A novel thromboxane A2 receptor N42S variant results in reduced surface expression and platelet dysfunction

Shaista P. Nisar; Marie Lordkipanidzé; Matthew L. Jones; Ban B. Dawood; Sherina Murden; Margaret R. Cunningham; Andrew D. Mumford; Jonathan T. Wilde; Steve P. Watson; Stuart J. Mundell; Gillian C. Lowe; the UK GAPP Study Group; on behalf of

doi:10.1160/TH13-08-0672

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2014; 111(05): 923-932
DOI: 10.1160/TH13-08-0672

Platelets and Blood Cells

Schattauer GmbH

A novel thromboxane A2 receptor N42S variant results in reduced surface expression and platelet dysfunction

Shaista P. Nisar

¹School of Physiology and Pharmacology, Medical Sciences Building, University of Bristol, Bristol, UK

,

Marie Lordkipanidzé

²Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, UK

,

Matthew L. Jones

³Bristol Heart Institute & School of Cellular and Molecular Medicine, University of Bristol, Bristol, Bristol, UK; 4West Midlands Adult Haemophilia Comprehensive Care Centre, University Hospital Brimingham, Edgbaston, Birmingham, UK

,

Ban B. Dawood

²Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, UK

,

Sherina Murden

³Bristol Heart Institute & School of Cellular and Molecular Medicine, University of Bristol, Bristol, Bristol, UK; 4West Midlands Adult Haemophilia Comprehensive Care Centre, University Hospital Brimingham, Edgbaston, Birmingham, UK

,

Margaret R. Cunningham

¹School of Physiology and Pharmacology, Medical Sciences Building, University of Bristol, Bristol, UK

,

Andrew D. Mumford

³Bristol Heart Institute & School of Cellular and Molecular Medicine, University of Bristol, Bristol, Bristol, UK; 4West Midlands Adult Haemophilia Comprehensive Care Centre, University Hospital Brimingham, Edgbaston, Birmingham, UK

,

Jonathan T. Wilde

⁴West Midlands Adult Haemophilia Comprehensive Care Centre, University Hospital Brimingham, Edgbaston, Birmingham, UK

,

Steve P. Watson

²Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, UK

,

Stuart J. Mundell

¹School of Physiology and Pharmacology, Medical Sciences Building, University of Bristol, Bristol, UK

,

Gillian C. Lowe

²Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, UK

,

the UK GAPP Study Group

› Institutsangaben

Abstract

Summary

A small number of thromboxane receptor variants have been described in patients with a bleeding history that result in platelet dysfunction. We have identified a patient with a history of significant bleeding, who expresses a novel heterozygous thromboxane receptor variant that predicts an asparagine to serine substitution (N42S). This asparagine is conserved across all class A GPCRs, suggesting a vital role for receptor structure and function. We investigated the functional consequences of the TP receptor heterozygous N42S substitution by performing platelet function studies on platelet-rich plasma taken from the patient and healthy controls. We investigated the N42S mutation by expressing the wild-type (WT) and mutant receptor in human embryonic kidney (HEK) cells. Aggregation studies showed an ablation of arachidonic acid responses in the patient, whilst there was right-ward shift of the U46619 concentration response curve (CRC). Thromboxane generation was unaffected. Calcium mobilisation studies in cells lines showed a rightward shift of the U46619 CRC in N42S–expressing cells compared to WT. Radioligand binding studies revealed a reduction in BMax in platelets taken from the patient and in N42S–expressing cells, whilst cell studies confirmed poor surface expression. We have identified a novel thromboxane receptor variant, N42S, which results in platelet dysfunction due to reduced surface expression. It is associated with a significant bleeding history in the patient in whom it was identified. This is the first description of a naturally occurring variant that results in the substitution of this highly conserved residue and confirms the importance of this residue for correct GPCR function.

Keywords

Inherited / acquired platelet disorders - gene mutations - receptors - platelet pharmacology - platelet pathology / inherited - acquired

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Referenzen

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