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功能的抑制作用已在体外和体内得到充分表征。我们进一步剖析了eNOS和Cav-1之间的相互作用，并使用小窝蛋白支架结构域（CSD）的诱变分离了Cav-1对eNOS的抑制性结合。AP-Cav 3 PM是一种具有Cav-1 CSD突变形式的细胞渗透性肽，在体外和离体激活eNOS。重要的是，我们现在已经开发了可诱导的内皮细胞特异性Cav-1转基因小鼠，其具有Cav-1的突变形式（F92 A Cav-1），其通过Cav-1和eNOS的解离促进NO释放。考虑到这一点，了解eNOS和Cav-1之间的相互作用将允许Cav-1作为eNOS的负调节因子的作用的分子解剖和这起血管生成的作用。我们假设F92 A Cav-1突变小鼠将表现出结构正常的小窝和Cav-1分布，除了增加NO的产生，降低血压，降低血管反应性，并增加血管生成的潜力。令人兴奋的是，该提案提出了第一个研究Cav-1生理作用的体内平台，作为小窝的主要结构成分，而不是作为信号平台。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 F92 A Cav-1突变对体内心血管表型的影响; 2. F92 A Cav-1突变对离体血管系统生理反应的影响和3. F92 A Cav-1突变如何影响体内血管生成 公共卫生相关性： 由于内皮功能障碍是大多数心血管疾病的常见表现，因此本研究与公共卫生相关。我们的研究将设计新的方法来改善血液流动和减少动脉粥样硬化。这项资助支持的研究可能有助于确定减少心脏病和改善心血管疾病患者生活质量的途径和分子靶点。

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.