Thromb Haemost 2017; 117(03): 556-569
DOI: 10.1160/TH16-07-0515
Cellular Haemostasis and Platelets
Schattauer GmbH

Alterations of the platelet proteome in type I Glanzmann thrombasthenia caused by different homozygous delG frameshift mutations in ITGA2B

Stefan Loroch
1  Leibniz-Institut für Analytische Wissenschaften – ISAS – e. V., Dortmund, Germany
,
Katharina Trabold
2  Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany
,
Stepan Gambaryan
2  Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany
3  Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, S. Petersburg, Russia
,
Cora Reiß
2  Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany
,
Kathrin Schwierczek
2  Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany
,
Ingrid Fleming
4  Institute for Vascular Signalling, Centre for Molecular Medicine, Goethe-University, Frankfurt, Germany
5  German Centre for Cardiovascular Research (DZHK), RheinMain, Germany
,
Albert Sickmann
1  Leibniz-Institut für Analytische Wissenschaften – ISAS – e. V., Dortmund, Germany
6  Medizinisches Proteom Center, Medizin-ische Fakultät, Ruhr Universität Bochum, Bochum, Germany
7  Department of Chemistry, College of Physical Sciences, University of Aberdeen, Aberdeen, UK
,
Wolfgang Behnisch
8  Department of Pediatric Oncology, Hematology, Immunology and Pulmonology, University Medical Center, Heidelberg, Germany
,
Barbara Zieger
9  Department of Pediatrics and Adolescent Medicine, Division of Pediatric Hematology and Oncology, Medical Center – Faculty of Medicine, University of Freiburg, Germany
,
René P. Zahedi
1  Leibniz-Institut für Analytische Wissenschaften – ISAS – e. V., Dortmund, Germany
,
Ulrich Walter
2  Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany
5  German Centre for Cardiovascular Research (DZHK), RheinMain, Germany
,
Kerstin Jurk
2  Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany
5  German Centre for Cardiovascular Research (DZHK), RheinMain, Germany
› Author Affiliations
Further Information

Publication History

Received: 08 July 2016

Accepted after major revision: 09 January 2016

Publication Date:
22 November 2017 (online)

Summary

Glanzmann thrombasthenia (GT) is one of the best characterised inherited platelet function disorders but global platelet proteome has not been determined in these patients. We investigated the proteome and function of platelets from two patients with type I GT, caused by different homozygous ITGA2b mutations, from family members and unrelated controls. The global proteome of highly purified washed platelets was quantified by liquid chromatography-mass spectrometry (LC-MS) and targeted MS-methods. Platelet function was analysed by flow cytometry, light transmission aggregometry and flow-based as-says. Platelets from GT patients showed less than 5 % relative levels of the integrin subunit αIIb and 5–9 % fibrinogen compared to controls. These patients demonstrated loss of αIIbβ3-dependent platelet function, but normal platelet granule secretion induced by physiological agonists. Platelets from heterozygous family members of a patient expressed 50–60 % of control αIIb levels which were sufficient for normal αIIbβ3-dependent platelet function. Studying type I GT as model disease we established quantitative LC-MS to detect and clearly distinguish normal platelets, platelets from GT heterozygotes and platelets from GT patients. Diminished levels of factor XIIIB chain, plasminogen and carboxypeptidase 2B were identified in thrombasthenic platelets. Additionally, GT platelets showed up to 2.5-fold increased levels of FcγRIIA and laminin-α4 chain. Elevated levels of platelet FcγRIIA was associated with increased CD63-surface expression after FcγRIIA-crosslinking in one GT-patient which might present a compensatory mechanism of platelet activation in GT. We demonstrate that quantitative LC-MS based proteomics is suitable to validate known but also to identify previously unknown protein level changes of dysfunctional platelets.

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