Novel Large Deletion Is the Most Frequent Cause of Type 3 von Willebrand Disease in Hungary.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1013-1013
Author(s):  
Adrien Mohl ◽  
Tamás Masszi ◽  
Eszter Nagy ◽  
Tobias Obser ◽  
Florian Oyen ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Large Deletion ◽  
Severe Form ◽  
Von Willebrand Disease ◽  
Rational Approach ◽  
Molecular Testing ◽  
Spontaneous Bleeding ◽  
Virtual Absence ◽  
Type 3 ◽  
Von Willebrand

Abstract Background: Type 3 is the most severe form of von Willebrand disease (VWD) caused by the virtual absence of von Willebrand factor (VWF) in affected patients. The prevalence of type 3 VWD in Hungary is 2.6 per million. Capitalizing on a nationwide National Bleeding Disorder Registry, we designed a study to characterize the genetic background of the entire Hungarian type 3 VWD population. The current report focuses on the molecular characterization of a novel large deletion. Methods: 24 patients from 23 unrelated families were studied by direct sequencing of the 52 exons of the VWF gene. The breakpoints of a large deletion were characterized by standard gene mapping. Breakpoint-specific PCR was used to confirm the presence of the deletion, and to screen for identical deletions in other populations from Germany, Russia, and Poland. Results: A large partial deletion (delExon1-3) of the 5′-region of the VWF gene was detected in 10 alleles (19 percent of all type 3 mutations). Five patients from 4 unrelated families were homozygous, and 2 patients were heterozygous for the deletion. Consanguinity was known in one of the families. In comparison, 2435 delC in exon 18, a common cause of type 3 VWD in some European populations, was found on 6 alleles (12 percent; one patient homozygous). The large deletion resulted in the loss of a 35 kb fragment, incorporating exons 1, 2 and 3. The 5′ breakpoint is located in the 5′ untranslated region, while the 3′ breakpoint is in intron 3 of VWF. No other known gene is lost with the deletion. Clinically, all homozygous patients had serious bleeding episodes from infancy requiring frequent VWF substitutions. However, bleeding became much milder in all patients with no significant spontaneous bleeding after the age of 3-5 years. No inhibitor to VWF was detected. delExon1-3 was not detected in any of the other screened populations. Conclusion: We report a large novel deletion including exons 1, 2 and 3 of VWF commonly causing type 3 VWD in the Hungarian population. This mutation, which is most probably due to a founder effect, seems to be unique to Hungarian patients with a high allele frequency. Together, delExon1-3 and 2435delC make up 31 % of genetic defects in Hungarian patients with VWD type 3. This offers a rational approach to molecular testing of respective families in Hungary.

The Use of High Resolution Melting (HRM) Analysis for Molecular Gene Defects of Type 3 Von Willebrand Disease: Studies of An Italian Cohort of 10 Patients.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3498-3498
Author(s):  
Maria Solimando ◽  
Augusto B. Federici ◽  
Luciano Baronciani ◽  
Alberto Mussetti ◽  
Margherita Punzo ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Large Deletion ◽  
Von Willebrand Disease ◽  
Missense Mutations ◽  
Duplex Pcr ◽  
Hrm Analysis ◽  
Large Deletions ◽  
Large Gene ◽  
Type 3 ◽  
Von Willebrand

