Thromb Haemost 2004; 92(01): 36-41
DOI: 10.1160/TH04-02-0084
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Recombinant expression of mutations causing von Willebrand disease type Normandy: characterization of a combined defect of factor VIII binding and multimerization

Reinhard Schneppenheim
1   Paediatric Haematology & Oncology, University Children’s Hospital, Hamburg, Germany
,
Harald Lenk
2   University Children’s Hospital, Leipzig, Germany
,
Tobias Obser
1   Paediatric Haematology & Oncology, University Children’s Hospital, Hamburg, Germany
,
Johannes Oldenburg
3   Institute of Transfusion Medicine and Immune Haematology of the DRK Blood Donor Service, University of Frankfurt, Frankfurt, Germany
,
Florian Oyen
1   Paediatric Haematology & Oncology, University Children’s Hospital, Hamburg, Germany
,
Sonja Schneppenheim
1   Paediatric Haematology & Oncology, University Children’s Hospital, Hamburg, Germany
,
Rainer Schwaab
4   Institute of Experimental Hematology and Transfusion Medicine, University of Bonn, Bonn, Germany
,
Kerstin Will
5   Lab Association Prof. Arndt & Partners, Coagulation Lab, Hamburg, Germany
,
Ulrich Budde
5   Lab Association Prof. Arndt & Partners, Coagulation Lab, Hamburg, Germany
› Institutsangaben
Financial support: We gratefully acknowledge continuous financial support by the Deutsche Forschungsgemeinschaft (DFG grants Schn 325/4-1, Schn 325/4-2).
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Publikationsverlauf

Received 10. Februar 2004

Accepted 10. März 2004

Publikationsdatum:
29. November 2017 (online)

Summary

Von Willebrand disease type Normandy (VWD 2N) is caused by mutations at the factor VIII (FVIII) binding site of VWF, located at the amino-terminus of mature VWF. It is inherited in a recessive fashion and both homozygous and compound heterozygous mutations have been identified. Homozygous mutations are correlated with a clinical phenotype indistinguishable from mild hemophilia A by conventional laboratory tests, whereas compound heterozygosity with a quantitative defect may appear as VWD type 1 (VWD1). We have now identified and expressed a novel heterozygous mutation (Y795C) which is responsible for both, a defective FVIII-binding and aberrant multimers in a female patient with mild FVIII deficiency. Additionally we expressed another mutation (E787K), previously identified by us in a male patient with a severe ‘pseudohemophilic’ phenotype. Analysis of the FVIII binding and the multimer structure of the respective recombinant VWF mutants reproduced the observed phenotype: the FVIII binding defect in addition to the aberrant multimer structure of the patient with Y795C and the FVIII binding defect only, in the patient with E787K. Our results demonstrate the causative nature of the two mutations and emphasize the impact of ‘cysteine mutations’ on the multimer structure of VWF.

