Therapeutic Mechanisms of RhoA Inhibition In Acute Lung Injury

抑制 RhoA 治疗急性肺损伤的机制

基本信息

批准号：
8198063
负责人：
John D Catravas
金额：
$ 35.02万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-10 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8198063
关键词：
17-(Allylamino)-17-demethoxygeldanamycin Acute Acute Lung Injury Adenosine Adenoviruses Adult Respiratory Distress Syndrome Adverse effects Bacteria Bacterial Toxins Binding Blood capillaries Capillary Permeability Client Collaborations Combined Modality Therapy Data Diagnosis Endothelial Cells Exhibits F-Actin Functional disorder Genes Goals Gram-Negative Bacteria HSP 90 inhibition Heat Shock Protein 27 Heat-Shock Proteins 90 Human In Vitro Incidence Instruction Lung Lung Inflammation Malignant Neoplasms Mediating Mus Outcome Peptides Permeability Phase II Clinical Trials Phosphorylation Phosphotransferases Proteins Publishing Reagent Regulation Research Personnel Research Subjects Respiratory physiology Role Scientist Severities Signal Transduction Stress Fibers Testing Therapeutic base capillary in vivo inhibitor/antagonist mortality mouse model nitration novel novel strategies prevent repaired research study

项目摘要

PROJECT SUMMARY (See instructions); Subjects diagnosed with ARDS exhibit ~40% mortality, thus demanding fresh approaches to the management of this serious condition. This Project will investigate a totally new mechanism of both G+ and G- bacteria-induced endothelial barrier dysfunction in human endothelial cells and in ALI/ARDS. It follows the studies on the crucial role RhoA/Rac1 imbalance in endothelial barrier dysfunction and ALI/ARSD, proposed in Project 1. Here we focus on the activation of endothelial RhoA that produces endothelial hyperpermeability in culture and increased pulmonary capillary permeability, in vivo. We propose a novel mechanism of regulating RhoA activity that could be useful in the management of ALI and ARDS. Preliminary data from Project 1 and published studies suggest that pp60src is a key step in RhoA activation, which leads to the phosphorylation of the small heat shock protein 27 (hsp27), a major cause of F-actin stress fiber formation and endothelial barrier dysfunction. The kinase, ppGOsrc is a well-known heat shock protein 90 (hsp90) client protein and we recently published that hsp27 co-immunoprecipitates with hsp90. Preliminary data suggest that the hsp90 inhibitor, 17-/\AG reduces both LPS-induced ppSOsrc activation and hsp27 phosphorylation, as well as LPS-induced RhoA activation, in endothelial cells. Furthermore, we have recently demonstrated that hsp90 inhibition prevents and reverses LPS-induced endothelial barrier dysfunction, in culture, and reduces capillary hyper-permeability, inflammation, lung dysfunction and mortality in a mouse model of LPS-induced ALI. Still, the effects of hsp90-mediated regulation of ppBOsrc, RhoA and hsp27 activation on G+ and G- induced endothelial barrier dysfunction, especially human endothelial cell barrier function, and in the management of ALI, in vivo, remain unknown. This project will test the hypothesis that hsp90 is an important regulator of human endothelial cell hyper-permeability, in vitro, and of ALI, in vivo. We will further test the hypothesis that hsp90 exerts these actions, in part, by controlling the fate of two key proteins (ppSOsrc, hsp27) that are involved in RhoA activation and signal transduction. These studies represent an exciting new possibility in the management of ALI/ARDS.

项目摘要（请参阅说明）；被诊断为ARDS的受试者表现出约40％的死亡率，因此要求对这种严重疾病的管理采取新的方法。该项目将研究人类内皮细胞和ALI/ARDS中G+和G-细菌诱导的内皮屏障功能障碍的全新机制。它遵循有关在项目1中提出的内皮屏障功能障碍和ALI/ARSD中关键作用RHOA/RAC1失衡的研究。在这里，我们专注于在培养中产生内皮过度过敏性的激活，并在Vivo中产生内皮过度过敏性，并增加了肺毛细管透化性。我们提出了一种调节RHOA活性的新型机制，该机制可用于ALI和ARDS的管理。项目1和已发表的研究的初步数据表明，PP60SRC是RhoA激活的关键步骤，它导致小热休克蛋白27（HSP27）的磷酸化，这是F-肌动蛋白应激纤维形成和内皮屏障功能障碍的主要原因。激酶PPGOSRC是一种众所周知的热激蛋白90（HSP90）客户蛋白，我们最近发表了HSP27与HSP90共免疫沉淀。初步数据表明，HSP90抑制剂17-/\ ag降低了LPS诱导的PPSOSRC激活和HSP27磷酸化以及在内皮细胞中LPS诱导的RHOA激活。此外，我们最近证明，HSP90抑制作用可防止并逆转LPS诱导的内皮屏障功能障碍，培养和降低毛细血管超渗透性，炎症，肺功能障碍和LPS诱导ALI小鼠模型中的死亡率。尽管如此，HSP90介导的PPBOSRC，RHOA和HSP27激活对G+和G-诱导的内皮屏障功能障碍的影响，尤其是人体内皮细胞屏障功能，以及在体内Ali的管理中，尚不清楚。该项目将检验以下假设：HSP90是体内人类内皮细胞过度过度性的重要调节剂和体内Ali的重要调节剂。我们将进一步检验以下假设：HSP90部分通过控制参与RHOA激活和信号转导的两个关键蛋白（PpsosRC，HSP27）的命运来部分发挥这些作用。这些研究代表了ALI/ARDS管理的一种令人兴奋的新可能性。