Molecular characterization, recombinant protein expression, and mRNA analysis of type 3 von Willebrand disease: Studies of an Italian cohort of 10 patients.

Abstract

Type 3 von Willebrand disease (VWD3) is characterized by unmeasurable von Willebrand factor (VWF) levels in plasma and platelets and severe but variable hemorrhagic symptoms. To identify and characterize the causal mutations, we screened 10 Italian patients with VWD3 by several techniques including Multiplex Ligation-dependent Probe Amplification to identify large insertions and deletions, High Resolution Melting and PCR coupled with Sanger sequencing. Fourteen different mutations scattered throughout the VWF gene were identified, 10 of which were novel. As expected, most of these mutations caused null alleles: five were deletions (del exons 1-3, del exon 17, c.2157delA, c.2269delCT, and c.3940delG), three nonsense (p.Q1526X, p.E1549X, and p.C2448X) and three potential splice-site mutations (c.658-2A>G, c.7729+7C>T, and c.8155+1G>T). Three candidate missense mutations (p.C2184S, p.C2212R, and p.C2325S) were also identified. Missense mutations and the putative splice-site defects were confirmed to be disease related by in vitro expression studies and mRNA analysis. None of these patients have developed alloantibodies against VWF. This study extends our previous finding that most of the mutations that we identified in VWD3 patients arise independently and are scattered throughout the entire VWF gene.

• Authors: Maria Solimando, Luciano Baronciani, Silvia La Marca, Giovanna Cozzi, Rosanna Asselta, Maria Teresa Canciani, Augusto B Federici, Flora Peyvandi
• Journal: American journal of hematology (Am J Hematol) Vol. 87 Issue 9 Pg. 870-4 (Sep 2012) ISSN: 1096-8652 [Electronic] United States
• PMID: 22674667 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 Wiley Periodicals, Inc.

Chemical References

• DNA, Complementary
• RNA Splice Sites
• RNA, Messenger
• von Willebrand Factor

Topics

• Adolescent
• Adult
• Animals
• COS Cells
• Cell Culture Techniques
• Child
• Chlorocebus aethiops
• Cohort Studies
• DNA, Complementary
• Female
• Humans
• Italy
• Male
• Mutation
• Mutation, Missense
• Plasmids
• RNA Splice Sites
• RNA, Messenger (genetics)
• Transfection
• Young Adult
• von Willebrand Disease, Type 3 (blood, genetics)
• von Willebrand Factor (biosynthesis, genetics)