Protease activated receptor 1 (PAR-1) activation by thrombin is protective in human pulmonary artery endothelial cells if endothelial protein C receptor is occupied by its natural ligand

Jong-Sup Bae; Alireza R. Rezaie

doi:10.1160/TH08-02-0127

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2008; 100(01): 101-109
DOI: 10.1160/TH08-02-0127

Endothelium and Vascular Development

Schattauer GmbH

Protease activated receptor 1 (PAR-1) activation by thrombin is protective in human pulmonary artery endothelial cells if endothelial protein C receptor is occupied by its natural ligand

Jong-Sup Bae

¹Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, Missouri, USA

,

Alireza R. Rezaie

¹Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, Missouri, USA

› Institutsangaben

Abstract

Summary

We recently demonstrated that the occupancy of endothelial protein C receptor (EPCR) by its natural ligand activated protein C (APC)/protein C switches the protease activated receptor 1 (PAR-1)-dependent signaling specificity of thrombin from a disruptive to a protective effect in cultured human umbilical vein endothelial cells. Given the phenotypic differences between endothelial cells in venular and arterial beds, in this study we evaluated the signaling function of thrombin in human pulmonary artery endothelial cells (HPAECs) before and after treating them with PC-S195A which lacks catalytic activity but exhibits a normal affinity for EPCR. As expected, both thrombin and thrombin receptor agonist peptide (TRAP) enhanced the permeability barrier of HPA ECs; however, both PAR-1 agonists exhibited a potent barrier protective effect when the cells were treated with PC-S195A prior to stimulation by the agonists. Interestingly, similar toAPC, thrombin exhibited a potent cytoprotective activity in the LPS-induced permeability and TNF-α-induced apoptosis and adhesion assays in the PC-S195A treated HPAECs. Treatment of HPAECs with the cholesterol depleting molecule methyl-β-cyclodextrin eliminated the protective effect of both APC and thrombin. These results suggest that the occupancy of EPCR by its natural ligand recruits PA R-1 to a protective signaling pathway within lipid rafts of HPAECs. Based on these results we conclude that the activation of PAR-1 by thrombin would initiate a protective response in intact arterial vascular cells expressing EPCR. These findings may have important ramifications for understanding the mechanism of the participation of the vascular PAR-1 in pathophysiology of the inflammatory disorders.

