title: Regulation of Thrombin-Induced Lung Endothelial Cell Barrier Disruption by Protein Kinase C Delta.
abstract: Protein Kinase C (PKC) plays a significant role in thrombin-induced loss of endothelial cell (EC) barrier integrity; however, the existence of more than 10 isozymes of PKC and tissue-specific isoform expression has limited our understanding of this important second messenger in vascular homeostasis. In this study, we show that PKCδ isoform promotes thrombin-induced loss of human pulmonary artery EC barrier integrity, findings substantiated by PKCδ inhibitory studies (rottlerin), dominant negative PKCδ construct and PKCδ silencing (siRNA). In addition, we identified PKCδ as a signaling mediator upstream of both thrombin-induced MLC phosphorylation and Rho GTPase activation affecting stress fiber formation, cell contraction and loss of EC barrier integrity. Our inhibitor-based studies indicate that thrombin-induced PKCδ activation exerts a positive feedback on Rho GTPase activation and contributes to Rac1 GTPase inhibition. Moreover, PKD (or PKCμ) and CPI-17, two known PKCδ targets, were found to be activated by PKCδ in EC and served as modulators of cytoskeleton rearrangement. These studies clarify the role of PKCδ in EC cytoskeleton regulation, and highlight PKCδ as a therapeutic target in inflammatory lung disorders, characterized by the loss of barrier integrity, such as acute lung injury and sepsis.
authors.affiliation: Institute for Personalized Respiratory Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Lishi
authors.initials: L
authors.lastname: Xie
authors.affiliation: Department of Medicine and Arizona Respiratory Center, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Eddie T
authors.initials: ET
authors.lastname: Chiang
authors.affiliation: Department of Medicine and Arizona Respiratory Center, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Xiaomin
authors.identifier: http://orcid.org/0000-0002-8792-0584
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Medicine and Arizona Respiratory Center, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Gabriel T
authors.initials: GT
authors.lastname: Kelly
authors.affiliation: Institute for Personalized Respiratory Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Prasad
authors.initials: P
authors.lastname: Kanteti
authors.affiliation: Department of Medicine, University of Chicago, Chicago, Illinois, United States of America.
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Singleton
authors.affiliation: Department of Medicine and Arizona Respiratory Center, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Sara M
authors.initials: SM
authors.lastname: Camp
authors.affiliation: Institute for Personalized Respiratory Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Tingting
authors.initials: T
authors.lastname: Zhou
authors.affiliation: Institute for Personalized Respiratory Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Dudek
authors.affiliation: Institute for Personalized Respiratory Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Pharmacology, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Viswanathan
authors.initials: V
authors.lastname: Natarajan
authors.affiliation: Department of Medicine and Arizona Respiratory Center, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: Department of Medicine and Arizona Respiratory Center, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Black
authors.affiliation: Institute for Personalized Respiratory Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine and Arizona Respiratory Center, University of Arizona, Tucson, Arizona, United States of America.
authors.forename: Joe G N
authors.initials: JG
authors.lastname: Garcia
authors.affiliation: Institute for Personalized Respiratory Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.|Department of Medicine, University of Illinois at Chicago, Chicago, Illinois, United States of America.
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Jacobson
mesh_terms: D000098:Acetophenones / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology; D042783:Endothelial Cells / Q000473:pathology*; D004789:Enzyme Activation / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008168:Lung / Q000201:enzymology* / Q000473:pathology*; D008954:Models, Biological; D009124:Muscle Proteins; D018994:Myosin Light Chains / Q000378:metabolism; D010749:Phosphoprotein Phosphatases; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology*; D020741:rho GTP-Binding Proteins / Q000378:metabolism
chemical_list: D000098:Acetophenones; D001578:Benzopyrans; D047908:Intracellular Signaling Peptides and Proteins; D009124:Muscle Proteins; D018994:Myosin Light Chains; C107507:PPP1R14A protein, human; C085746:rottlerin; C089212:protein kinase D; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D010749:Phosphoprotein Phosphatases; D013917:Thrombin; D020741:rho GTP-Binding Proteins