Homozygous type 2N R854W von Willebrand factor is poorly secreted and causes a severe von Willebrand disease phenotype

被引:20
作者
Castaman, G. [1 ]
Giacomelli, S. H. [1 ]
Jacobi, P. [2 ,3 ]
Obser, T. [4 ]
Budde, U. [5 ]
Rodeghiero, F. [1 ]
Haberichter, S. L. [2 ,3 ]
Schneppenheim, R. [4 ]
机构
[1] San Bortolo Hosp, Dept Cellular Therapies & Haematol, I-36100 Vicenza, Italy
[2] Childrens Hosp Wisconsin, Childrens Res Inst, Milwaukee, WI 53201 USA
[3] Med Coll Wisconsin, Dept Pediat, Milwaukee, WI 53226 USA
[4] Univ Med Ctr, Dept Paediat Haematol & Oncol, Hamburg, Germany
[5] Asklepios Clin Altona, Medilys Lab, Hamburg, Germany
关键词
gene mutation; inherited bleeding disorders; von Willebrand disease; von Willebrand factor; FACTOR-VIII BINDING; VONWILLEBRAND DISEASE; MULTIMERIZATION; MUTATION; DIAGNOSIS; DOMAIN; STORAGE; PATHOPHYSIOLOGY; EXPRESSION; NORMANDY;
D O I
10.1111/j.1538-7836.2010.03971.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Background: von Willebrand disease (VWD) type Normandy (VWD 2N) is caused by mutations at the factor (F) VIII-binding site of von Willebrand factor (VWF), located in the D' and D3 domains on the N-terminus of mature VWF. The R854Q mutation is the most frequent cause of this phenotype. Objectives: We report the characterization of a homozygous VWD 2N mutation, R854W, detected in a patient with a severe VWD phenotype. Methods: The plasma VWF phenotype was studied, transient expression of recombinant mutant full-length VWF in 293 EBNA cells was performed, and the results were compared with those obtained with wild-type (WT) VWF. Furthermore, expression was also examined in HEK293 cells, which form Weibel-Palade body-like granules when transfected with WT VWF. Results: The multimer analysis of plasma VWF showed the lack of the typical triplet structure, with the presence of the central band only, and a relative decrease in the high molecular mass multimers. Homozygous expression of recombinant R854W VWF resulted in normal amounts of cellular VWF, but with a severe reduction in secretion into the medium. Severe reductions in FVIII binding to R854W VWF, glycoprotein Ib binding activity and collagen binding of secreted W854 VWF was observed, and reproduced the phenotypic parameters of plasma VWF. In HEK293 cells, homozygous R854W VWF failed to form Weibel-Palade body-like granules. Conclusions: Our results demonstrate that a homozygous R854W mutation in the D' domain of VWF induces impaired secretion and activity of the protein, thereby explaining the severe phenotype of the patient.
引用
收藏
页码:2011 / 2016
页数:6
相关论文
共 26 条
[1]   Two novel type 2N von Willebrand disease-causing mutations that result in defective factor VIII binding, multimerization, and secretion of von Willebrand factor [J].
Allen, S ;
Abuzenadah, AM ;
Blagg, JL ;
Hinks, J ;
Nesbitt, IM ;
Goodeve, AC ;
Gursel, T ;
Ingerslev, J ;
Peake, IR ;
Daly, ME .
BLOOD, 2000, 95 (06) :2000-2007
[2]   Shear-induced unfolding activates von Willebrand factor A2 domain for proteolysis [J].
Baldauf, C. ;
Schneppenheim, R. ;
Stacklies, W. ;
Obser, T. ;
Pieconka, A. ;
Schneppenheim, S. ;
Budde, U. ;
Zhou, J. ;
Graeter, F. .
JOURNAL OF THROMBOSIS AND HAEMOSTASIS, 2009, 7 (12) :2096-2105
[3]  
Bowen DJ, 1998, THROMB HAEMOSTASIS, V80, P32
[4]   AN ELISA TEST FOR THE BINDING OF VONWILLEBRAND ANTIGEN TO COLLAGEN [J].
BROWN, JE ;
BOSAK, JO .
THROMBOSIS RESEARCH, 1986, 43 (03) :303-311
[5]   Detailed von Willebrand factor multimer analysis in patients with von Willebrand disease in the European study, molecular and clinical markers for the diagnosis and management of type 1 von Willebrand disease (MCMDM-1VWD) [J].
Budde, U. ;
Schneppenheim, R. ;
Eikenboom, J. . ;
Goodeve, A. ;
Will, K. ;
Drewke, E. ;
Castaman, G. ;
Rodeghiero, F. ;
Federici, A. B. ;
Batlle, J. . ;
Perez, A. ;
Meyer, D. ;
Mazurier, C. ;
Goudemand, J. ;
Ingerslev, J. ;
Habart, D. ;
Vorlova, Z. ;
Holmberg, L. ;
Lethagen, S. ;
Pasi, J. ;
Hill, F. ;
Peake, I. .
JOURNAL OF THROMBOSIS AND HAEMOSTASIS, 2008, 6 (05) :762-771
[6]   Factor VIII accelerates proteolytic cleavage of von Willebrand factor by ADAMTS13 [J].
Cao, Wenjing ;
Krishnaswamy, Sriram ;
Camire, Rodney M. ;
Lenting, Peter J. ;
Zheng, X. Long .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (21) :7416-7421
[7]   FACTOR-VIII-C INCREASES AFTER DESMOPRESSIN IN A SUBGROUP OF PATIENTS WITH AUTOSOMAL RECESSIVE SEVERE VON-WILLEBRAND DISEASE [J].
CASTAMAN, G ;
LATTUADA, A ;
MANNUCCI, PM ;
RODEGHIERO, F .
BRITISH JOURNAL OF HAEMATOLOGY, 1995, 89 (01) :147-151
[8]   RECESSIVE INHERITANCE OF VONWILLEBRANDS DISEASE TYPE-I [J].
EIKENBOOM, JCJ ;
REITSMA, PH ;
PEERLINCK, KMJ ;
BRIET, E .
LANCET, 1993, 341 (8851) :982-986
[9]  
Federici AB, 2004, HAEMATOLOGICA, V89, P77
[10]   Clinical and molecular predictors of thrombocytopenia and risk of bleeding in patients with vonWillebrand disease type 2B: a cohort study of 67 patients [J].
Federici, Augusto B. ;
Mannucci, Pier M. ;
Castaman, Giancarlo ;
Baronciani, Luciano ;
Bucciarelli, Paolo ;
Canciani, Maria T. ;
Pecci, Alessandro ;
Lenting, Peter J. ;
De Groot, Philip G. .
BLOOD, 2009, 113 (03) :526-534