Mice Lacking PECAM-1 and Ceacam1 Have Enhanced Platelet Secretion and Thrombus Growth: Novel Link with PAR4

Fahd A. Kuriri; Genia Burchall; Fehaid Alanazi; Juliana Antonipillai; Gasim Dobie; Nicole Beachemin; Denise E. Jackson

doi:10.1055/a-1663-8108

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2022; 122(06): 961-973
DOI: 10.1055/a-1663-8108

Cellular Haemostasis and Platelets

Mice Lacking PECAM-1 and Ceacam1 Have Enhanced Platelet Secretion and Thrombus Growth: Novel Link with PAR4

Fahd A. Kuriri

¹Thrombosis and Vascular Diseases Laboratory, School of Health and Biomedical Sciences, STEM College, RMIT University, Bundoora, Victoria, Australia

²Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Shaqra University, Shaqra, Riyadh Province, Saudi Arabia

,

Genia Burchall

¹Thrombosis and Vascular Diseases Laboratory, School of Health and Biomedical Sciences, STEM College, RMIT University, Bundoora, Victoria, Australia

,

Fehaid Alanazi

¹Thrombosis and Vascular Diseases Laboratory, School of Health and Biomedical Sciences, STEM College, RMIT University, Bundoora, Victoria, Australia

³Department of Clinical Laboratory Sciences, College of Applied Medical Sciences-AlQurayyat, Jouf University, Saudi Arabia

,

Juliana Antonipillai

¹Thrombosis and Vascular Diseases Laboratory, School of Health and Biomedical Sciences, STEM College, RMIT University, Bundoora, Victoria, Australia

,

Gasim Dobie

⁴Department of Medical Laboratory Technology, Jazan University, Jazan, Saudi Arabia

,

Nicole Beachemin

⁵Department of Oncology, Medicine and Biochemistry, Goodman Cancer Research Centre, McGill University, Montreal, Canada

,

Denise E. Jackson

¹Thrombosis and Vascular Diseases Laboratory, School of Health and Biomedical Sciences, STEM College, RMIT University, Bundoora, Victoria, Australia

› Author Affiliations

Abstract

The Ig-ITIM bearing receptors, PECAM-1 and CEACAM1, have been shown net negative regulators of platelet–collagen interactions and hemiITAM signaling pathways. In this study, a double knockout (DKO) mouse was developed with deleted PECAM-1 and CEACAM1 to study their combined contribution in platelet activation by glycoprotein VI, C-type lectin-like receptor 2, protease activated receptor (PAR4), ADP purinergic receptors, and thromboxane receptor (TP) A2 pathways. In addition, their collective contribution was examined in thrombus formation under high shear and microvascular thrombosis using in vivo models. DKO platelets responded normally to ADP purinergic receptors and the TP A2 pathway. However, DKO platelets released significantly higher amounts of P-selectin compared with hyper-responsive Pecam-1^−/− or Ceacam1^−/− versus wild-type (WT) upon stimulation with collagen-related peptide or rhodocytin. In contrast, DKO platelets showed increased amounts of P-selectin exposure upon stimulation with PAR4 agonist peptide or thrombin but not Pecam-1^−/− , Ceacam1^−/− , or WT platelets. Blockade of phospholipase C (PLC) or Rho A kinase revealed that DKO platelets enhanced α-granule release via PAR4/Gαq/PLC signaling without crosstalk with Src/Syk or G_12/13 signaling pathways. Severely delayed clot retraction in vitro was observed in DKO phenotype. The DKO model revealed a significant increase in thrombus formation compared with the hyper-responsive Ceacam1^−/− or Pecam-1^−/− versus WT phenotype. DKO platelets have similar glycoprotein surface expression compared with Pecam-1^−/− , Ceacam1^−/− , and WT platelets. This study demonstrates that PECAM-1 and CEACAM1 work in concert to negatively regulate hemiITAM signaling, platelet–collagen interactions, and PAR4 Gαq protein- coupled signaling pathways. Both PECAM-1 and CEACAM1 are required for negative regulation of platelet activation and microvascular thrombosis in vivo.

Keywords

pecam-1 - ceacam1 - ITIM - PAR4 - GPCR signaling - platelets

Full Text

References

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