肉干细胞移植。