Development of effective therapies for treatment of acute lung injury (ALI) and adult respiratory distress syndrome (ARDS) remains a challenging task. Many experimental models for testing of novel protective agents utilize preventive or concurrent treatment during ALI induction, while post-treatment represents more clinically relevant intervention. Such differences in the timing of drug administration may have dramatic impact on the efficiency of treatment and activation of specific molecular mechanisms directing resolution of ongoing injury in contrast to blocking onset of ALI by drug pretreatment. This proposal will fill this void and explore effects of post-treatment with FDA-approved prostacyclin (PC) analog iloprost in the in vitro and in vivo septic ALI models. Inflammation and increased endothelial cell (EC) permeability play a major role in the pathophysiology of ALI. During the previous cycle of this proposal, we characterized for the first time the molecular mechanisms of PC-mediated protection in aseptic model of ventilator induced lung injury. Our preliminary studies suggest potent protective effects of PC pretreatment against LPS-induced lung inflammation and vascular leak. This proposal will investigate effects of PC post-treatment in cell culture and animal models of septic ALI caused by Gram-positive heat-inactivated Staphylococcus Aureus bacteria (HKSA). We hypothesize that signaling by Rap1 GTPase plays a dual role in PC-induced acceleration of ALI resolution via promotion of EC barrier repair and suppression of inflammatory endothelial activation.
Aim -1 will evaluate effects of PC post-treatment and define a role of Rap1 in acceleration of barrier recovery in HKSA challenged EC.
Aim -2 will define molecular mechanisms downstream of Rap1 involved in EC barrier recovery. We will study a role of Rap1 effectors KRIT1 and RIAM in enhancement of EC adhesive structures and peripheral cytoskeleton essential for re-establishment of EC barrier.
Aim -3 will study a role of Rap1, KRIT1 and RIAM stimulation by PC post-treatment in downregulation of HKSA-induced pulmonary EC activation.
Aim -4 will elucidate specific role of Rap1, KRIT1 and Riam in PC-facilitated ALI recovery in vivo using """"""""loss of function"""""""" and """"""""gain of function"""""""" molecular approaches and mouse genetic models. These studies will characterize novel protective mechanisms and identify new protein targets for future therapies aimed at prevention of the pulmonary vascular barrier dysfunction associated with acute lung injury.

Public Health Relevance

Adult respiratory distress syndrome (ARDS) remains a major cause of morbidity and mortality with an overall mortality rate of 30-40%. The acute phase of lung injury is characterized by increased endothelial permeability and compromise of the blood-gas barrier, which allows an influx of protein-rich fluid into the air spaces, causing pulmonary edema. Development of effective therapies for ALI/ARDS treatment currently represents major a challenge. This study will investigate molecular mechanisms underlying protective effects of prostacyclin against pulmonary vascular endothelial dysfunction and characterize for the first time novel mechanisms of recovery in the model of Gram-positive septic ALI. These studies will expand our knowledge about molecular mechanisms leading to ALI resolution and may identify new targets for drug therapies.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL087823-05
Application #
8371434
Study Section
Respiratory Integrative Biology and Translational Research Study Section (RIBT)
Program Officer
Moore, Timothy M
Project Start
2007-04-01
Project End
2016-06-30
Budget Start
2012-08-15
Budget End
2013-06-30
Support Year
5
Fiscal Year
2012
Total Cost
$395,000
Indirect Cost
$145,000
Name
University of Chicago
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
Birukova, Anna A; Shah, Alok S; Tian, Yufeng et al. (2016) Dual role of vinculin in barrier-disruptive and barrier-enhancing endothelial cell responses. Cell Signal 28:541-51
Tian, Yufeng; Gawlak, Grzegorz; O'Donnell 3rd, James J et al. (2016) Activation of Vascular Endothelial Growth Factor (VEGF) Receptor 2 Mediates Endothelial Permeability Caused by Cyclic Stretch. J Biol Chem 291:10032-45
Gawlak, Grzegorz; Son, Sophia; Tian, Yufeng et al. (2016) Chronic high-magnitude cyclic stretch stimulates EC inflammatory response via VEGF receptor 2-dependent mechanism. Am J Physiol Lung Cell Mol Physiol 310:L1062-70
Wu, Congqing; Huang, Ru-Ting; Kuo, Cheng-Hsiang et al. (2015) Mechanosensitive PPAP2B Regulates Endothelial Responses to Atherorelevant Hemodynamic Forces. Circ Res 117:e41-53
Birukova, Anna A; Meng, Fanyong; Tian, Yufeng et al. (2015) Prostacyclin post-treatment improves LPS-induced acute lung injury and endothelial barrier recovery via Rap1. Biochim Biophys Acta 1852:778-91
Meng, Fanyong; Mambetsariev, Isa; Tian, Yufeng et al. (2015) Attenuation of lipopolysaccharide-induced lung vascular stiffening by lipoxin reduces lung inflammation. Am J Respir Cell Mol Biol 52:152-61
Meliton, Angelo Y; Meng, Fanyong; Tian, Yufeng et al. (2015) Oxidized phospholipids protect against lung injury and endothelial barrier dysfunction caused by heat-inactivated Staphylococcus aureus. Am J Physiol Lung Cell Mol Physiol 308:L550-62
Meliton, Angelo; Meng, Fanyong; Tian, Yufeng et al. (2015) Role of Krev Interaction Trapped-1 in Prostacyclin-Induced Protection against Lung Vascular Permeability Induced by Excessive Mechanical Forces and Thrombin Receptor Activating Peptide 6. Am J Respir Cell Mol Biol 53:834-43
Poroyko, Valeriy; Meng, Fanyong; Meliton, Angelo et al. (2015) Alterations of lung microbiota in a mouse model of LPS-induced lung injury. Am J Physiol Lung Cell Mol Physiol 309:L76-83
Mambetsariev, Isa; Tian, Yufeng; Wu, Tinghuai et al. (2014) Stiffness-activated GEF-H1 expression exacerbates LPS-induced lung inflammation. PLoS One 9:e92670

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