Abstract Abstract 3498 Poster Board III-435 Introduction Type 3 von Willebrand disease (VWD3) is a severe autosomal recessive inherited bleeding disorder caused by a virtually complete absence of von Willebrand Factor (VWF). Classically, patients are homozygous or compound heterozygous for null alleles due to nonsense mutations, small insertions/deletions, splice site defects or, more rarely, large gene deletions spread throughout the VWF gene. Nevertheless, several missense mutations have also been reported. Aims of the study, patients and methods The aim of this study was to investigate the molecular basis of VWD3 in 10 Italian patients using DNA direct sequencing, High Resolution Melting (HRM) analysis and duplex PCR. HRM is a simple, low-cost, and rapid PCR-based method for detecting sequence variation by measuring changes in the melting temperature of double stranded DNA. Duplex PCR was used to screen for the presence of some known large deletions causing VWD3: the 61-kb deletion encompassing exons 6-16 (Xie et al. Blood Cells Mol Dis. 2006; 36: 385), the 253-kb deletion involving the whole VWF gene (Schneppenheim et al. J Thromb Haemost. 2007; 5: 722), the exons 1-3 deletion (Mohl et al. J Thromb Haemost. 2008; 6: 1729), and the exons 4-5 deletion (Sutherland et al. Blood. 2009; 114: 1091). Results and discussion Twenty-four exons were analyzed by direct sequencing, 21 exons by HRM and, so far, 6 exons using both methods. The following mutations were identified in 8 of the 10 patients investigated: 2157delA/7729+7C>T; C2184S*/undetermined; Q1526X*/C2325S*; del ex1-3/3940delG*; 8155+1G>T*/8155+1G>T*; E1549X*/undetermined; 658-2A>G*/658-2A>G*; del ex 1-3/undetermined. Direct sequencing revealed 7 mutations, HRM analysis could detect 2 defects (2157delA, C2325S) and duplex PCR identified one large deletion. Seven of these 11 mutations were novel (indicated with *). Two patients were found to carry mutations in the homozygous state. To confirm these findings, their parents will have to be investigated in order to exclude the presence of a large gene deletion in one of the alleles. Interestingly, the large deletion involving exons 1-3, which was previously reported in the Hungarian population, was also found in 2 unrelated patients. Two missense mutations were identified, both involving a cysteine residue, further suggesting the importance of these residues in the correct folding/processing/secretion of the neo-synthesized VWF. In those patients who still remain uncharacterized further analysis should be performed to search for intronic mutations or heterozygous large deletions responsible for aberrant splicing/post-transcriptional events. Conclusion Based on these preliminary data, HRM analysis, to our knowledge used for the first time in the molecular diagnosis of VWD3, in our hands seems to be an accurate and rapid method for mutational screening of VWF gene. However, so far, the presence of many polymorphic sites in the VWF coding region has strongly limited the use of this technique to 21 exons of the gene. Disclosures: Baronciani: Bayer Awards: Research Funding.

scholarly journals Rare Occurrence of Inhibitors in Von Willebrand Disease: A Case Report

2022 ◽  
Vol 8 ◽  
Author(s):  
Bipin P. Kulkarni ◽  
Kirti Ghargi ◽  
Chandrakala Shanmukhaiah ◽  
Shrimati D. Shetty
Keyword(s):  
Correct Diagnosis ◽  
Severe Form ◽  
Von Willebrand Disease ◽  
Rare Occurrence ◽  
Standard Assay ◽  
Type 3 ◽  
Von Willebrand ◽  
High Degree ◽  
Willebrand Factor ◽  
Inhibitor Titer

Introduction: Type 3 Von Willebrand Disease (VWD) is the least common but the most severe form of a disease, with a prevalence of about 0. 5 to 1 per million in Western countries. The prevalence of type 3 VWD in the developing countries, with a high degree of consanguinity, is about 6 per million. Moreover, due to underdiagnosis of the milder cases, the prevalence of type 3 VWD is about 50% of the cases. Rarely, some patients develop the Von Willebrand Factor (VWF) inhibitors, which may subsequently develop severe anaphylactic reactions on further exposure to the VWF containing factor replacement therapy. The prevalence of inhibitor development in patients with type 3 VWD has been shown to be in the range of 5.8 to 9.5%. In the absence of a gold standard assay for the quantitation of VWF inhibitors, a correct diagnosis and management of these patients are often challenging.Objectives: The objective of this study is to standardize the Bethesda assay for the VWF inhibitors and to estimate the VWD inhibitor titer in two cases of congenital type 3 VWD, which developed the VWF inhibitors.Results and Conclusions: We could successfully standardize the Bethesda assay for the quantitation of VWF inhibitors in two patients with congenital type 3 VWD with inhibitors.

An Alu-mediated novel large deletion is the most frequent cause of type 3 von Willebrand disease in Hungary

2008 ◽  
Vol 6 (10) ◽  
pp. 1729-1735 ◽  
Author(s):  
A. MOHL ◽  
R. MARSCHALEK ◽  
T. MASSZI ◽  
E. NAGY ◽  
T. OBSER ◽  
...  
Keyword(s):  
Large Deletion ◽  
Von Willebrand Disease ◽  
Type 3 ◽  
Von Willebrand

P061 Type 3 von Willebrand disease in Hungary: A partial large deletion is the most common genetic defect

Blood Reviews ◽  
2007 ◽  
Vol 21 ◽  
pp. S105
Author(s):  
A. Mohl ◽  
Z. Boda ◽  
R. Jager ◽  
H. Losonczy ◽  
A. Marosi ◽  
...  
Keyword(s):  
Genetic Defect ◽  
Large Deletion ◽  
Von Willebrand Disease ◽  
Type 3 ◽  
Von Willebrand

Update on Molecular Testing in von Willebrand Disease

2019 ◽  
Vol 45 (07) ◽  
pp. 708-719 ◽  
Author(s):  
Javier Batlle ◽  
Almudena Pérez-Rodríguez ◽  
Irene Corrales ◽  
Nina Borràs ◽  
Joana Costa Pinto ◽  
...  
Keyword(s):  
Hemophilia A ◽  
Von Willebrand Disease ◽  
Molecular Testing ◽  
History Of ◽  
Pros And Cons ◽  
Phenotypic Tests ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

