An Arg760Cys mutation in the consensus sequence of the von Willebrand factor propeptide cleavage site is responsible for a new von Willebrand disease variant

Casonato, Alessandra; Sartorello, Francesca; Cattini, Maria Grazia; Pontara, Elena; Soldera, Carmen; Bertomoro, Antonella; Girolami, Antonio

doi:10.1182/blood-2002-04-1046

Cited by 43 publications

(51 citation statements)

References 40 publications

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“…Mature VWF adopts a conformation for the N-terminal FVIII binding domain that supports optimal FVIII binding. VWF mutations that prevent propeptide cleavage result in impaired FVIII binding, [44][45][46] whereas VWF expressed without the propeptide exhibits a 6-fold lower affinity for FVIII in vitro than full-length, multimeric VWF. 47 Together, these reports suggest that the VWF propeptide mediates optimal folding of the D9D3 domains for FVIII binding.…”

Section: Discussionmentioning

confidence: 99%

A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

Yee

Gildersleeve

et al. 2014

Blood

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• The D9D3 domains of VWF are sufficient to stabilize FVIII in vivo.• The prolongation of VWF D9D3 survival in vivo by Fc fusion elevates FVIII levels in the setting of VWF but not FVIII deficiency.Plasma factor VIII (FVIII) and von Willebrand factor (VWF) circulate together as a complex. We identify VWF fragments sufficient for FVIII stabilization in vivo and show that hepatic expression of the VWF D9D3 domains (S764-P1247), either as a monomer or a dimer, is sufficient to raise FVIII levels in Vwf 2/2 mice from a baseline of ∼5% to 10%, to ∼50% to 100%. These results demonstrate that a fragment containing only ∼20% of the VWF sequence is sufficient to support FVIII stability in vivo. Expression of the VWF D9D3 fragment fused at its C terminus to the Fc segment of immunoglobulin G1 results in markedly enhanced survival in the circulation (t 1/2 > 7 days), concomitant with elevated plasma FVIII levels (>25% at 7 days) in Vwf 2/2 mice. Although the VWF D9D3-Fc chimera also exhibits markedly prolonged survival when transfused into FVIII-deficient mice, the cotransfused FVIII is rapidly cleared. Kinetic binding studies show that VWF propeptide processing of VWF D9D3 fragments is required for optimal FVIII affinity. The reduced affinity of VWF D9D3 and VWF D9D3-Fc for FVIII suggests that the shortened FVIII survival in FVIII-deficient mice transfused with FVIII and VWF D9D3/D9D3-Fc is due to ineffective competition of these fragments with endogenous VWF for FVIII binding. (Blood. 2014; 124(3):445-452)

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Section: Discussionmentioning

confidence: 99%

A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

Yee

Gildersleeve

et al. 2014

Blood

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“…19 Briefly, microtitration plates were coated with anti-VWF polyclonal antibody and left overnight at +4°C (Dako, Glostrup, Denmark Genetic analysis was performed on exons 17-20 of the VWF gene as previously described. 13 Amino acid residues were numbered from the ATG initiation codon (residue 1) of the pre-pro-VWF. The out of range values are highlighted in bold.…”

Section: Methodsmentioning

confidence: 99%

“…12 Three mutations (p.E787K, p.T791M and p.R816W) are associated with the most severe forms of type 2N VWD. 12 All the abovementioned mutations are located in the D' domain, but others have been reported in exon 17 13 or D3 domains. [14][15][16] Type 2N VWD patients are homozygotes or compound heterozygotes for different type 2N…”

