; 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.

Public Health Relevance

The goal of this study is to develop new approaches to the management of acute lung injury and acute respiratory distress syndrome, conditions that carry a high degree of mortality. We propose a novel approach aimed at restoring the known dysfunction and increased permeability of lung capillary vessels.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
1P01HL101902-01A1
Application #
8198063
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
2011-07-10
Project End
2016-06-30
Budget Start
2011-07-10
Budget End
2012-06-30
Support Year
1
Fiscal Year
2011
Total Cost
$350,150
Indirect Cost
Name
Georgia Regents University
Department
Type
DUNS #
966668691
City
Augusta
State
GA
Country
United States
Zip Code
30912
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Song, Shanshan; Ayon, Ramon J; Yamamura, Aya et al. (2017) Capsaicin-induced Ca(2+) signaling is enhanced via upregulated TRPV1 channels in pulmonary artery smooth muscle cells from patients with idiopathic PAH. Am J Physiol Lung Cell Mol Physiol 312:L309-L325
Wang, Ting; Gross, Christine; Desai, Ankit A et al. (2017) Endothelial cell signaling and ventilator-induced lung injury: molecular mechanisms, genomic analyses, and therapeutic targets. Am J Physiol Lung Cell Mol Physiol 312:L452-L476
Kumar, Sanjiv; Sun, Xutong; Noonepalle, Satish Kumar et al. (2017) Hyper-activation of pp60Src limits nitric oxide signaling by increasing asymmetric dimethylarginine levels during acute lung injury. Free Radic Biol Med 102:217-228
Chen, F; Wang, Y; Rafikov, R et al. (2017) RhoA S-nitrosylation as a regulatory mechanism influencing endothelial barrier function in response to G+-bacterial toxins. Biochem Pharmacol 127:34-45
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Rafikova, Olga; Rafikov, Ruslan; Kangath, Archana et al. (2016) Redox regulation of epidermal growth factor receptor signaling during the development of pulmonary hypertension. Free Radic Biol Med 95:96-111
Xie, Lishi; Chiang, Eddie T; Wu, Xiaomin et al. (2016) Regulation of Thrombin-Induced Lung Endothelial Cell Barrier Disruption by Protein Kinase C Delta. PLoS One 11:e0158865
Kovacs-Kasa, Anita; Gorshkov, Boris A; Kim, Kyung-Mi et al. (2016) The protective role of MLCP-mediated ERM dephosphorylation in endotoxin-induced lung injury in vitro and in vivo. Sci Rep 6:39018

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