AbstractDiagnosis of von Willebrand disease (VWD) depends on personal and family history of bleeding and confirmatory laboratory testing. Currently available phenotypic tests for VWD contain potential sources for error that may distort results. Despite an exponential growth of information about the von Willebrand factor gene (VWF), the role of molecular diagnosis in VWD is still controversial. Due to the complexity and high cost of conventional molecular analyses, some investigators have recommended limiting this approach to distinguish suspected type 2N VWD from hemophilia A, type 2B from platelet-type VWD, and the exploration of type 3 VWD. New genetic methodologies and approaches are becoming available, but there is still some reluctance for their implementation in VWD diagnosis. This article discusses the pros and cons of molecular testing in VWD considering the experience obtained through the multicenter project “Molecular and Clinical Profile of VWD in Spain (PCM-EVW-ES).”

Phenotypic and molecular characterisation of type 3 von Willebrand disease in a cohort of Indian patients

2013 ◽  
Vol 109 (04) ◽  
pp. 652-660 ◽  
Author(s):  
Ulrich Budde ◽  
Rifat Jan ◽  
Florian Oyen ◽  
Meganathan Kannan ◽  
Reinhard Schneppenheim ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Mutation Screening ◽  
Strand Break ◽  
Von Willebrand Disease ◽  
Missense Mutations ◽  
Large Deletions ◽  
High Propensity ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

SummarySevere type 3 VWD (VWD3) is characterised by complete absence or presence of trace amounts of non-functional von Willebrand factor (VWF). The study was designed to evaluate the VWF mutations in VWD3 patients and characterise the breakpoints of two identified homozygous novel large deletions. Patients were diagnosed by conventional tests and VWF multimer analysis. Mutation screening was performed in 19 VWD3 patients by direct sequencing of VWF including flanking intronic sequence and multiplex ligation-dependent probe amplification (MLPA) analysis. Breakpoint characterisation of two identified novel large deletions was done using walking primers and long spanning PCR. A total of 21 different mutations including 15 (71.4%) novel ones were identified in 17 (89.5%) patients. Of these mutations, five (23.8%) were nonsense (p.R1659*, p.R1779*, p.R1853*, p.Q2470*, p.Q2520*), one was a putative splice site (p.M814I) and seven (33.3%) were deletions (p.L254fs*48, p.C849fs*60, p.L1871fs*6, p.E2720fs*24) including three novel large deletions of exon 14–15, 80,830bp (−41510_657+7928A*del) and 2,231bp [1534–2072T_c.1692G*del(p.512fs*terminus)] respectively. A patient carried gene conversion comprising of pseudogene harbouring mutations. The missense mutations (p.G19R, p.K355R, p.D437Y, p.C633R, p.M771V, p.G2044D, p.C2491R) appear to play a major role and were identified in seven (36.8%) patients. In conclusion, a high frequency of novel mutations suggests the high propensity of VWF for new mutations. Missense and deletion mutations found to be a common cause of VWD3 in cohort of Indian VWD3 patients. Breakpoints characterisation of two large deletions reveals the double strand break and non-homologous recombination as deletions mechanism.

Treatment of Von Willebrand Disease

2001 ◽  
Vol 86 (07) ◽  
pp. 149-153 ◽  
Author(s):  
Pier Mannuccio Mannucci
Keyword(s):  
Factor Viii ◽  
Von Willebrand Factor ◽  
Von Willebrand Disease ◽  
Spontaneous Bleeding ◽  
Primary Hemostasis ◽  
Bleeding Prophylaxis ◽  
Bleeding Episodes ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryIn von Willebrand disease, there are two main options for the treatment of spontaneous bleeding episodes and for bleeding prophylaxis: desmopressin and transfusional therapy with plasma products. Desmopressin is the treatment of choice for most patients with type 1, who account for approximately 70 to 80 per cent of all cases with the disease. This non-transfusional hemostatic agent raises endogenous factor VIII and von Willebrand factor three- to fivefold and thereby transiently corrects both the intrinsic coagulation and primary hemostasis defects. In patients with the more severe type 3 and in the majority of those with type 2 desmopressin is not effective or is contraindicated, so that it is usually necessary to resort to plasma concentrates containing factor VIII and von Willebrand factor. Concentrates treated with virus inactivation methods should be preferred to cryoprecipitate because they are equally effective and perceived as safer.

