Background von Willebrand disease (VWD) is a genetic bleeding disorder caused by defects of von Willebrand factor (VWF), quantitative (type 1 and 3) or qualitative (type 2). The laboratory phenotyping is heterogenic making diagnosis difficult. Objectives Complete laboratory analysis of VWD as an expansion of the previously reported cross-sectional family-based VWD study in the Czech Republic (BRNO-VWD) and Slovakia (BRA-VWD) under the name "Heart of Europe," in order to improve the understanding of laboratory phenotype/genotype correlation. Patients and Methods In total, 227 suspected VWD patients were identified from historical records. Complete laboratory analysis was established using all available assays, including VWF multimers and genetic analysis. Results A total of 191 patients (from 119 families) were confirmed as having VWD. The majority was characterized as a type 1 VWD, followed by type 2. Multimeric patterns concordant with laboratory phenotypes were found in approximately 83% of all cases. A phenotype/genotype correlation was present in 84% (77% type 1, 99% type 2, and 61% type 3) of all patients. Another 45 candidate mutations (23 novel variations), not found in the initial study, could be identified (missense 75% and truncating 24%). An exon 1-3 gene deletion was identified in 14 patients where no mutation was found by direct DNA sequencing, increasing the linkage up to 92%, overall. Conclusion This study provides a cross-sectional overview of the VWD population in a part of Central Europe. It is an addition to the previously published BRNO-VWD study, and provides important data to the International Society of Thrombosis and Haemostasis/European Association for Haemophilia and Allied Disorders VWD mutation database with identification of novel causal mutations.

Antwerp University Antwerp Belgium

Blood Coagulation and Vascular Medicine Center Goodheart Institute and Foundation in Nature Medicine Rotterdam The Netherlands

Department of Clinical Haematology University Hospital Brno Brno Czech Republic

Department of Haematology Antwerp University Hospital Edegem Belgium

Department of Haematology University F D Roosevelt Hospital Banská Bystrica Slovakia

Department of Laboratory Methods Faculty of Medicine Masaryk University Brno Czech Republic

Department of Pediatric Haematology University Hospital Brno Brno Czech Republic

Haemostasis Unit Antwerp University Hospital Edegem Belgium

Medicine and Health Sciences Haemostasis Research Unit Antwerp University Antwerp Belgium

National Hemophilia Center Department of Haematology and Blood Transfusion of the Medical School of the Comenius University Bratislava Slovakia

See more in PubMed

Rodeghiero F, Castaman G, Dini E. Epidemiological investigation of the prevalence of von Willebrand's disease. Blood. 1987;69(02):454–459. PubMed

Zhou Y F, Eng E T, Zhu J, Lu C, Walz T, Springer T A. Sequence and structure relationships within von Willebrand factor. Blood. 2012;120(02):449–458. PubMed PMC

Nyman D. Interaction of collagen with the factor VIII antigen-activity - von Willebrand factor complex. Thromb Res. 1977;11(03):433–438. PubMed

Goto S, Salomon D R, Ikeda Y, Ruggeri Z M. Characterization of the unique mechanism mediating the shear-dependent binding of soluble von Willebrand factor to platelets. J Biol Chem. 1995;270(40):23352–23361. PubMed

Timmons S, Kloczewiak M, Hawiger J. ADP-dependent common receptor mechanism for binding of von Willebrand factor and fibrinogen to human platelets. Proc Natl Acad Sci U S A. 1984;81(15):4935–4939. PubMed PMC

Working Party on von Willebrand Disease Classification . Sadler J E, Budde U, Eikenboom J C. Update on the pathophysiology and classification of von Willebrand disease: a report of the Subcommittee on von Willebrand Factor. J Thromb Haemost. 2006;4(10):2103–2114. PubMed

Tosetto A, Castaman G. How I treat type 2 variant forms of von Willebrand disease. Blood. 2015;125(06):907–914. PubMed

Schneppenheim R, Budde U.von Willebrand factor: the complex molecular genetics of a multidomain and multifunctional protein J Thromb Haemost 20119(Suppl 1):209–215. PubMed

Lillicrap D.Von Willebrand disease - phenotype versus genotype: deficiency versus disease Thromb Res 2007120(Suppl 1):S11–S16. PubMed

Jacobi P M, Gill J C, Flood V H, Jakab D A, Friedman K D, Haberichter S L. Intersection of mechanisms of type 2A VWD through defects in VWF multimerization, secretion, ADAMTS-13 susceptibility, and regulated storage. Blood. 2012;119(19):4543–4553. PubMed PMC

Federici A B.Classification of inherited von Willebrand disease and implications in clinical practice Thromb Res 2009124(Suppl 1):S2–S6. PubMed

