"Reversibility of Differentiating Myogenic Cells to Muscle Stem Cells"

“肌原细胞分化为肌肉干细胞的可逆性”

• 批准号: 8628048
• 负责人: CHEN-MING FAN
• 金额: $ 18.73万
• 依托单位: CARNEGIE INSTITUTION OF WASHINGTON, D.C.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8628048
• 关键词: Acute; Adult; Age; Aging-Related Process; Alleles; Animal Housing; Animal Model; Biological; Cells; Characteristics; Chimeric Proteins; Chronic; DNA; Data; Development; Direct Lytic Factors; Disease; Embryo; Estrogen Receptors; Event; Exercise; Funding; Gene Silencing; Genes; Genetic; Homeostasis; Injury; Investigation; Knock-in Mouse; Label; Mechanics; Methodology; Minor; Modeling; Mouse Strains; Movement; Mus; Muscle; Muscle Development; Muscle satellite cell; Myogenin; Myosin Light Chains; Natural regeneration; Nature; PTPRC gene; Pericytes; Perinatal; Posture; Process; Regulation; Reporter; Research; Research Personnel; Series; Skeletal Muscle; Source; Specificity; Staging; Tamoxifen; Testing; Time; Wasting Syndrome; Work; cell behavior; cell type; gene function; in vivo; injured; insight; muscle aging; muscle regeneration; muscular system; myogenesis; neonate; overexpression; progenitor; public health relevance; recombinase; stem cell population; tibialis anterior muscle; tool

DESCRIPTION (provided by applicant): The mouse provides an invaluable mammalian model for skeletal muscle research, partly due to continuous development of sophisticated genetic tools. The tamoxifen (tmx) inducible forms of Cre DNA recombinase (Cre) - estrogen receptor (ER) fusion protein is a powerful tool for inducible gene manipulation. A complete set of myogenic Cre-ERT2 (CE) alleles should facilitate the progress of the field. We have initiated making a series of myogenic CE KI alleles at the Pax3, Myf5, MyoD, Mrf4, Myogenin (Mgn), and Myosin light chain 1f (Mlc1f) loci (Aim 1). Together with Pax7, their expression represents a sequence of events, from muscle progenitor to terminally differentiated state, during development and regeneration. These new alleles will not only be useful for our research, but also beneficial to researchers in the field at large. There have been data implicating cell sources that do not express Pax7 (Pax7- cells) but act as muscle stem cells (e.g. CD45+Sca1+ cells, PICs, or pericytes). We hypothesize that these proclaimed Pax7- muscle stem cells are in fact myogenic cells that loses Pax7 expression in transit to differentiation but can regain Pax7 expression and return to the muscle stem cell state (Aim 2). Aim 1: Generating and characterizing a myogenic series of CE alleles. This series is under various stages of development. They will be used to perform tmx-inducible cell marking. Short-term cell marking will be performed to characterize the specificity of cell marking and the cell potential to contribute to muscles and other cell types. Aim 2: Testing the possibility that differentiating myogenic cells can revert to the muscle stem cell state. Two scenarios will be tested: 1) developmental progression and 2) regeneration. We will determine whether certain CE lines that do not label Pax7+ cells in short-term labeling, but give rise to Pax7+ cells after long term tracing, in either experimental paradigm. While the "reverted" stem cell population hypothesized may only represent a minor fraction, they likely have the potential to replenish the Pax7+ cells over time in chronic muscle wasting diseases and during the aging process.

描述（由申请人提供）：小鼠为骨骼肌研究提供了宝贵的哺乳动物模型，部分原因是复杂的遗传工具的不断发展。 Cre DNA 重组酶 (Cre) - 雌激素受体 (ER) 融合蛋白的他莫昔芬 (tmx) 诱导形式是诱导基因操作的强大工具。一套完整的肌源性 Cre-ERT2 (CE) 等位基因应该会促进该领域的进展。我们已开始在 Pax3、Myf5、MyoD、Mrf4、肌细胞生成素 (Mgn) 和肌球蛋白轻链 1f (Mlc1f) 位点（目标 1）制作一系列肌源性 CE KI 等位基因。它们与 Pax7 一起表达代表发育和再生过程中从肌肉祖细胞到终末分化状态的一系列事件。这些新的等位基因不仅对我们的研究有用，而且对整个领域的研究人员也有益。 有数据表明细胞来源不表达 Pax7（Pax7- 细胞），但充当肌肉干细胞（例如 CD45+Sca1+ 细胞、PIC 或周细胞）。我们假设这些宣称的 Pax7- 肌肉干细胞实际上是生肌细胞，在分化过程中失去 Pax7 表达，但可以重新获得 Pax7 表达并返回到肌肉干细胞状态（目标 2）。目标 1：生成并表征一系列肌源性 CE 等位基因。该系列正处于不同的开发阶段。它们将用于执行 tmx 诱导细胞标记。将进行短期细胞标记，以表征细胞标记的特异性以及细胞对肌肉和其他细胞类型做出贡献的潜力。目标 2：测试分化的生肌细胞恢复到肌肉干细胞状态的可能性。将测试两种情况：1）发育进程和2）再生。我们将确定某些 CE 系是否在短期标记中不标记 Pax7+ 细​​胞，但在任一实验范式中在长期追踪后产生 Pax7+ 细​​胞。虽然假设的“恢复”干细胞群可能只占一小部分，但随着时间的推移，它们可能有潜力在慢性肌肉萎缩疾病和衰老过程中补充 Pax7+ 细​​胞。

项目成果

A series of Cre-ER(T2) drivers for manipulation of the skeletal muscle lineage.

一系列用于操纵骨骼肌谱系的 Cre-ER(T2) 驱动程序。

• DOI: 10.1002/dvg.22792
• 发表时间: 2014
• 期刊: Genesis (New York, N.Y. : 2000)
• 作者: Southard,Sheryl;Low,SiewHui;Li,Lydia;Rozo,Michelle;Harvey,Tyler;Fan,Chen-Ming;Lepper,Christoph
• 通讯作者: Lepper,Christoph