Human megakaryocytes confer tissue factor to a subset of shed platelets to stimulate thrombin generation

Marta Brambilla; Laura Facchinetti; Paola Canzano; Laura Rossetti; Nicola Ferri; Alessandra Balduini; Vittorio Abbonante; Daniela Boselli; Luigi De Marco; Matteo N.D. Di Minno; Vincenzo Toschi; Alberto Corsini; Elena Tremoli; Marina Camera

doi:10.1160/TH14-10-0830

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2015; 114(03): 579-592
DOI: 10.1160/TH14-10-0830

Cellular Haemostasis and Platelets

Schattauer GmbH

Human megakaryocytes confer tissue factor to a subset of shed platelets to stimulate thrombin generation

Authors

Marta Brambilla^*

¹Centro Cardiologico Monzino IRCCS, Milan, Italy
Laura Facchinetti^*

¹Centro Cardiologico Monzino IRCCS, Milan, Italy
Paola Canzano

¹Centro Cardiologico Monzino IRCCS, Milan, Italy
Laura Rossetti

¹Centro Cardiologico Monzino IRCCS, Milan, Italy
Nicola Ferri

²Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan, Italy

³Multimedia IRCCS, Milan Italy
Alessandra Balduini

⁴Department of Molecular Medicine, University of Pavia, IRCCS Policlinico San Matteo Foundation, Pavia, Italy
Vittorio Abbonante

⁴Department of Molecular Medicine, University of Pavia, IRCCS Policlinico San Matteo Foundation, Pavia, Italy
Daniela Boselli

¹Centro Cardiologico Monzino IRCCS, Milan, Italy
Luigi De Marco

⁵Centro di Riferimento Oncologico, Aviano, Italy
Matteo N.D. Di Minno

⁶Department of Clinical and Experimental Medicine, Federico II University, Naples, Italy
Vincenzo Toschi

⁷Division of Haematology and Blood Transfusion and Thrombosis Centre, AO Ospedale San Carlo Borromeo, Milan, Italy
Alberto Corsini

²Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan, Italy

³Multimedia IRCCS, Milan Italy
Elena Tremoli

¹Centro Cardiologico Monzino IRCCS, Milan, Italy

²Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan, Italy
Marina Camera

¹Centro Cardiologico Monzino IRCCS, Milan, Italy

²Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan, Italy

Abstract

Summary

Tissue factor (TF), the main activator of the blood coagulation cascade, has been shown to be expressed by platelets. Despite the evidence that both megakaryocytes and platelets express TF mRNA, and that platelets can make de novo protein synthesis, the main mechanism thought to be responsible for the presence of TF within platelets is through the uptake of TF positive microparticles. In this study we assessed 1) whether human megakaryocytes synthesise TF and transfer it to platelets and 2) the contribution of platelet-TF to the platelet hemostatic capacity. In order to avoid the cross-talk with circulating microparticles, we took advantage from an in vitro cultured megakaryoblastic cell line (Meg-01) able to differentiate into megakaryocytes releasing platelet-like particles. We show that functionally active TF is expressed in human megakaryoblasts, increased in megakaryocytes, and is transferred to a subset of shed platelets where it contributes to clot formation. These data were all confirmed in human CD34^pos- derived megakaryocytes and in their released platelets. The effect of TF silencing in Meg-megakaryoblasts resulted in a significant reduction of TF expression in these cells and also in Meg-megakaryocytes and Meg-platelets. Moreover, the contribution of platelet-TF to the platelet hemostatic capacity was highlighted by the significant delay in the kinetic of thrombin formation observed in platelets released by TF-silenced megakaryocytes. These findings provide evidences that TF is an endogenously synthesised protein that characterises megakaryocyte maturation and that it is transferred to a subset of newly-released platelets where it is functionally active and able to trigger thrombin generation.

Keywords

Megakaryocytes - platelets - tissue factor - thrombin generation

Full Text

References

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