Goodeve A C. The genetic basis of von Willebrand disease. Blood Rev. 2010;24(03):123–134. PubMed

Ng C, Motto D G, Di Paola J. Diagnostic approach to von Willebrand disease. Blood. 2015;125(13):2029–2037. PubMed PMC

Vangenechten I, Mayger K, Smejkal P. A comparative analysis of different automated von Willebrand factor glycoprotein Ib-binding activity assays in well typed von Willebrand disease patients. J Thromb Haemost. 2018;16(07):1268–1277. PubMed

Goodeve A, Eikenboom J, Castaman G. Phenotype and genotype of a cohort of families historically diagnosed with type 1 von Willebrand disease in the European study, Molecular and Clinical Markers for the Diagnosis and Management of Type 1 von Willebrand Disease (MCMDM-1VWD) Blood. 2007;109(01):112–121. PubMed

French Reference Center for von Willebrand disease . Veyradier A, Boisseau P, Fressinaud E. A laboratory phenotype/genotype correlation of 1167 French patients from 670 families with von willebrand disease: a new epidemiologic picture. Medicine (Baltimore) 2016;95(11):e3038. PubMed PMC

James P D, Notley C, Hegadorn C. The mutational spectrum of type 1 von Willebrand disease: results from a Canadian cohort study. Blood. 2007;109(01):145–154. PubMed

Vangenechten I, Smejkal P, Zapletal O. Analysis of von Willebrand disease in the South Moravian population (Czech Republic): results from the BRNO-VWD study. Thromb Haemost. 2019;119(04):594–605. PubMed

Favaloro E J, Oliver S. Evaluation of a new commercial von Willebrand factor multimer assay. Haemophilia. 2017;23(04):e373–e377. PubMed

Oliver S, Lau K KE, Chapman K, Favaloro E J. Laboratory testing for Von Willebrand factor multimers. Methods Mol Biol. 2017;1646:495–511. PubMed

Bowyer A E, Goodfellow K J, Seidel H. Evaluation of a semi-automated von Willebrand factor multimer assay, the Hydragel 5 von Willebrand multimer, by two European centers. Res Pract Thromb Haemost. 2018;2(04):790–799. PubMed PMC

Crist R A, Heikal N M, Rodgers G M, Grenache D G, Smock K J. Evaluation of a new commercial method for von Willebrand factor multimeric analysis. Int J Lab Hematol. 2018;40(05):586–591. PubMed

Pikta M, Zemtsovskaja G, Bautista H. Preclinical evaluation of a semi-automated and rapid commercial electrophoresis assay for von Willebrand factor multimers. J Clin Lab Anal. 2018;32(06):e22416. PubMed PMC

Oliver S, Vanniasinkam T, Mohammed S, Vong R, Favaloro E J. Semi-automated von Willebrand factor multimer assay for von Willebrand disease: further validation, benefits and limitations. Int J Lab Hematol. 2019;41(06):762–771. PubMed

Vangenechten I, Gadisseur A. Improving diagnosis of von Willebrand disease: reference ranges for von Willebrand factor multimer distribution. Res Pract Thromb Haemost. 2020;4(06):1024–1034. PubMed PMC

Raimondi D, Tanyalcin I, Ferté J.DEOGEN2: prediction and interactive visualization of single amino acid variant deleteriousness in human proteins Nucleic Acids Res 201745(W1):W201–W206. PubMed PMC

Ramensky V, Bork P, Sunyaev S. Human non-synonymous SNPs: server and survey. Nucleic Acids Res. 2002;30(17):3894–3900. PubMed PMC

Adzhubei I A, Schmidt S, Peshkin L. A method and server for predicting damaging missense mutations. Nat Methods. 2010;7(04):248–249. PubMed PMC

Adzhubei I, Jordan D M, Sunyaev S R. Predicting functional effect of human missense mutations using PolyPhen-2. Curr Protoc Hum Genet. 2013;Chapter 7:20. PubMed PMC

Schwarz J M, Cooper D N, Schuelke M, Seelow D. MutationTaster2: mutation prediction for the deep-sequencing age. Nat Methods. 2014;11(04):361–362. PubMed

James P D, Connell N T, Ameer B. ASH ISTH NHF WFH 2021 guidelines on the diagnosis of von Willebrand disease. Blood Adv. 2021;5(01):280–300. PubMed PMC

Schneppenheim R, Michiels J J, Obser T. A cluster of mutations in the D3 domain of von Willebrand factor correlates with a distinct subgroup of von Willebrand disease: type 2A/IIE. Blood. 2010;115(23):4894–4901. PubMed

Schneppenheim R, Budde U, Ruggeri Z M. A molecular approach to the classification of von Willebrand disease. Best Pract Res Clin Haematol. 2001;14(02):281–298. PubMed

