Focal adhesion kinase in the cardiovascular system

心血管系统中的粘着斑激酶

• 批准号: 7028917
• 负责人: JUN-LIN GUAN
• 金额: $ 33.61万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-08 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7028917
• 关键词: angiogenesis; biological signal transduction; biopsy; cardiac myocytes; cardiovascular system; cell migration; cell proliferation; confocal scanning microscopy; drug adverse effect; embryo /fetus; focal adhesion kinase; gene targeting; genetically modified animals; growth factor receptors; heart failure; histogenesis; immunocytochemistry; integrins; laboratory mouse; mechanical stress; polymerase chain reaction; protein protein interaction; protein structure function; recombinase; vascular endothelium; ventricular hypertrophy

DESCRIPTION (provided by applicant): The long-term goals of this proposal are to understand the mechanisms and roles of focal adhesion kinase (FAK) and its signaling pathways in the cardiovascular system. FAK is a cytoplasmic tyrosine kinase that plays a key role in integrin-mediated signal transduction pathways as well as in signal transduction by growth factor receptors including those that play critical regulatory roles in the cardiovascular system. Consistent with these results from in vitro studies FAK gene knockout in mice resulted in an embryonic lethal phenotype with major defects in the axial mesodermal tissues and cardiovascular system. Despite the abundant knowledge of FAK interaction with other proteins and its roles in cell signaling in vitro, still relatively little is known about the in vivo Functions of FAK in embryonic development or in the adult organisms. Although the FAK total knockout mice suggested a potential role for FAK in the cardiovascular system, the embryonic lethality made it not very useful for studies on the role of FAK in cardiovascular development or functions in the adult animal. To overcome the problem of embryonic lethality of the FAK total knockout mice, we have generated FAK floxed mice with the FAK gene flanked by two loxP sites in preliminary studies. We have also generated transgenic mice with FAK transgene expression in the endothelial cells (ECs) and obtained transgenic mice with specific expression of Cre recombinase in ECs or cardiomyocytes. In this application, Aim 1 will investigate the role of FAK in ECs in [vasculogenesis and angiogenesis by breeding the FAK/IoxP mice with the TIE2-Cre mice to create EC-specific FAK knockout mice and examine the effects on vasculogenesis and angiogenesis in the embryo and in adult mice. Aim 2 will study molecular and cellular mechanism of FAK function in ECs by crossing the FAK conditional knockout mice with transgenic mice expressing FAK or its mutants and by isolating ECs from these mice to examine FAK interaction with other proteins, intracellular signaling pathways and cellular functions including migration and proliferation. Aim 3 will determine the role and mechanism of FAK in cardiac development and function by crossing FAK/loxP mice with cardiomyocyte specific Cre mice to generate heart specific FAK conditional knockout mice and analyzing the effects on the development and function of the heart. These studies will generate not only significant insights into the physiological function of FAK signaling pathways in vivo but also critical information for potential new therapies for pathological alterations in tumor angiogenesis and cardiac hypertrophy and heart failure.

描述(由申请人提供)： 这项建议的长期目标是了解粘着斑激酶(FAK)及其在心血管系统中的信号通路的机制和作用。FAK是一种胞质酪氨酸激酶，在整合素介导的信号转导通路以及生长因子受体的信号转导中发挥关键作用，其中生长因子受体在心血管系统中起着关键的调节作用。与体外研究的结果一致，小鼠的FAK基因敲除导致了胚胎致死表型，中轴中胚层组织和心血管系统存在主要缺陷。尽管对FAK与其他蛋白质的相互作用及其在体外细胞信号转导中的作用已经有了丰富的了解，但对于FAK在胚胎发育或成体生物体中的体内功能还知之甚少。尽管FAK基因完全敲除的小鼠暗示了FAK在心血管系统中的潜在作用，但胚胎的致死性使其对研究FAK在成年动物心血管发育或功能中的作用不是很有用。为了克服FAK全基因敲除小鼠胚胎致死的问题，我们在初步研究中产生了FAK基因两侧有两个loxP位点的FAK小鼠。我们还获得了在内皮细胞(ECs)中表达FAK的转基因小鼠，并获得了在ECs或心肌细胞中特异性表达Cre重组酶的转基因小鼠。在这项应用中，Aim 1将通过将FAK/IoxP小鼠与TIE2-CRE小鼠杂交，创建EC特异性FAK基因敲除小鼠，并检测其对胚胎和成年小鼠血管生成和血管生成的影响，来研究FAK在ECs中[血管生成和血管生成]中的作用。目的2通过将FAK条件基因敲除小鼠与表达FAK或其突变体的转基因小鼠杂交，并从这些小鼠分离内皮细胞，研究FAK与其他蛋白的相互作用、细胞内信号通路以及细胞功能(包括迁移和增殖)，以研究FAK在ECs中作用的分子和细胞机制。目的3通过将FAK/loxP小鼠与心肌细胞特异性Cre小鼠杂交，建立心脏特异的FAK条件性基因敲除小鼠，并分析其对心脏发育和功能的影响，以确定FAK在心脏发育和功能中的作用和机制。这些研究不仅将对FAK信号通路在体内的生理功能产生重要的见解，而且将为治疗肿瘤血管生成、心脏肥大和心力衰竭的潜在新疗法提供关键信息。