 
  • References

  • 1 Nishino M, Girma JP, Rothschild C. et al. New variant of von Willebrand disease with defective binding to factor VIII. Blood 1989; 74: 1591-9.
  • 2 Mazurier C, Dieval J, Jorieux S. et al. A new von Willebrand factor (vWF) defect in a patient with factor VIII (FVIII) deficiency but with normal levels and multimeric patterns of both plasma and platelet vWF. Characterization of abnormal vWF/FVIII interaction. Blood 1990; 75: 20-6.
  • 3 von Willebrand EA. Hereditär Pseudohemofili. Finska Läk Sellsk Handl 1926; 68: 87-112.
  • 4 Gaucher C, Mercier B, Jorieux S. et al. Identification of two point mutations in the von Willebrand factor gene of three families with the ‘Normandy’ variant of von Willebrand disease. Br J Haematol 1991; 78: 506-14.
  • 5 Schneppenheim R, Budde U, Krey S. et al. Results of a screening for von Willebrand disease type 2N in patients with suspected haemophilia A or von Willebrand disease type 1. Thromb Haemost 1996; 76: 598-602.
  • 6 Casonato A, Gaucher C, Pontara E. et al. Type 2N von Willebrand disease due to Arg91Gln substitution and a cytosine deletion in exon 18 of the von Willebrand factor gene. Br J Haematol 1998; 103: 39-41.
  • 7 Ginsburg D, Sadler JE. von Willebrand disease: a database of point mutations, insertions, and deletions. For the Consortium on von Willebrand Factor Mutations and Polymorphisms, and the Subcommittee on von Willebrand Factor of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis. Thromb Haemost 1993; 69: 177-84.
  • 8 Gaucher C, Dieval J, Mazurier C. Characterization of von Willebrand factor gene defects in two unrelated patients with type IIC von Willebrand disease. Blood 1994; 84: 1024-30.
  • 9 Schneppenheim R, Brassard J, Krey S. et al. Defective dimerisation of von Willebrand factor subunits due to a Cys–Arg mutation in type IID von Willebrand disease. Proc Natl Acad Sci USA 1996; 93: 3581-6.
  • 10 Jorieux S, Fressinaud E, Goudemand J. et al. Conformational changes in the D’ domain of von Willebrand factor induced by CYS 25 and CYS 95 mutations lead to factor VIII binding defect and multimeric impairment. Blood 2000; 95: 3139-45.
  • 11 Allen S, Abuzenadah AM, Blagg JL. et al. Two novel type 2N von Willebrand diseasecausing mutations that result in defective factor VIII binding, multimerization, and secretion of von Willebrand factor. Blood 2000; 95: 2000-7.
  • 12 Hardisty RM, McPherson JC. A one-stage factor VIII (antihaemophilic globulin) assay and its use on venous and capillary plasma. Thrombos Diathes Haemorrh 1962; 07: 215-29.
  • 13 Mazurier C, Parquet AGernez, Goudemand M. Enzyme-linked immunoabsorbent assay of factor VIII-related antigen. Interest in study of Von Willebrand’s disease. Pathol Biol Paris 1977; 25: 18-24.
  • 14 Macfarlane DE, Stibbe J, Kirby EP. et al. A method for assaying von Willebrand factor (ristocetin cofactor). Thromb Diath Haemorrh 1975; 34: 306-8.
  • 15 Brown JE, Bosak JO. An ELISA test for the binding of von Willebrand antigen to collagen. Thromb Res 1986; 43: 303-11.
  • 16 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor VIII/von Willebrand factor. Blood 1981; 57: 1140-3.
  • 17 Schneppenheim R, Plendl H, Budde U. Luminography an alternative assay for detection of von Willebrand factor multimers. Thromb Haemostas 1988; 60: 133-6.
  • 18 Goodeve AC, Eikenboom JC, Ginsburg D. et al. ISTH SSC Subcommittee on von Willebrand factor. A standard nomenclature for von Willebrand factor gene mutations and polymorphisms. On behalf of the ISTH SSC Subcommittee on von Willebrand factor. Thromb Haemost 2001; 85: 929-31.
  • 19 Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A laboratory manual. Cold Spring Harbor Lab. Press; 1989
  • 20 Saiki RK, Gelfand DH, Stoffel S. et al. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 1988; 239: 487-91.
  • 21 Bonthron DT, Handin RI, Kaufman RJ. et al. Structure of pre-pro-von Willebrand factor and its expression in heterologous cells. Nature 1986; 324: 270-3.
  • 22 Gaucher C, Hanss M, Dechavanne M. et al. Substitution of cysteine for phenylalanine 751 in mature von Willebrand factor is a novel candidate mutation in a family with type IIA von Willebrand disease. Br J Haematol 1993; 83: 94-9.
  • 23 Inbal A, Kornbrot N, Harrison P. et al. Effect of type IIB von Willebrand disease mutation Arg(545)Cys on platelet glycoprotein Ib binding-studies with recombinant von Willebrand factor. Thromb Haemost 1993; 70: 1058-62.
  • 24 Hilbert L, Gaucher C, Mazurier C. Identification of two mutations (Arg611Cys and Arg611His) in the A1 loop of von Willebrand factor (vWF) responsible for type 2 von Willebrand disease with decreased plateletdependent function of vWF. Blood 1995; 86: 1010-8.
  • 25 Siguret V, Ribba AS, Christophe O. et al. Characterization of recombinant von Willebrand factors mutated on cysteine 509 or 695. Thromb Haemost 1996; 76: 453-9.
  • 26 Eikenboom JC, Matsushita T, Reitsma PH. et al. Dominant type 1 von Willebrand disease caused by mutated cysteine residues in the D3 domain of von Willebrand factor. Blood 1996; 88: 2433-41.
  • 27 Leyte A, Voorberg J, Van Schijndel HB. et al. The pro-polypeptide of von Willebrand factor is required for the formation of a functional factor VIII-binding site on mature von Willebrand factor. Biochem J 1991; Feb 15; 274 Pt: (01) 257-61.
  • 28 Mayadas TN, Wagner DD. Vicinal cysteines in the prosequence play a role in von Willebrand factor multimer assembly. Proc Natl Acad Sci U S A 1992; 89: 3531-5.