Schneppenheim R, Budde U. Phenotypic and genotypic diagnosis of von Willebrand disease: a 2004 update. Semin Hematol. 2005;42(01):15–28. PubMed

Michiels J J, Berneman Z, Gadisseur A. Classification and characterization of hereditary types 2A, 2B, 2C, 2D, 2E, 2M, 2N, and 2U (unclassifiable) von Willebrand disease. Clin Appl Thromb Hemost. 2006;12(04):397–420. PubMed

Casonato A, Daidone V, Galletta E, Bertomoro A. Type 2B von Willebrand disease with or without large multimers: a distinction of the two sides of the disorder is long overdue. PLoS One. 2017;12(06):e0179566. PubMed PMC

Weiss H J, Sussman I I. A new von Willebrand variant (type I, New York): increased ristocetin-induced platelet aggregation and plasma von Willebrand factor containing the full range of multimers. Blood. 1986;68(01):149–156. PubMed

Holmberg L, Dent J A, Schneppenheim R, Budde U, Ware J, Ruggeri Z M. von Willebrand factor mutation enhancing interaction with platelets in patients with normal multimeric structure. J Clin Invest. 1993;91(05):2169–2177. PubMed PMC

Meyer D, Fressinaud E, Hilbert L, Ribba A S, Lavergne J M, Mazurier C. Type 2 von Willebrand disease causing defective von Willebrand factor-dependent platelet function. Best Pract Res Clin Haematol. 2001;14(02):349–364. PubMed

Rabinowitz I, Tuley E A, Mancuso D J. von Willebrand disease type B: a missense mutation selectively abolishes ristocetin-induced von Willebrand factor binding to platelet glycoprotein Ib. Proc Natl Acad Sci U S A. 1992;89(20):9846–9849. PubMed PMC

Ribba A S, Loisel I, Lavergne J M. Ser968Thr mutation within the A3 domain of von Willebrand factor (VWF) in two related patients leads to a defective binding of VWF to collagen. Thromb Haemost. 2001;86(03):848–854. PubMed

Flood V H, Lederman C A, Wren J S. Absent collagen binding in a VWF A3 domain mutant: utility of the VWF:CB in diagnosis of VWD. J Thromb Haemost. 2010;8(06):1431–1433. PubMed PMC

Castaman G, Federici A B, Tosetto A. Different bleeding risk in type 2A and 2M von Willebrand disease: a 2-year prospective study in 107 patients. J Thromb Haemost. 2012;10(04):632–638. PubMed

Favaloro E J, Pasalic L, Curnow J. Type 2M and Type 2A von Willebrand disease: similar but different. Semin Thromb Hemost. 2016;42(05):483–497. PubMed

Federici A B, Bucciarelli P, Castaman G. Management of inherited von Willebrand disease in Italy: results from the retrospective study on 1234 patients. Semin Thromb Hemost. 2011;37(05):511–521. PubMed

Mohl A, Marschalek R, Masszi T. An Alu-mediated novel large deletion is the most frequent cause of type 3 von Willebrand disease in Hungary. J Thromb Haemost. 2008;6(10):1729–1735. PubMed

Eikenboom J, Van Marion V, Putter H. Linkage analysis in families diagnosed with type 1 von Willebrand disease in the European study, molecular and clinical markers for the diagnosis and management of type 1 VWD. J Thromb Haemost. 2006;4(04):774–782. PubMed

Flood V H, Christopherson P A, Gill J C. Clinical and laboratory variability in a cohort of patients diagnosed with type 1 VWD in the United States. Blood. 2016;127(20):2481–2488. PubMed PMC

UK Haemophilia Centre Doctors' Organisation . Cumming A, Grundy P, Keeney S. An investigation of the von Willebrand factor genotype in UK patients diagnosed to have type 1 von Willebrand disease. Thromb Haemost. 2006;96(05):630–641. PubMed

Yadegari H, Driesen J, Pavlova A, Biswas A, Hertfelder H J, Oldenburg J. Mutation distribution in the von Willebrand factor gene related to the different von Willebrand disease (VWD) types in a cohort of VWD patients. Thromb Haemost. 2012;108(04):662–671. PubMed

Wellcome Trust Case Control Consortium . Smith N L, Chen M H, Dehghan A. Novel associations of multiple genetic loci with plasma levels of factor VII, factor VIII, and von Willebrand factor: the CHARGE (Cohorts for Heart and Aging Research in Genome Epidemiology) consortium. Circulation. 2010;121(12):1382–1392. PubMed PMC

van Loon J, Dehghan A, Weihong T. Genome-wide association studies identify genetic loci for low von Willebrand factor levels. Eur J Hum Genet. 2016;24(07):1096. PubMed PMC