Molecular basis of antithrombin deficiency

Beate Luxembourg; Daniel Delev; Christof Geisen; Michael Spannagl; Manuela Krause; Wolfgang Miesbach; Christine Heller; Frauke Bergmann; Ursula Schmeink; Ralf Grossmann; Edelgard Lindhoff-Last; Erhard Seifried; Johannes Oldenburg; Anna Pavlova

doi:10.1160/TH10-08-0538

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2011; 105(04): 635-646
DOI: 10.1160/TH10-08-0538

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Molecular basis of antithrombin deficiency

Beate Luxembourg

¹Institute of Transfusion Medicine and Immunohaematology, DRK Blood Donor Service Baden-Württemberg-Hessen, Frankfurt, Germany

²Department of Internal Medicine, Division of Vascular Medicine and Haemostaseology, University Hospital Frankfurt, Germany

,

Daniel Delev

³Clinic and Policlinic for Neurosurgery, University Clinic Bonn, Bonn, Germany

,

Christof Geisen

²Department of Internal Medicine, Division of Vascular Medicine and Haemostaseology, University Hospital Frankfurt, Germany

,

Michael Spannagl

⁴Department of Transfusion Medicine and Haemostaseology, Ludwig Maximilians University, Munich, Germany

,

Manuela Krause

⁵Division of Haemostaseology and Angiology, DKD, Wiesbaden, Germany

,

Wolfgang Miesbach

⁶Haemophilia Centre, University Hospital Frankfurt, Germany

,

Christine Heller

⁷Department of Paediatric Medicine, University Hospital Germany

,

Frauke Bergmann

⁸MVZ Wagnerstibbe, Hannover, Germany

,

Ursula Schmeink

⁹Praxis für Gefäßchirurgie und Phlebologie, Aachen, Germany

,

Ralf Grossmann

¹⁰Gemeinschaftspraxis Laborärzte Schweinfurt, Schweinfurt, Germany

,

Edelgard Lindhoff-Last

²Department of Internal Medicine, Division of Vascular Medicine and Haemostaseology, University Hospital Frankfurt, Germany

,

Erhard Seifried

¹Institute of Transfusion Medicine and Immunohaematology, DRK Blood Donor Service Baden-Württemberg-Hessen, Frankfurt, Germany

,

Johannes Oldenburg

¹¹Institute of Experimental Haematology and Transfusion Medicine, University Clinic Bonn, Bonn, Germany

,

Anna Pavlova

¹¹Institute of Experimental Haematology and Transfusion Medicine, University Clinic Bonn, Bonn, Germany

› Author Affiliations

Abstract

Summary

Antithrombin (AT) is the most important physiological inhibitor of coagulation proteases. It is activated by glycosaminoglycans such as heparin. Hereditary antithrombin deficiency is a rare disease that is mainly associated with venous thromboembolism. So far, more than 200 different mutations in the antithrombin gene (SERPINC1) have been described. The aim of our study was to characterise the molecular background in a large cohort of patients with AT deficiency. Mutation analysis was performed by direct sequencing of SERPINC1 in 272 AT-deficient patients. Large deletions were identified by multiplex PCR coupled with liquid chromatography or multiplex ligation-dependent probe amplification (MLPA) analysis. To predict the effect of SERPINC1 sequence variations on the pathogenesis of AT deficiency, in silico assessments, multiple sequence alignment, and molecular graphic imaging were performed. The mutation profile consisted of 59% missense, 10% non-sense, 8% splice site mutations, 15% small deletions/insertions/duplications, and 8% large deletions. Altogether 87 different mutations, including 42 novel mutations (22 missense and 20 null mutations), were identified. Of the novel missense mutations, nine are suspected to impair the conformational changes that are needed for AT activation, two to affect the central reactive loop or the heparin binding site, and six to impair the structural integrity of the molecule. Despite the heterogeneous background of AT deficiency, 10 AT variants occurred in multiple index patients. Characterisation of the SERPINC1 mutation profile in large cohorts of patients may help to further elucidate the pathogenesis of AT deficiency and to establish genotype-phenotype associations.

Keywords

Antithrombin deficiency - mutation analysis - DHPLC - thromboembolism

Full Text

References

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