Von Willebrand disease (VWD) is the most prevalent bleeding disorder in the world, affecting approximately 1% of the human population. It is divided into three general categories, types 1, 2 and 3. In type 2 VWD, most mutations result in replacement of a single amino acid in VWF. The functional consequences of these mutations are difficult to predict because of the complex nature of VWF synthesis and structure. The problem of type 2 VWD will be approached in this application with two specific aims.
In Specific Aim 1, a variety of approaches are proposed to examine the effects of type 2B gain-of-function mutations on the disulfide bond structure of the VWF A1-A2-A3 region. These studies address the hypothesis that mutations that change the number of Cys residues or increase the negative charge in the region of the Cys1272-Cys1458 disulfide bond alter the disulfide-bond structure of the region. The disulfide-bond structure of 2B VWF mutants will be examined by mass spectrometry (MS) and correlated with functional alterations.
In Specific Aim 2, plasma from a large number of well-characterized patients from a type 2 VWD kindred will be evaluated with a mathematical index of VWF quality and quantity that takes into account the percentage of mutant monomers incorporated into multimers, the relative content of multimers of different sizes, and the VWF quantity. This index will be correlated with VWF functional parameters and with the bleeding phenotype as quantified with a validated bleeding score questionnaire. These studies will be complemented with a longitudinal study of six patients with type 2B VWD (mutation R1308C) to compare the index with contemporaneously determined parameters of VWF function. It is expected that these studies will delineate both intra- and inter-individual variation in the VWF quality index. In aggregate, the proposed studies will shed light on the complex pathophysiology of type 2 VWD and have the potential to provide an accurate and quantitative assessment of bleeding risk in VWD patients.

Public Health Relevance

This application will examine the ways in which mutations alter the function of von Willebrand factor (VWF) and cause of the most common inherited bleeding disorder of humans, von Willebrand disease. This work will define how the mutations change VWF function and is expected to produce tests that can be used to predict a patient's risk of bleeding, which cannot be determined simply by knowing which mutation causes the disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL117639-05
Application #
9223729
Study Section
Special Emphasis Panel (ZRG1-VH-J (02)M)
Program Officer
Kindzelski, Andrei L
Project Start
2013-02-01
Project End
2018-01-31
Budget Start
2017-02-01
Budget End
2018-01-31
Support Year
5
Fiscal Year
2017
Total Cost
$579,456
Indirect Cost
$217,766
Name
Puget Sound Blood Center
Department
Type
Research Institutes
DUNS #
092881085
City
Seattle
State
WA
Country
United States
Zip Code
98104
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