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confidence: 99%

Type 2N von Willebrand disease: Characterization and diagnostic difficulties

et al. 2017

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Introduction An abnormal factor VIII (FVIII) binding capacity of von Willebrand factor (VWF) identifies type 2N von Willebrand disease (VWD). Type 2N VWD patients are identified by means of the VWF FVIII binding (VWF:FVIIIB) assay, and especially their VWF:FVIIIB/VWF:Ag ratio (VWF:FVIIIB ratio). Aim We report on our 15‐year experience of diagnosing type 2N VWD. Methods We have performed 2178 VWF:FVIIIB assays in bleeders and normal subjects. Results von Willebrand factor (VWF):FVIIIB was reduced in 682, but only 60 had low VWF:FVIIIB ratios (<0.74). Among nine patients who had a VWF:FVIIIB ratio below 0.3, four had normal VWF levels and were homozygotes for the p.R854Q mutation; the other five had low VWF levels due to a quantitative VWF mutation combined with p.R854Q. The VWF:FVIIIB ratio ranged between 0.3 and 0.73 in 51 subjects; 34 of them were heterozygotes for the p.R854Q mutation, while one carried the p.R760C. The heterozygotes for type 2N included subjects with or without bleeding symptoms, the former with significantly lower mean VWF levels than the latter. Among the 116 normal subjects tested, six were heterozygotes for the p.R854Q mutation (all asymptomatic). Conclusions The prevalence of type 2N in our VWD cohort was 2.5%, and 5.2% of the general population in Northeast Italy was found heterozygous for the p.R854Q mutation. It might be difficult to reveal a type 2N defect using routine tests alone, especially when it is combined with a quantitative VWF mutation. Accordingly, we always recommend VWF:FVIIIB assay in the diagnostic workup of VWD.

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“…The only exception was the R763C mutation in exon 17, which interferes with the cleavage of the VWF propeptide, the persistence of which sterically hinders factor VIII binding to the VWF monomer. 26 No Italian type 2N VWD patients carrying mutations other than R854Q mutation have been reported to date, prompting us to hypothesize a founder effect in our type 2N VWD patients. To test this hypothesis, 13 unrelated families with type 2N VWD were studied (including both affected and unaffected members) to seek a common haplotype associated with the disorder.…”

Section: Discussionmentioning

confidence: 99%

“…The first type 2N patient was found to carry the T791M VWF mutation, while a number of type 2N mutations were described in other patients, including C788Y, Y795C, C804F, R816W, R854Q, C858F D879N and R763C. [24][25][26] Despite the considerable number of type 2N mutations reported to date in different countries, the R854Q mutation was almost the only one to be found in our cohort of type 2N VWD patients, be they homozygotes, compound heterozygotes or heterozygotes with no bleeding symptoms. The only exception was the R763C mutation in exon 17, which interferes with the cleavage of the VWF propeptide, the persistence of which sterically hinders factor VIII binding to the VWF monomer.…”

Section: Discussionmentioning

confidence: 99%

A common ancestor more than 10,000 years old for patients with R854Q-related type 2N von Willebrand's disease in Italy

et al. 2012

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ARTICLES 147 HemostasisThe impaired capacity of von Willebrand factor to carry factor VIII is identified as type 2N von Willebrand's disease. R854Q is the most common type 2N mutation, and almost the only one identified in Italy. This aim of this study was to ascertain whether R854Q mutations in a cohort of Italian patients with type 2N von Willebrand's disease originated from a single event or recurrent events. Thirteen unrelated Italian families were investigated, analyzing the von Willebrand factor gene haplotype associated with the R854Q mutation. A common haplotype emerged in all the families, extending from single nucleotide polymorphisms rs2166902 to rs216293 over 48.2 kb and including five intragenic markers. This haplotype is infrequent in the healthy Italian population (17% versus 100%, P<0.0001) and each genetic marker within the said haplotype is similarly rare. These data strongly suggest a founder effect, with a single R854Q mutation event being the cause of the type 2N von Willebrand's disease in our cohort of patients. Using DMLE+ software and the mathematical model of Bengtsson and Thomson, it was estimated that the R854Q mutation occurred from 10,000 to 40,000 years ago, which is consistent with the short dimension of the haplotype shared by our patients. Together with the fact that the R854Q mutation seems to be limited to Caucasian populations, these findings suggest that a single mutational event took place after human populations moved from Africa towards Europe. -access paper. doi:10.3324/haematol.2012.066019 A common ancestor more than 10,000 years old for patients with R854Q-related type 2N von Willebrand's disease in Italy ©2013 Ferrata Storti Foundation. This is an open ABSTRACT Manuscript

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An Arg760Cys mutation in the consensus sequence of the von Willebrand factor propeptide cleavage site is responsible for a new von Willebrand disease variant

Cited by 43 publications

References 40 publications

A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

A von Willebrand factor fragment containing the D′D3 domains is sufficient to stabilize coagulation factor VIII in mice

Type 2N von Willebrand disease: Characterization and diagnostic difficulties

A common ancestor more than 10,000 years old for patients with R854Q-related type 2N von Willebrand's disease in Italy

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