Physiological regulation of vascular function by mutant caveolin-1

描述（由申请人提供）：内源性小窝蛋白-1 （Cav-1）对eNOS功能的抑制已经在体外和体内得到了很好的表征。我们进一步剖析了eNOS与Cav-1之间的相互作用，并通过诱变Cav-1支架结构域（CSD）解离了Cav-1与eNOS的抑制结合。AP-Cav 3PM是一种细胞渗透性肽，含有Cav-1 CSD的突变形式，在体外和离体激活eNOS。重要的是，我们现在已经开发出可诱导的内皮细胞特异性Cav-1转基因小鼠，这些小鼠具有Cav-1的突变形式（F92A Cav-1），通过Cav-1和eNOS的解离促进NO的释放。考虑到这一点，了解eNOS和Cav-1之间的相互作用将允许分子解剖Cav-1作为eNOS的负调节因子及其在血管生成中的作用。我们假设F92A Cav-1突变小鼠将表现出结构正常的小窝和Cav-1分布，此外还会增加NO的产生、降低血压、降低血管反应性和增加血管生成潜力。令人兴奋的是，这一提议提供了第一个体内平台来研究Cav-1作为小泡主要结构成分的生理作用与其作为信号传导平台的作用。为了更详细地研究这种重要相互作用的调节，我们将确定：1 . F92A Cav-1突变对体内心血管表型的影响；2. F92A Cav-1突变对体外和体外血管生理反应的影响F92A Cav-1突变如何影响体内血管生成。

DESCRIPTION (provided by applicant): Inhibition of eNOS function by endogenous caveolin-1 (Cav-1) has been well characterized in vitro and in vivo. We have further dissected the interaction between eNOS and Cav-1, and have dissociated the inhibitory binding of Cav-1 to eNOS using mutagenesis of the caveolin scaffolding domain (CSD). AP-Cav 3PM, a cell permeable peptide harboring a mutant form of the Cav-1 CSD, activates eNOS in vitro and ex vivo. Importantly we have now developed inducible, endothelial cell specific Cav-1 transgenic mice that have a mutant form of Cav-1 (F92A Cav-1) that promotes NO release through the dissociation of Cav-1 and eNOS. With this in mind, understanding the interactions between eNOS and Cav-1 will permit molecular dissection of the roles of Cav-1 as a negative regulator of eNOS and the role this plays angiogenesis. We hypothesize that F92A Cav-1 mutant mice will exhibit structurally normal caveolae and Cav-1 distribution, in addition to increased NO production, reduced blood pressure, lower vascular reactivity, and increased angiogenic potential. Excitingly, this proposal presents the first in vivo platform for the study of Cav-1 physiological role as the major structural component of caveolae versus its role as a signaling platform. To examine the regulation of this important interaction in more detail, we will determine:1 The effects of the F92A Cav-1 mutation on the cardiovascular phenotype in vivo; 2. The effects of the F92A Cav-1 mutation on physiologic responses of the vasculature ex vivo and 3. How the F92A Cav-1 mutation affects angiogenesis in vivo. PUBLIC HEALTH RELEVANCE: Project Narrative This research is relevant to public health since endothelial dysfunction is a common manifestation of most cardiovascular diseases. Our research will design new approaches to improve blood flow and reduce atherogenesis. Research supported by this grant may help identify pathways and molecular targets that reduce heart disease and improve the quality of life of people suffering with cardiovascular disease.