Increased microvascular permeability causes pulmonary edema in acute lung injury, adult respiratory distress syndrome (ARDS), and multisystem organ failure. Disruption of endothelial cell-cell contacts is a possible cause of increased microvascular permeability. Identification of intracellular mechanisms which regulate changes in vascular permeability may lead to therapeutic strategies for controlling tissue damage due to sepsis or other causes of acute lung injury. Our preliminary results indicate that overexpression of protein kinase Cdelta (PKCDELTA) protects against thrombin-induced increases in endothelial monolayer permeability. The hypothesis of this proposal is that PKCDelta blunts changes in endothelial monolayer permeability by modulating the organization of cytoskeleton junctional complexes. Endothelial cells (EC) normally form a tightly adherent monolayer. During an inflammatory or thrombogenic response endothelial monolayer permeability increases with the formation of interendothelial cell gaps as a result of both EC contraction and disruption of endothelial cell-cell interactions. EC contraction occurs by intracellular calcium mobilization, Rho GTPase activation, myosin light chain phosphorylation, actin microfilament reorganization, and focal adhesion complex (FA) formation. Disruption of endothelial cell-cell junctions occurs concomitantly with the redistribution of the adherens junction (AJ) protein complexes at the points of intercellular gap formation. The intracellular mechanisms by which edemagenic agents promote EC contraction and FA formation and the disassembly of the AJ complexes are not completely understood. A number of inflammatory and thrombogenic agents activate PKC, increasing monolayer endothelial permeability. The overall objective of this proposal is to determine the mechanism by which PKCDELTA modulates endothelial barrier function. Using microvascular EC which stably overexpress PKCDELTA and vector control, we will determine: I) whether diminished endothelial barrier function is due to the stabilization and association of AJ or FA with PKCDELTA; II) whether delta PKCDELTA blunts endothelial barrier function by modulating EC retraction at the level of actin stress fiber formation and/or function; and III) whether PKCDELTA reduces endothelial barrier function by altering MAPK signaltransduction pathway. The elucidation of the molecular mechanisms by which PKCDELTA decreases endothelial monolayer permeability may lead to the development of therapeutic agents which are protective against endothelial barrier dysfunction.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL067795-01
Application #
6359745
Study Section
Lung Biology and Pathology Study Section (LBPA)
Program Officer
Garfinkel, Susan J
Project Start
2001-08-01
Project End
2005-07-31
Budget Start
2001-08-01
Budget End
2002-07-31
Support Year
1
Fiscal Year
2001
Total Cost
$252,000
Indirect Cost
Name
Brown University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
001785542
City
Providence
State
RI
Country
United States
Zip Code
02912
Harrington, Elizabeth O; Vang, Alexander; Braza, Julie et al. (2018) Activation of the sweet taste receptor, T1R3, by the artificial sweetener sucralose regulates the pulmonary endothelium. Am J Physiol Lung Cell Mol Physiol 314:L165-L176
Chichger, Havovi; Braza, Julie; Duong, Huetran et al. (2016) Select Rab GTPases Regulate the Pulmonary Endothelium via Endosomal Trafficking of Vascular Endothelial-Cadherin. Am J Respir Cell Mol Biol 54:769-81
Chichger, Havovi; Vang, Alexander; O'Connell, Kelly A et al. (2015) PKC ? and ?II regulate angiotensin II-mediated fibrosis through p38: a mechanism of RV fibrosis in pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol 308:L827-36
Chichger, Havovi; Braza, Julie; Duong, Huetran et al. (2015) Neovascularization in the pulmonary endothelium is regulated by the endosome: Rab4-mediated trafficking and p18-dependent signaling. Am J Physiol Lung Cell Mol Physiol 309:L700-9
Chichger, Havovi; Duong, Huetran; Braza, Julie et al. (2015) p18, a novel adaptor protein, regulates pulmonary endothelial barrier function via enhanced endocytic recycling of VE-cadherin. FASEB J 29:868-81
Chichger, Havovi; Braza, Julie; Duong, Huetran et al. (2015) SH2 domain-containing protein tyrosine phosphatase 2 and focal adhesion kinase protein interactions regulate pulmonary endothelium barrier function. Am J Respir Cell Mol Biol 52:695-707
Klinger, James R; Tsai, Shu-Whei; Green, Sabrina et al. (2013) Atrial natriuretic peptide attenuates agonist-induced pulmonary edema in mice with targeted disruption of the gene for natriuretic peptide receptor-A. J Appl Physiol (1985) 114:307-15
Grinnell, Katie L; Harrington, Elizabeth O (2012) Interplay between FAK, PKC?, and p190RhoGAP in the regulation of endothelial barrier function. Microvasc Res 83:12-21
Chichger, Havovi; Grinnell, Katie L; Casserly, Brian et al. (2012) Genetic disruption of protein kinase Cýý reduces endotoxin-induced lung injury. Am J Physiol Lung Cell Mol Physiol 303:L880-8
Grinnell, Katie L; Chichger, Havovi; Braza, Julie et al. (2012) Protection against LPS-induced pulmonary edema through the attenuation of protein tyrosine phosphatase-1B oxidation. Am J Respir Cell Mol Biol 46:623-32

Showing the most recent 10 out of 36 publications