Endothelial Barrier Protection and Repair in Acute Lung Injury

描述（由申请人提供）：本项目重点研究血管内皮细胞（EC）通透性在急性肺损伤（ALI）中的关键作用。该项目的总体目标是更好地了解EC RhoA/Rac 1平衡紊乱的机制，并开发治疗药物，恢复ALI发展过程中的屏障完整性。该计划项目由一个高效率的专家团队领导，由四个相互关联的项目和3个核心组成。项目1将重点关注蛋白质硝化作为一种新描述的机制，在EC屏障破坏和ALI的发展过程中调节RhoA和Rac 1信号的作用。分子、细胞、生化和全动物研究将进一步阐明硝化改变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在主题上与项目1相关联，重点关注Hsp90在调节RhoA激活和下游RhoA信号传导中的作用，并将比较和对比广泛的Hsp90抑制剂与靶向特定蛋白质相互作用的诱饵肽的治疗可能性。项目3继续关注RhoA/Rac 1平衡，并将关注腺苷诱导的Rac 1激活的治疗效果，并将阐明Rac 1的新下游效应物，并确定在体外和体内调节其表达的治疗潜力。项目1-3将主要关注G-毒素，LPS和项目4将完善我们的研究，以研究G-i孔形成毒素，溶血素和溶菌素的屏障破坏作用。项目4的研究将阐明RhoA/Rac 1失衡的机制，并重点研究增强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，从而显著加强整个科学计划。预计高度整合的项目将使用最先进的细胞、分子、生化和生理学方法，这不仅将增加我们对在革兰氏阴性和革兰氏阳性诱导的ALI中RhoA和Rac 1调节机制的理解，而且将促进开发新的策略和靶点来治疗这种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.