Endothelial Cell Cytoskeletal Regulation by Cortactin

描述（由申请人提供）： 此K 08应用程序的目标是为PI提供以下方面的基本技能 这是一个成功的院士，实现独立的科学 研究者状态。PI将重点关注内皮，这是一种功能性的 动态细胞单层，在血管生成中起关键作用， 作为脉管系统和周围组织之间的半选择性屏障。 尽管其在急性肺损伤综合征中的重要性， 内皮屏障功能，特别是屏障恢复， 增强，了解甚少。内皮细胞（EC）的最新研究 证明了肌球蛋白轻链（MLC）的磷酸化的酶 肌球蛋白轻链激酶（MLCK）是诱导血管生成的关键 在某些模型中的渗透性。我们实验室的研究表明，EC 表达一种新的高分子量MLCK同种型，当酪氨酸 发现磷酸化的蛋白与肌动蛋白结合蛋白形成稳定的复合物， coronin，一种肌动蛋白结合蛋白，涉及多个方面， 包括Arp 2/3介导的肌动蛋白在内的细胞骨架组织和重排 聚合法我们假设corneumn在大脑中起着积极和关键的作用， 屏障增强过程中内皮细胞骨架重排的作用 和移民。SA#1将检查corneum在培养的肺细胞中的作用。 利用分子生物学技术研究EC屏障功能和迁移。SA#2 将探讨coronin磷酸化在肺EC中的作用 细胞骨架调节SA#3将描述皮质素-EC MLCK的作用 肺内皮细胞骨架重排相互作用 生物化学、免疫组织化学和分子生物学技术。最后， SA#4将研究生物物理剪切应力和周期性 牵张对肺内皮细胞生物学及调节作用。通过 这些补充技术和课程，PI将开发新的技能 并产生关于EC屏障恢复/增强的新数据，以及 血管生成，这可能会导致新的疗法，为破坏性的 肺水肿的后果。

DESCRIPTION (provided by applicant): The goal of this K08 application is to provide the PI with essential skills in which to be a successful academician and achieve independent scientific investigator status. The PI will focus upon the endothelium, a functionally dynamic cellular monolayer which performs critical roles in angiogenesis and as a semi-selective barrier between the vasculature and surrounding tissues. Despite its importance in acute lung injury syndromes, the cellular regulation of endothelial barrier function, especially barrier restoration and enhancement, is poorly understood. Recent work in endothelial cells (EC) demonstrates that phosphorylation of myosin light chains (MLC) by the enzyme myosin light chain kinase (MLCK) is critical to induction of vascular permeability in some models. Studies in our laboratory have revealed that EC express a novel, high molecular weight MLCK isoform that when tyrosine phosphorylated is found in stable complex with the actin-binding protein, cortactin, an actin-binding protein implicated in multiple aspects of cytoskeletal organization and rearrangement including Arp2/3-mediated actin polymerization. We hypothesize that cortactin plays an active and critical role in endothelial cell cytoskeletal rearrangement during barrier enhancement and migration. SA#1 will examine the role of cortactin in cultured pulmonary EC barrier function and migration using molecular biology techniques. SA#2 will explore the role of cortactin phosphorylation in pulmonary EC cytoskeletal regulation. SA#3 will characterize the role of cortactin-EC MLCK interaction in pulmonary endothelial cytoskeletal rearrangement through biochemical, immunohistochemical, and molecular biology techniques. Finally, SA#4 will investigate the effect of biophysical shear stress and cyclical stretch on pulmonary endothelial cortactin biology and regulation. Through these complementary techniques and coursework, the PI will develop new skills and generate novel data regarding EC barrier restoration/enhancement, and angiogenesis which may potentially lead to new therapies for the devastating consequences of pulmonary edema.