Keywords

APC - thrombin - EPCR - PA R-1 - inflammation - signaling

Volltext

Referenzen

References
1 Esmon CT. Molecular events that control the protein C anticoagulant pathway. Thromb Haemost 1993; 70: 1-5.
2 Stenflo J. Structure and function of protein C. Sem Thromb Hemost 1984; 10: 109-121.
3 Walker FJ, Fay PJ. Regulation of blood coagulation by the protein C system. FASEB J 1992; 06: 2561-2567.
4 Dahlbäck B. Protein S and C4b-binding protein: Components involved in the regulation of the protein C anticoagulant system. Thromb Haemost 1991; 66: 49-61.
5 Esmon CT, Taylor Jr FB, Snow TR. Inflammation and coagulation: Linked processes potentially regulated through a common pathway mediated by protein C. Thromb Haemost 1991; 66: 160-165.
6 Joyce DE, Gelbert L, Ciaccia A. et al. Gene expression profile of antithrombotic protein C defines new mechanisms modulating inflammation and apoptosis. J Biol Chem 2001; 276: 11199-11203.
7 Mosnier LO, Griffin JH. The cytoprotective protein C pathway. Blood 2007; 109: 3161-3172.
8 Cheng T, Liu D, Griffin JH. et al. Activated protein C blocks p53-mediated apoptosis in ischemic human brain endothelium and is neuroprotective. Nature Medicine 2003; 09: 338-342.
9 Brueckman M, Horn S, Lang S. et al. Recombinant human activated protein C upregulates cyclooxygenase-2 expression in endothelial cells via binding to endothelial cell protein C receptor and activation of protease-activated receptor-1. Thromb Haemost 2005; 93: 743-750.
10 Nold MF, Nold-Petry CA, Fischer D. et al. Activated protein C downregulates p38 mitogen-activated protein kinase and improves clinical parameters in an in-vivo model of septic shock. Thromb Haemost 2007; 98: 1118-1126.
11 Bernard GR, Vincent JL, Laterre PF. et al. Efficacy and safety of recombinant human activated protein C for severe sepsis. N Eng J Med 2001; 344: 699-709.
12 Ruf W, Dorfleutner A, Riewald M. Specificity of coagulation factor signaling. J Thromb Haemost 2003; 01: 1495-1503.
13 Riewald M, Petrovan RJ, Donner A. et al. Activation of endothelial cell protease activated receptor 1 by the protein C pathway. Science 2002; 296: 1880-1882.
14 Ludeman MJ, Kataoka H, Srinivasan Y. et al. PAR1 cleavage and signaling in response to activated protein C andthrombin. J Biol Chem 2005; 280: 13122-13128.
15 Esmon CT. Is APC activation of endothelial cell PA R1 important in severe sepsis?: No. J Thromb Haemost 2005; 03: 1910-1911.
16 Bae J-S, Yang L, Rezaie AR. Receptors of the protein C activation and activated protein C signaling pathways are colocalized in lipid rafts of endothelial cells. Proc Natl Acad Sci USA 2007; 104: 2867-2872.
17 Bae J-S, Yang L, Manithody C. et al. The ligand occupancy of endothelial protein C receptor switches the PA R-1-dependent signaling specificity of thrombin from a permeability-enhancing to a barrier-protective response in endothelial cells. Blood 2007; 110: 3909-3916.
18 Garcia JGN, Pavalko FM, Patterson CE. Vascular endothelial cell activation and permeability responses to thrombin. Blood Coag Fibrinol 1995; 06: 609-626.
19 Finigan JH, Dudek SM, Singleton PA. et al. Activated protein C mediates novel lung endothelial barrier enhancement. J Biol Chem 2005; 280: 17286-17293.
20 Bode W, Mayr I, Baumann U. et al. The refined 1.9 Å crystal structure of human α-thrombin: interaction with D-Phe-Pro-Arg chlorometheylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO J 1989; 08: 3467-3475.
21 Kim I, Moon SO, Kim SH. et al. Vascular endothelial growth factor expression of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin through nuclear factor-kappa B activation in endothelial cells. J Biol Chem 2001; 276: 7614-7620.
22 Liu Y, Casey L, Pike LJ. Compartmentalization of phosphatidylinositol 4,5-bisphosphate in low-density membrane domains in the absence of caveolin. Biochem Biophys Res Commun 1998; 245: 684-690.
23 Feistritzer C, Riewald M. Endothelial barrier protection by activated protein C through PA R1-dependent sphingosine 1-phosphate receptor-1 crossactivation. Blood 2005; 105: 3178-3184.
24 Preston RJS, Ajzner E, Razzari C. et al. Multifunctional specificity of the protein C/activated protein C Gla domain. J Biol Chem 2006; 281: 28850-28857.
25 Ghosh S, Pendurthi UR, Esmon CT. et al. Endothelial cell protein C receptor acts as a cellular receptor for factor VIIa on endothelium. J Biol Chem 2007; 282: 11849-11857.
26 McLaughlin JN, Shen L, Holinstat M. et al. Functional selectivity of G protein signaling by agonist peptides and thrombin for the protease-activated receptor-1. J Biol Chem 2005; 280: 25048-25059.
27 Coughlin SR. Protease-activated receptors in hemostasis, thrombosis and vascular biology. J Thromb Haemost 2005; 03: 1800-1814.
28 Komarova YA, Mehta D, Malik AB. Dual regulation of endothelial junctional permeability. Sci STKE 2007; re8: 1-9.
29 Taylor Jr FB, Chang A, Hinshaw LB. et al. A model for thrombin protection against endotoxin. Thromb Res 1984; 36: 177-185.
30 Majno G, Palade GE. Studies on inflammation. 1. The effect of histamine and serotonin on vascular permeability: An electron microscopic study. J. Biophys Biochem Cytol 1961; 11: 571-605.
31 Jenkins RG, Su X, Su G. et al. Ligation of protease-activated receptor 1 enhances α_vβ₆ integrin-dependent TGF-β activation and promotes acute lung injury. J Clin Invest 2006; 116: 1606-1614.
32 Jesmin S, Gando S, Zaedi S. et al. Differential expression, time course and distribution of four PA Rs in rats with endotoxin-induced acute lung injury. Inflammation 2007; 30: 14-27.
33 Vogel SM, Gao X, Mehta D. et al. Abrogation of thrombin-induced increase in pulmonary microvascular permeability in PAR-1 knockout mice. Physiol Genomics 04: 137-145.
34 McLaughlin JN, Patterson MM, Malik AB. Protease-activated receptor-3 (PAR3) regulates PAR1 signaling by receptor dimerization. Proc Natl Acad Sci USA 2007; 104: 5662-5667.
35 Esmon CT. Role of coagulation inhibitors in inflammation. Thromb Haemost 2001; 86: 51-56.