The goal of this K08 application is to provide the PI with essential skills in which to be a successful academician and achieve independent scientific investigator status. The PI will focus upon the endothelium, a functionally dynamic cellular monolayer which performs critical roles in angiogenesis and as a semi-selective barrier between the vasculature and surrounding tissues. Despite its importance in acute lung injury syndromes, the cellular regulation of endothelial barrier function, especially barrier restoration and enhancement, is poorly understood. Recent work in endothelial cells (EC) demonstrates that phosphorylation of myosin light chains (MLC) by the enzyme myosin light chain kinase (MLCK) is critical to induction of vascular permeability in some models. Studies in our laboratory have revealed that EC express a novel, high molecular weight MLCK isoform that when tyrosine phosphorylated is found in stable complex with the actin-binding protein, cortactin, an actin-binding protein implicated in multiple aspects of cytoskeletal organization and rearrangement including Arp2/3-mediated actin polymerization. We hypothesize that cortactin plays an active and critical role in endothelial cell cytoskeletal rearrangement during barrier enhancement and migration. SA#1 will examine the role of cortactin in cultured pulmonary EC barrier function and migration using molecular biology techniques. SA#2 will explore the role of cortactin phosphorylation in pulmonary EC cytoskeletal regulation. SA#3 will characterize the role of cortactin-EC MLCK interaction in pulmonary endothelial cytoskeletal rearrangement through biochemical, immunohistochemical, and molecular biology techniques. Finally, SA#4 will investigate the effect of biophysical shear stress and cyclical stretch on pulmonary endothelial cortactin biology and regulation. Through these complementary techniques and coursework, the PI will develop new skills and generate novel data regarding EC barrier restoration/enhancement, and angiogenesis which may potentially lead to new therapies for the devastating consequences of pulmonary edema.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Clinical Investigator Award (CIA) (K08)
Project #
7K08HL070013-04
Application #
6914962
Study Section
Special Emphasis Panel (ZHL1-CSR-M (F2))
Program Officer
Colombini-Hatch, Sandra
Project Start
2002-08-19
Project End
2007-06-30
Budget Start
2005-09-01
Budget End
2006-06-30
Support Year
4
Fiscal Year
2005
Total Cost
$125,010
Indirect Cost
Name
University of Chicago
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
Arce, Fernando Teran; Whitlock, Jenny L; Birukova, Anna A et al. (2008) Regulation of the micromechanical properties of pulmonary endothelium by S1P and thrombin: role of cortactin. Biophys J 95:886-94
Dudek, S M; Camp, S M; Chiang, E T et al. (2007) Pulmonary endothelial cell barrier enhancement by FTY720 does not require the S1P1 receptor. Cell Signal 19:1754-64
Li, Yansong; Uruno, Takehito; Haudenschild, Christian et al. (2004) Interaction of cortactin and Arp2/3 complex is required for sphingosine-1-phosphate-induced endothelial cell remodeling. Exp Cell Res 298:107-21
Bogatcheva, Natalia V; Dudek, Steven M; Garcia, Joe G N et al. (2003) Mitogen-activated protein kinases in endothelial pathophysiology. J Investig Med 51:341-52
Dudek, Steven M; Birukov, Konstantin G; Zhan, Xi et al. (2002) Novel interaction of cortactin with endothelial cell myosin light chain kinase. Biochem Biophys Res Commun 298:511-9
O'Neill, C A; van der Vliet, A; Eiserich, J P et al. (1995) Oxidative damage by ozone and nitrogen dioxide: synergistic toxicity in vivo but no evidence of synergistic oxidative damage in an extracellular fluid. Biochem Soc Symp 61:139-52