Regulators of Development and Quiescence in the Human Muscle Stem Cell Lineage

人类肌肉干细胞谱系发育和静止的调节因子

• 批准号: 10239080
• 负责人: OLIVIER POURQUIE
• 金额: $ 60.95万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-25 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10239080
• 关键词: Adult; Basal lamina; Biological; Biological Assay; Biology; Cell Differentiation process; Cell Lineage; Cell Therapy; Cell Transplantation; Cell physiology; Cells; Characteristics; Chick Embryo; Complex; Data; Decision Making; Development; Disease; Embryo; Exhibits; Genes; Goals; HDAC1 gene; Hematopoietic stem cells; Heterogeneity; Histone Deacetylase Inhibitor; Human; Human Development; Image; In Vitro; Knowledge; Laboratories; Light; Maintenance; Mediating; Methods; Minor; Molecular; Mus; Muscle; Muscle Fibers; Muscle satellite cell; Myoblasts; Myopathy; Natural regeneration; PAX7 gene; Pathway interactions; Pluripotent Stem Cells; Population; Population Heterogeneity; Process; Proliferating; Property; Protocols documentation; Regenerative capacity; Resolution; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Somatic Cell; Specific qualifier value; Striated Muscles; System; Testing; Tissues; Tretinoin; Work; adult stem cell; design; experimental study; fetal; fiber cell; imaging approach; imaging study; in vitro Model; in vivo; in vivo evaluation; induced pluripotent stem cell; insight; muscle regeneration; myogenesis; progenitor; regeneration potential; regenerative; satellite cell; self-renewal; single cell analysis; single cell sequencing; single-cell RNA sequencing; stem cells; transcriptome; virtual

SUMMARY/ABSTRACT Adult skeletal muscle is endowed with significant regenerative capacity due to the presence of adult stem cells, called satellite cells (SCs). Very little is known about the developmental origin of these cells in humans and most of our knowledge derives from work performed in mouse. SCs are rare and they cannot be amplified in unlimited numbers, as they lose their regenerative potential in vitro. These issues have constituted a major roadblock for the study of their regenerative properties in humans and for the development of translational applications, such as cell therapy. The discovery that somatic cell reprogramming can be used to generate virtually endless numbers of cells (iPSCs) has brought new hope for developing strategies allowing to study these processes in human cells. We have recently developed efficient protocols to differentiate mouse and human embryonic pluripotent stem cells (ES and iPS) into striated muscle fibers and SCs in vitro [1, 6]. The present proposal aims to take advantage of these in vitro models combined with in vivo studies in mice to understand the development of the SC lineage in humans. In mouse, SCs constitute a heterogeneous population comprising a minor fraction of dormant cells with important regenerative capacity and a faster dividing larger population. Whether this heterogeneity is conserved in human is not known. Moreover, the developmental origin of this heterogeneity is not understood. Thus an important aim of the proposed project is to understand the developmental basis of this heterogeneity. We will compare the heterogeneity of mouse Pax7+ myogenic precursors developing in vivo and in vitro using a newly introduced method of single cell sequencing (InDrop) which allows to sequence thousands of cells in one experiment. A parallel analysis will be performed comparing human SCs differentiated in vitro to fetal and adult human muscle tissue. These studies are expected to reveal the developmental trajectories and the heterogeneity of the Pax7+ subpopulations in mouse and human SCs. We will also take advantage of our in vitro myogenic differentiation systems to perform high-resolution live- imaging studies of the development of the PAX7+ lineage. We will characterize poorly documented aspects of SC differentiation such as their entry in quiescence and their contribution to myotubes. Finally, we have identified Retinoic Acid (RA) and its co-activator complex WHHERE as a pathway able to induce reversible quiescence in PAX7+ cells differentiated in vitro. The role of RA in the differentiation and the maintenance of the PAX7+ lineage has not been investigated and we propose to characterize its function in vitro and in vivo in developing and mature SCs. We expect our work to shed light on the development and function of the human SC lineage, which could have significant implications for the development of cell-based therapies for muscle diseases.

摘要/摘要 由于存在成年茎，成年骨骼肌具有显着的再生能力 细胞，称为卫星细胞（SCS）。这些细胞在人类中的发育起源知之甚少 我们的大多数知识都来自于在鼠标中执行的工作。 SC是罕见的，它们不能 由于它们在体外失去了再生潜力，因此被无限数量放大。这些问题有 构成了研究其在人类的再生特性的主要障碍 开发翻译应用，例如细胞疗法。 可以使用体细胞重编程来生成几乎无尽的单元格的发现 （IPSC）为制定策略带来了新的希望，可以研究人类细胞中的这些过程。 我们最近开发了有效的方案来区分小鼠和人类胚胎多能 在体外，干细胞（ES和IP）成分为横纹的肌肉纤维和SC [1，6]。目前的提议旨在采取 这些体外模型的优势与小鼠体内研究相结合，以了解 人类的SC血统。 在小鼠中，SC构成一个异质种群，其中包括一小部分休眠细胞 重要的再生能力和更快的人口分裂。这种异质性是 在人类中保守的是未知的。而且，这种异质性的发展起源不是 理解。因此，拟议项目的一个重要目的是了解这一点的发展基础 异质性。我们将比较小鼠pax7+肌原性前体的异质性 并使用新引入的单细胞测序方法（INDROP）进行体外，该方法允许对 一个实验中成千上万的细胞。将进行比较人类SC的平行分析 在体外与胎儿和成年人的肌肉组织分化。这些研究有望揭示 小鼠和人类SC中PAX7+亚群的发育轨迹和异质性。 我们还将利用我们的体外肌生成分化系统来实现高分辨率 PAX7+谱系发展的成像研究。我们将描述记录不足的方面 SC差异化，例如他们进入静止和对肌管的贡献。最后，我们有 鉴定出视黄酸（RA）及其共激活因子复合物作为能够诱导可逆的途径 PAX7+细胞中的静止在体外分化。 RA在差异化和维护中的作用 尚未研究PAX7+谱系，我们建议在体外和体内表征其功能 在开发和成熟的SC中。我们希望我们的工作能够阐明 人类SC谱系，这可能对基于细胞的疗法的发展具有重大影响 肌肉疾病。