Endothelial Barrier Protection and Repair in Acute Lung Injury

描述(由申请者提供)：本项目的重点是血管内皮细胞(EC)通透性在急性肺损伤(ALI)中的作用。该计划项目赠款的总体目标是更好地了解EC RhoA/RAC 1平衡紊乱的潜在机制，以及开发在ALI发展过程中恢复屏障完整性的治疗剂。该方案项目由一个高生产率的专家团队领导，由四个相互关联的项目和三个核心组成。项目1将重点介绍蛋白质硝化作为一种新描述的机制在EC屏障破坏和ALI发展过程中调节RhoA和RAC1信号的作用。分子、细胞、生化和整体动物研究将进一步阐明硝化作用改变RhoA和Rac 1核苷酸循环的机制，同时开发新的试剂来恢复ALI期间RhoA/Rac 1的平衡。项目2与项目1主题相联系，重点是Hsp90在调节RhoA激活和下游RhoA信号中的作用，并将比较和对比广泛基础的Hsp90抑制剂与靶向特定蛋白质相互作用的诱骗多肽的治疗可能性。项目3将继续关注RhoA/Rac 1平衡，并将重点放在腺苷诱导的Rac 1激活的治疗效果上，并将阐明Rac 1的新下游效应，并确定在体外和体内调节其表达的治疗潜力。项目1-3将主要集中在G-毒素，内毒素和项目4将完成我们的研究，以调查G-I-孔形成毒素、溶气素和溶血素对屏障的破坏作用。项目4中的研究将阐明RhoA/RAC1失衡的机制，并重点关注增强的NO信号在G介导的ALI期间恢复这种平衡的治疗潜力。两个科学核心(人类细胞与动物和分析)和一个行政核心以具有成本效益的方式提供多项目支持、专业知识和服务，大大加强了整个科学计划。预计使用最先进的细胞、分子、生化和生理学方法的高度集成的项目不仅将增加我们对RhoA和Rac 1在革兰氏阴性和革兰氏阳性诱导的ALI期间被调控的机制的理解，而且将促进开发新的战略和目标来治疗这种40年来死亡率没有显著下降的疾病。

DESCRIPTION (provided by applicant): This Program project is focused on the role of vascular endothelial cell (EC) permeability as a key component in acute lung injury (ALI). The overall goals of this Program Project Grant are to develop a better understanding of the mechanisms underlying disturbances in EC RhoA/Rac 1 balance and in developing therapeutic agents that restore barrier integrity during the development of ALI. The Program Project is led by a team of highly productive experts and comprised of four inter-related projects and 3 cores. Project 1 will focus on the role of protein nitration as a newly described mechanism to regulate RhoA and Rac 1 signaling during the development of EC barrier disruption and ALI. Molecular, cellular, biochemical, and whole animal studies are proposed to further elucidate the mechanisms by which nitration alters RhoA and Rac 1 nucleotide cycling while developing novel reagents to restore the RhoA/Rac 1 balance during ALI. Project 2 is thematically linked to Project 1 by focusing on the role of Hsp90 in regulating RhoA activation and downstream RhoA signaling and will compare and contrast the therapeutic possibilities of broad-based Hsp90 inhibitors with decoy peptides that target specific protein interactions. Project 3 continues the focus on RhoA/Rac 1 balance and will focus on the therapeutic effects of adenosine-induced Rac 1 activation and will elucidate new downstream effectors of Rac 1 and determine the therapeutic potential of modulating their expression both in vitro and in vivo. Projects 1-3 will focus predominantly on the G- toxin, LPS and Project 4 will round out our studies to investigate the barrier disruptive effects of the G-i- pore forming toxins, pneumolysin and Lysteriolysin. Studies in Project 4 will elucidate mechanisms of RhoA/Rac 1 imbalance and focus on the therapeutic potential of enhanced NO signaling in restoring this balance during G-imediated ALI. Two scientific cores (Human Cell & Animal and Analytical) and an Administrative Core provide multi-project support, expertise and service in a cost-effective manner leading to significant strengthening of the entire scientific Program. It is anticipated that the highly integrated Projects using stateof- the-art cellular, molecular, biochemical, and physiological approaches that will not only increase our understanding of the mechanisms by which RhoA and Rac 1 are regulated during both gram negative- and gram positive-induced ALI but will facilitate the development of new strategies and targets for the treatment of a disease that has not seen a significant drop in mortality in 40 years.