Dok-1 negatively regulates platelet integrin αIIbβ3 outside-in signalling and inhibits thrombosis in mice

Masaru Niki; Manasa K. Nayak; Hong Jin; Neha Bhasin; Edward F. Plow; Pier Paolo Pandolfi; Paul B. Rothman; Anil K. Chauhan; Steven R. Lentz

doi:10.1160/TH15-05-0373

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2016; 115(05): 969-978
DOI: 10.1160/TH15-05-0373

Cellular Haemostasis and Platelets

Schattauer GmbH

Dok-1 negatively regulates platelet integrin αIIbβ3 outside-in signalling and inhibits thrombosis in mice

Masaru Niki

¹Department of Internal Medicine, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA

,

Manasa K. Nayak

²Department of Pediatrics, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA

,

Hong Jin^*

¹Department of Internal Medicine, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA

,

Neha Bhasin

²Department of Pediatrics, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA

,

Edward F. Plow

³Joseph J. Jacobs Center for Thrombosis and Vascular Biology, Department of Molecular Cardiology, Cleveland Clinic, Cleveland, Ohio, USA

,

Pier Paolo Pandolfi

⁴Cancer Research Institute, Beth Israel Deaconess Cancer Center, Departments of Medicine and Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

,

Paul B. Rothman

⁵Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

,

Anil K. Chauhan

¹Department of Internal Medicine, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA

,

Steven R. Lentz

¹Department of Internal Medicine, The University of Iowa Carver College of Medicine, Iowa City, Iowa, USA

› Author Affiliations

Abstract

Summary

Adaptor proteins play a critical role in the assembly of signalling complexes after engagement of platelet receptors by agonists such as collagen, ADP and thrombin. Recently, using proteomics, the Dok (downstream of tyrosine kinase) adapter proteins were identified in human and mouse platelets. In vitro studies suggest that Dok-1 binds to platelet integrin β3, but the underlying effects of Dok-1 on αIIbβ3 signalling, platelet activation and thrombosis remain to be elucidated. In the present study, using Dok-1-deficient (Dok-1^-/-) mice, we determined the phenotypic role of Dok-1 in αIIbβ3 signalling. We found that platelets from Dok-1^-/- mice displayed normal aggregation, activation of αIIβ3 (assessed by binding of JON/A), P-selectin surface expression (assessed by anti-CD62P), and soluble fibrinogen binding. These findings indicate that Dok-1 does not affect “inside-out” platelet signalling. Compared with platelets from wild-type (WT) mice, platelets from Dok-1^-/- mice exhibited increased clot retraction (p < 0.05 vs WT), increased PLCγ2 phosphorylation, and enhanced spreading on fibrinogen after thrombin stimulation (p < 0.01 vs WT), demonstrating that Dok-1 negatively regulates αIIbβ3 “outside-in” signalling. Finally, we found that Dok-1^-/- mice exhibited significantly shortened bleeding times and accelerated carotid artery thrombosis in response to photochemical injury (p < 0.05 vs WT mice). We conclude that Dok-1 modulates thrombosis and haemostasis by negatively regulating αIIbβ3 outside-in signalling.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Fibrinogen - haemostasis - integrin - platelets - thrombin - thrombosis

Full Text

References

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