Directing the Fate of Cells to Myogenic Lineages

将细胞的命运引导至肌源性谱系

• 批准号: 7029248
• 负责人: MINDY GEORGE-WEINSTEIN
• 金额: $ 27.71万
• 依托单位: PHILADELPHIA COLLEGE OF OSTEOPATHIC MED
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7029248
• 关键词: Wnt gene /protein; biological signal transduction; bone morphogenetic proteins; cadherins; cardiac myocytes; cell adhesion; cell cell interaction; cell communication molecule; cell differentiation; cell growth regulation; chick embryo; embryogenesis; gene induction /repression; growth inhibitors; immunofluorescence technique; mesoderm; myoblasts; myocardium; myogenesis; nonmammalian vertebrate embryology; pluripotent stem cells; polymerase chain reaction; striated muscles; tissue /cell culture; transcription factor; transforming growth factors; western blottings

DESCRIPTION (provided by applicant): Cell-based regenerative medicine is a natural extension of developmental biology. Embryonic stem cells (ES cells) give rise to the entire embryo and have the potential to repair diseased tissues in the adult. Cell differentiation is preceded by specification, a process that restricts these pluripotent cells to a particular lineage. Tissue regeneration is dependent on the availability of factors that specify stem cells to the desired lineage and promote their differentiation. Given that the environment of diseased tissues is unlikely to be the same as that of the developing embryo, stem cells may require "conditioning" prior to or during implantation. The overall hypotheses guiding the proposed research are: (1) the fate of pluripotent stem cells can be directed in vitro and in vivo by stem cells previously committed to a particular lineage, and (2) the induction process is mediated by a combination of soluble factors and cell-cell adhesions. This project builds on our published observations that chick epiblast cells expressing MyoD mRNA in vivo recruit pluripotent epiblast cells to the skeletal muscle lineage in vitro by releasing a factor(s) that inhibits the bone morphogenetic protein (BMP) signaling pathway and promotes the expression of N-cadherin. In the absence of MyoD positive epiblast cells, more pluripotent cells form cardiac muscle. A key event in muscle differentiation is the downregulation of E-cadherin. Preliminary studies demonstrate that ablation of MyoD positive cells in the epiblast results in herniation of the organs through the ventral body wall and a reduction in skeletal muscle in the somites. The proposed studies are designed to explore the mechanism whereby MyoD positive epiblast cells promote skeletal myogenesis in the somites in vivo, and to identify the factors that regulate cadherin switching and recruitment of cells to the skeletal and cardiac muscle lineages in vitro. The experiments will focus on the roles of Wnt, 7GF-/3, BMP, and BMP inhibitor family members in these processes. The proposed studies may reveal a novel mechanism for regulating skeletal myogenesis in vivo, and lead to the development of methods for conditioning stem cells to ensure that they are biased to differentiate along the desired pathway and express the appropriate cell-cell adhesion proteins prior to their implantation into diseased tissues.

描述（由申请人提供）：基于细胞的再生医学是发育生物学的自然扩展。胚胎干细胞（ES细胞）会产生整个胚胎，并有可能修复成人患病的组织。细胞分化之前是规格，该过程将这些多能细胞限制为特定谱系。组织再生取决于将干细胞指定为所需谱系并促进其分化的因素的可用性。鉴于患病组织的环境不太可能与发育中的胚胎的环境相同，因此干细胞可能在植入之前或期间需要“调节”。指导拟议的研究的总体假设是：（1）多能干细胞的命运可以通过先前致力于特定谱系的干细胞在体外和体内定向，并且（2）诱导过程是由可溶性因子和细胞细胞粘附的组合介导的。该项目以我们发表的观察结果为基础，即在体内表达Myod mRNA的小鸡培养多能细胞表达多能细胞通过释放骨形态遗传蛋白（BMP）信号通路并促进N-核辅助素表达的因子，从而在体外对骨骼肌谱系进行体外体外。在没有Myod阳性层细胞细胞的情况下，更多的多能细胞会形成心肌。肌肉分化的关键事件是电子钙粘蛋白的下调。初步研究表明，薄细胞中Myod阳性细胞的消融导致器官通过腹壁造成疝，并减少了体积的骨骼肌。拟议的研究旨在探索MYOD阳性层细胞细胞在体内促进骨骼肌发生的机制，并确定调节钙粘蛋白转换和募集细胞向骨骼和心肌谱系的因素。实验将重点关注Wnt，7GF-/3，BMP和BMP抑制剂家族成员在这些过程中的作用。拟议的研究可能揭示了调节体内骨骼肌发生的新机制，并导致开发用于调节干细胞的方法，以确保它们沿沿所需途径区分并表达适当的细胞 - 细胞粘附蛋白在将其植入患病组织之前具有偏差。