CD44 sensitivity of platelet activation, membrane scrambling and adhesion under high arterial shear rates

Guilai Liu; Guoxing Liu; Kousi Alzoubi; Madhumita Chatterjee; Britta Walker; Patrick Münzer; Dong Luo; Anja T. Umbach; Bernat Elvira; Hong Chen; Jakob Voelkl; Michael Föller; Tak Z. Mak; Oliver Borst; Meinrad Gawaz; Florian Lang

doi:10.1160/TH14-10-0847

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2016; 115(01): 99-108
DOI: 10.1160/TH14-10-0847

Cellular Haemostasis and Platelets

Schattauer GmbH Schattauer

CD44 sensitivity of platelet activation, membrane scrambling and adhesion under high arterial shear rates

Guilai Liu^*

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Guoxing Liu^*

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Kousi Alzoubi

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Madhumita Chatterjee

²Department of Cardiology and Cardiovascular Medicine, University of Tübingen, Tübingen, Germany

,

Britta Walker

¹Department of Physiology, University of Tübingen, Tübingen, Germany

²Department of Cardiology and Cardiovascular Medicine, University of Tübingen, Tübingen, Germany

,

Patrick Münzer

¹Department of Physiology, University of Tübingen, Tübingen, Germany

²Department of Cardiology and Cardiovascular Medicine, University of Tübingen, Tübingen, Germany

,

Dong Luo

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Anja T. Umbach

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Bernat Elvira

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Hong Chen

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Jakob Voelkl

¹Department of Physiology, University of Tübingen, Tübingen, Germany

,

Michael Föller

¹Department of Physiology, University of Tübingen, Tübingen, Germany

³Campbell Family Institute for Breast Cancer Research, Ontario Cancer Institute, UHN, Toronto, Ontario, Canada

,

Tak Z. Mak

³Campbell Family Institute for Breast Cancer Research, Ontario Cancer Institute, UHN, Toronto, Ontario, Canada

,

Oliver Borst

¹Department of Physiology, University of Tübingen, Tübingen, Germany

²Department of Cardiology and Cardiovascular Medicine, University of Tübingen, Tübingen, Germany

,

Meinrad Gawaz

²Department of Cardiology and Cardiovascular Medicine, University of Tübingen, Tübingen, Germany

,

Florian Lang

¹Department of Physiology, University of Tübingen, Tübingen, Germany

› Author Affiliations

Abstract

Summary

CD44 is required for signalling of macrophage migration inhibitory factor (MIF), an anti-apoptotic pro-inflammatory cytokine. MIF is expressed and released from blood platelets, key players in the orchestration of occlusive vascular disease. Nothing is known about a role of CD44 in the regulation of platelet function. The present study thus explored whether CD44 modifies degranulation (P-selectin exposure), integrin activation, caspase activity, phosphatidylserine exposure on the platelet surface, platelet volume, Orai1 protein abundance and cytosolic Ca²⁺-activity ([Ca²⁺]i). Platelets from mice lacking CD44 (cd44^-/- ) were compared to platelets from corresponding wild-type mice (cd44^+/+ ). In resting platelets, P-selectin abundance, α_llbβ₃ inte-grin activation, caspase-3 activity and phosphatidylserine exposure were negligible in both genotypes and Orai1 protein abundance, [Ca²⁺]_i, and volume were similar in cd44^-/- and cd44^+/+ platelets. Platelet degranulation and α_llbβ₃ integrin activation were significantly increased by thrombin (0.02 U/ml), collagen related peptide (CRP, 2 µg/ml and Ca²⁺-store depletion with thapsigargin (1 µM), effects more pronounced in cd44^-/- than in cd44^+/+ platelets. Thrombin (0.02 U/ml) increased platelet [Ca²⁺]i, caspase-3 activity, phosphatidylserine exposure and Orai1 surface abundance, effects again significantly stronger in cd44^-/- than in cd44^+/+ platelets. Thrombin further decreased forward scatter in cd44^-/- and cd44^+/+ platelets, an effect which tended to be again more pronounced in cd44^-/- than in cd44^+/+ platelets. Platelet adhesion and in vitro thrombus formation under high arterial shear rates (1,700 s^-1) were significantly augmented in cd44^-/- mice. In conclusion, genetic deficiency of CD44 augments activation, apoptosis and prothrombotic potential of platelets.

Keywords

CD44 - platelet activation - cytosolic Ca²⁺ concentration - phosphatidylserine translocation - thrombus formation

Full Text

References

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