Use of Targeted High-Throughput Sequencing for Genetic Classification of Patients with Bleeding Diathesis and Suspected Platelet Disorder

Oliver Andres; Eva-Maria König; Karina Althaus; Tamam Bakchoul; Peter Bugert; Stefan Eber; Ralf Knöfler; Erdmute Kunstmann; Georgi Manukjan; Oliver Meyer; Gabriele Strauß; Werner Streif; Thomas Thiele; Verena Wiegering; Eva Klopocki; Harald Schulze; on Behalf of the THROMKIDplus Study Group of the Society of Paediatric Oncology Haematology (Gesellschaft für Pädiatrische Onkologie und Hämatologie, GPOH) and the Society of Thrombosis Haemostasis Research (Gesellschaft für Thrombose- und Hämostaseforschung, GTH)

doi:10.1055/s-0038-1676813

TH Open, Table of Contents

CC BY 4.0 · TH Open 2018; 02(04): e445-e454
DOI: 10.1055/s-0038-1676813

Original Article

Georg Thieme Verlag KG Stuttgart · New York

Use of Targeted High-Throughput Sequencing for Genetic Classification of Patients with Bleeding Diathesis and Suspected Platelet Disorder

Authors

Oliver Andres

¹University Children's Hospital, University of Würzburg, Würzburg, Germany
Eva-Maria König

²Institute of Human Genetics, University of Würzburg, Würzburg, Germany
Karina Althaus

³Centre for Clinical Transfusion Medicine, University Hospital of Tübingen, Tübingen, Germany

⁴Institute for Transfusion Medicine, University of Greifswald, Greifswald, Germany
Tamam Bakchoul

³Centre for Clinical Transfusion Medicine, University Hospital of Tübingen, Tübingen, Germany

⁴Institute for Transfusion Medicine, University of Greifswald, Greifswald, Germany
Peter Bugert

⁵DRK-Blutspendedienst Baden-Württemberg-Hessen, Institute for Transfusion Medicine and Immunology, Heidelberg University, Mannheim, Germany
Stefan Eber

⁶University Children's Hospital, Technical University Munich, Munich, Germany
Ralf Knöfler

⁷Department of Pediatrics, Carl Gustav Carus University Hospital, Dresden, Germany
Erdmute Kunstmann

²Institute of Human Genetics, University of Würzburg, Würzburg, Germany
Georgi Manukjan

⁸Institute of Experimental Biomedicine, University Hospital Würzburg, Würzburg, Germany
Oliver Meyer

⁹Institute for Transfusion Medicine, Charité—Universitätsmedizin Berlin, Berlin, Germany
Gabriele Strauß

¹⁰Department for Pediatric Oncology and Hematology, HELIOS Klinikum Berlin-Buch, Berlin, Germany
Werner Streif

¹¹Department of Pediatrics, Medical University Innsbruck, Innsbruck, Austria
Thomas Thiele

⁴Institute for Transfusion Medicine, University of Greifswald, Greifswald, Germany
Verena Wiegering

¹University Children's Hospital, University of Würzburg, Würzburg, Germany
Eva Klopocki

²Institute of Human Genetics, University of Würzburg, Würzburg, Germany
Harald Schulze

⁸Institute of Experimental Biomedicine, University Hospital Würzburg, Würzburg, Germany

on Behalf of the THROMKIDplus Study Group of the Society of Paediatric Oncology Haematology (Gesellschaft für Pädiatrische Onkologie und Hämatologie, GPOH) and the Society of Thrombosis Haemostasis Research (Gesellschaft für Thrombose- und Hämostaseforschung, GTH)

Abstract

Inherited platelet disorders (IPD) form a rare and heterogeneous disease entity that is present in about 8% of patients with non-acquired bleeding diathesis. Identification of the defective cellular pathway is an important criterion for stratifying the patient's individual risk profile and for choosing personalized therapeutic options. While costs of high-throughput sequencing technologies have rapidly declined over the last decade, molecular genetic diagnosis of bleeding and platelet disorders is getting more and more suitable within the diagnostic algorithms. In this study, we developed, verified, and evaluated a targeted, panel-based next-generation sequencing approach comprising 59 genes associated with IPD for a cohort of 38 patients with a history of recurrent bleeding episodes and functionally suspected, but so far genetically undefined IPD. DNA samples from five patients with genetically defined IPD with disease-causing variants in WAS, RBM8A, FERMT3, P2YR12, and MYH9 served as controls during the validation process. In 40% of 35 patients analyzed, we were able to finally detect 15 variants, eight of which were novel, in 11 genes, ACTN1, AP3B1, GFI1B, HPS1, HPS4, HPS6, MPL, MYH9, TBXA2R, TPM4, and TUBB1, and classified them according to current guidelines. Apart from seven variants of uncertain significance in 11% of patients, nine variants were classified as likely pathogenic or pathogenic providing a molecular diagnosis for 26% of patients. This report also emphasizes on potentials and pitfalls of this tool and prospectively proposes its rational implementation within the diagnostic algorithms of IPD.

Keywords

next-generation sequencing - molecular genetics - thrombocytopenia - thrombocytopathy - platelet function disorder

Full Text

References

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