CARDIAC PROGENITOR CELL DIFFERENTIATION

心脏祖细胞分化

• 批准号: 5991501
• 负责人: MAQ A SIDDIQUI
• 金额: $ 27.09万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/5991501
• 关键词: binding sites; cell differentiation; developmental genetics; gene targeting; genetic promoter element; genetically modified animals; laboratory mouse; mammalian embryology; myocardium; protein protein interaction; stem cells; transcription factor

DESCRIPTION: (Adapted from the Investigator's Abstract) The applicant hopes to understand the role of cardiogenic transcription factors in the determination and development of cardiac muscle cells. He has identified and cloned a cDNA encoding a cardiac regulatory transcription factor, called Clp-1, that may play an important role in the development of cardiogenic progenitor cells. The stated goal of the current application is to understand the role of Clp-1 in cardiogenesis by addressing when, where and in what capacity Clp-1 performs its function as a regulator of cardiogenesis. The proposal has four specific aims. Aim 1 is directed towards additional characterization of the structural and functional properties of Clp-1 as it relates to its potential cardiogenic regulatory properties. The applicant and his colleagues will attempt to identify the structural domains of Clp-1 that are responsible for binding the B element MEF-2 binding site in the cardiac MLC-2 gene promoter and determine if these same structural domains mediate Clp-1 activation of the cardiac MLC-2 gene in vivo. Aim 2 is directed towards investigating the cellular functions of Clp-1 in order to establish its role as a regulator of cardiac genes and cardiogenic cell differentiation. These studies will focus on determining 1) the potential of Clp-1 to act as a cardiogenic transcriptional activator capable of participating in the activation of cardiac lineage-determining genes, 2) the ability of Clp-1 to irreversibly commit precursor cells to the cardiogenic cell lineage, and 3) the hierarchal relationship of Clp-1 to BMP-2, another developmental signal which is known to be important in commitment of stem cells to the cardiac lineage. Aim 3 will focus on the characterization of promoter elements required for control of Clp-1 expression. The applicant will examine the promoter region of the mouse Clp-1 gene in an effort to define the sequence(s) required for directing expression in cardiogenic mesodermal cells. Aim 4 will attempt to determine the importance and necessity of Clp-1 in the actual elaboration of the cardiogenic developmental program in vivo. This will be evaluated in knockout mice following genetic ablation of the Clp-1 locus. The development of these mice will be followed primarily with respect to the morphological and histological status of the developing heart and secondarily with regard to non-cardiac phenotypes should they arise. Abnormal heart development will be analyzed at the molecular level by assaying for the expression of normal heart genetic markers presumably under the control of the Clp-1 transcription factor. In addition, physiological analysis of transgenic adult hearts will be undertaken to look for subtle perturbations that lead to detectable changes in heart function.

描述：（改编自研究者摘要）申请人希望 了解心原性转录因子在测定中的作用 和心肌细胞的发育。他鉴定并克隆了一个 cDNA 编码称为 Clp-1 的心脏调节转录因子，可能发挥作用 在心原性祖细胞的发育中发挥重要作用。这 当前应用程序的既定目标是了解 Clp-1 在 通过解决 Clp-1 何时、何地以及以什么能力发挥作用来实现心脏发生 作为心脏发生的调节剂。该提案有四个具体目标。 目标 1 旨在进一步表征结构和 Clp-1 的功能特性与其潜在的心源性相关 监管属性。申请人和他的同事将尝试 识别负责结合 B 的 Clp-1 结构域 心脏 MLC-2 基因启动子中的元件 MEF-2 结合位点，并确定是否 这些相同的结构域介导心脏 MLC-2 的 Clp-1 激活 体内的基因。目标 2 旨在研究细胞功能 Clp-1 以确定其作为心脏基因调节剂的作用 心肌细胞分化。这些研究将重点确定 1) Clp-1 作为心源性转录激活剂的潜力 能够参与心脏谱系决定的激活 基因，2) Clp-1 不可逆地将前体细胞定向到 心源性细胞谱系，以及 3) Clp-1 与 BMP-2 的层级关系， 众所周知，另一个对承诺很重要的发展信号 干细胞进入心脏谱系。目标 3 将重点关注 控制 Clp-1 表达所需的启动子元件。申请人将 检查小鼠 Clp-1 基因的启动子区域，以确定 指导心源性中胚层细胞表达所需的序列。 目标 4 将尝试确定 Clp-1 在 体内心源性发育程序的实际阐述。这将 在 Clp-1 基因座基因消除后，在基因敲除小鼠中进行评估。 主要关注这些小鼠的发育情况 发育中心脏的形态和组织学状态，其次 关于非心脏表型是否出现。心脏异常 将通过分析在分子水平上分析发育 正常心脏遗传标记的表达可能是在 Clp-1 转录因子。此外，转基因的生理分析 成年人的心脏将被用来寻找导致 心脏功能可检测到的变化。