A novel deletion mutation is recurrent in von Willebrand disease types 1 and 3

Blood ◽  
2009 ◽  
Vol 114 (5) ◽  
pp. 1091-1098 ◽  
Author(s):  
Megan S. Sutherland ◽  
Anthony M. Cumming ◽  
Mackenzie Bowman ◽  
Paula H. B. Bolton-Maggs ◽  
Derrick J. Bowen ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Direct Sequencing ◽  
Von Willebrand Disease ◽  
Compound Heterozygous ◽  
Asian Origin ◽  
Expression Studies ◽  
Autosomal Dominant Fashion ◽  
Type 3 ◽  
Von Willebrand

Direct sequencing of VWF genomic DNA in 21 patients with type 3 von Willebrand disease (VWD) failed to reveal a causative homozygous or compound heterozygous VWF genotype in 5 cases. Subsequent analysis of VWF mRNA led to the discovery of a deletion (c.221-977_532 + 7059del [p.Asp75_Gly178del]) of VWF in 7 of 12 white type 3 VWD patients from 6 unrelated families. This deletion of VWF exons 4 and 5 was absent in 9 patients of Asian origin. We developed a genomic DNA-based assay for the deletion, which also revealed its presence in 2 of 34 type 1 VWD families, segregating with VWD in an autosomal dominant fashion. The deletion was associated with a specific VWF haplotype, indicating a possible founder origin. Expression studies indicated markedly decreased secretion and defective multimerization of the mutant VWF protein. Further studies have found the mutation in additional type 1 VWD patients and in a family expressing both type 3 and type 1 VWD. The c.221-977_532 + 7059del mutation represents a previously unreported cause of both types 1 and 3 VWD. Screening for this mutation in other type 1 and type 3 VWD patient populations is required to elucidate further its overall contribution to VWD arising from quantitative deficiencies of VWF.

scholarly journals Successful Aortic Aneurysm Repair in a Woman with Severe von Willebrand (Type 3) Disease

2015 ◽  
Vol 2015 ◽  
pp. 1-8
Author(s):  
Victoria Campbell ◽  
Kevin Marriott ◽  
Rex Stanbridge ◽  
Abdul Shlebak
Keyword(s):  
Aortic Aneurysm ◽  
Severe Form ◽  
Von Willebrand Disease ◽  
Severe Bleeding ◽  
Recessive Trait ◽  
Aneurysm Repair ◽  
Factor Concentrate ◽  
Type 3 ◽  
Von Willebrand ◽  
Aortic Aneurysm Repair

von Willebrand disease type 3 (VWD3) is a rare but the most severe form of von Willebrand disease; it is due to almost complete lack of von Willebrand factor activity (VWF:RCo). It is inherited as autosomal recessive trait; whilst heterozygote carriers have mild, or no symptoms, patients with VWD3 show severe bleeding symptoms. In the laboratory, this is characterised by undetectable VWF:Ag, VWF:RCo, and reduced levels of factor VIII < 0.02 IU/dL. The bleeding is managed with von Willebrand/FVIII factor concentrate replacement therapy. In this rare but challenging case we report on the successful excision and repair of an ascending aortic aneurysm following adequate VWF/FVIII factor concentrate replacement using Haemate-P.

How I treat patients with von Willebrand disease

Blood ◽  
2001 ◽  
Vol 97 (7) ◽  
pp. 1915-1919 ◽  
Author(s):  
Pier Mannuccio Mannucci
Keyword(s):  
Factor Viii ◽  
Von Willebrand Factor ◽  
Von Willebrand Disease ◽  
Spontaneous Bleeding ◽  
Primary Hemostasis ◽  
Bleeding Prophylaxis ◽  
Bleeding Episodes ◽  
Type 3 ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract Von Willebrand disease (vWD) is a frequent inherited disorder of hemostasis that affects both sexes. Two abnormalities are characteristic of the disease, which is caused by a deficiency or a defect in the multimeric glycoprotein called von Willebrand factor: low platelet adhesion to injured blood vessels and defective intrinsic coagulation owing to low plasma levels of factor VIII. There are 2 main options available for the treatment of spontaneous bleeding episodes and for bleeding prophylaxis: desmopressin and transfusional therapy with plasma products. Desmopressin is the treatment of choice for most patients with type 1 vWD, who account for approximately 70% to 80% of cases. This nontransfusional hemostatic agent raises endogenous factor VIII and von Willebrand factor 3 to 5 times and thereby corrects both the intrinsic coagulation and the primary hemostasis defects. In patients with the more severe type 3 and in most patients with type 2 disease, desmopressin is ineffective or is contraindicated and it is usually necessary to resort to plasma concentrates containing both factor VIII and von Willebrand factor. Concentrates treated with virucidal methods should be preferred to cryoprecipitate because they are equally effective and are perceived as safer.

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