P38alpha in Mouse Embryonic Stem Cells

小鼠胚胎干细胞中的 P38alpha

基本信息

批准号：
7268132
负责人：
YAN-LIN GUO
金额：
$ 21.26万
依托单位：
UNIVERSITY OF SOUTHERN MISSISSIPPI
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-07-31 至 2009-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7268132
关键词：
3-Dimensional Abnormal Endothelial Cell Affect Animal Model Biological Assay Blood Vessels Cardiovascular system Cell Adhesion Cell Differentiation process Cell Lineage Cell Proliferation Cell physiology Cells Collagen Defect Depth Development Disease ES Cell Line Embryo Embryonic Development Endothelial Cells Family Genes Goals Growth and Development function In Vitro Investigation Knock-out Knockout Mice Knowledge Link MAPK14 gene Mammalian Cell Mitogen-Activated Protein Kinases Mitogens Modeling Molecular Morphogenesis Morphology Mus Nature Physiological Processes Physiology Property Protein Isoforms Protein Kinase Proteins Range Regulation Role Series Somatic Cell Stimulus Structure System Testing Therapeutic Tube Vascular System Vascularization angiogenesis base cell type embryonic stem cell human MAPK14 protein in vivo leukemia inhibitory factor novel prevent stem tool

项目摘要

DESCRIPTION (provided by applicant): p38 mitogen activated protein (MAP) kinases are widely expressed protein kinases that regulate growth and development. p38alpha is the most abundant isoform expressed in mammalian cells. Depending on cell types and the nature of stimuli, p38alpha regulates a wide range of physiological processes, such as cell proliferation, survival, and differentiation. Its pivotal role in embryogenesis is best demonstrated in p38alpha knockout (p38a-/-) mice where embryos display defective vascularization and vessel structures, suggesting a critical role of p38alpha in the development of vascular system. p38alpha knockout is lethal, thus limited information can be obtained from animal models. However, the embryonic stem (ES) cells isolated from knockout embryos (p38a-/-ES cells) are viable, which can be a valuable cell system to analyze the specific functions of p38alpha. p38alpha has been intensively studied in somatic cells, but we know little of its functions in ES cells. We have shown that p38a-/-ES cells display several altered properties from wild type ES cells, including cell adhesion, morphology and viability. The goals of this proposal are: 1. to further characterize several lines of p38a and p38a-/-ES cells and to determine the molecular mechanisms underlying the altered properties of ES cells caused by p38alpha deletion, 2. to investigate the role of p38alpha in ES differentiation, specifically focusing on how the ability of ES cells to differentiate to endothelial cells (ECs) is affected in p38a-/-ES cell, and 3. to analyze ES cell-differentiated EC function by an 3-dimendsional (3D) in vitro angiogenesis model. In this model, ECs will be cultured in a 3D collagen matrix, where they undergo a series of morphological changes to form tube-like structures, mimicking the steps of in vivo blood vessel formation. This novel assay will be used as a functional analysis to test hypothesis that ECs derived from p38a-/-ES cells may retain certain abilities to differentiate, but differentiated cells may have impaired functions to assemble into normal vessels. This study is expected to provide well characterized and genetically defined pSSalpha deficiency ES cell lines, which will be particularly useful for in-depth investigation of the roles of pSSalpha in ES cell differentiation and vascular development. The knowledge derived from this study could be valuable for the development of cell-based therapeutic approaches for diseases associated with angiogenesis and cardiovascular malfunction.

描述（由申请人提供）：p38有丝分裂原活化蛋白（MAP）激酶是调节生长和发育的广泛表达的蛋白激酶。 p38alpha是在哺乳动物细胞中表达的最丰富的同工型。根据细胞类型和刺激的性质，p38Alpha调节了广泛的生理过程，例如细胞增殖，生存和分化。它在胚胎发生中的关键作用在p38Alpha敲除（p38a - / - ）小鼠中最好证明，其中胚胎表现出缺陷的血管化和血管结构，这表明p38alpha在血管系统发展中的关键作用。 p38Alpha敲除致命，因此可以从动物模型中获得有限的信息。但是，从敲除胚胎（p38a - / - ES细胞）分离的胚胎茎（ES）细胞是可行的，这可能是分析p38alpha的特定功能的有价值的细胞系统。 P38Alpha已在体细胞中进行了深入研究，但我们在ES细胞中的功能知之甚少。我们已经表明，p38a - / - ES细胞显示出来自野生型ES细胞的几种变化特性，包括细胞粘附，形态和生存能力。该提案的目标是：1。进一步表征p38a和p38a - / - ES细胞的几行，并确定由p38alpha删除引起的ES细胞改变性质的分子机制，2。 ES分化，特别关注ES细胞在p38a - / - ES细胞中如何影响ES细胞分化为内皮细胞（EC）的能力，以及3。分析ES细胞细胞分化的EC通过3维（3D）中的EC函数。体外血管生成模型。在该模型中，EC将在3D胶原蛋白基质中进行培养，在该基质中，它们经历了一系列形态的变化以形成类似管的结构，从而模仿体内血管形成的步骤。该新颖的测定法将用作功能分析，以检验源自p38a - / - ES细胞的EC可以保留某些能力以分化的能力，但是分化的细胞可能会受损的功能可以组装成正常血管。预计这项研究将提供良好的特征和遗传定义的PSSALPHA缺陷ES细胞系，这对于对PSSalpha在ES细胞分化和血管发育中的作用的深入研究特别有用。从这项研究中得出的知识对于开发与血管生成和心血管故障相关的疾病的基于细胞的治疗方法可能是有价值的。