Subscribe to RSS
DOI: 10.1160/TH13-11-0902
von Willebrand disease type 2A phenotypes IIC, IID and IIE: A day in the life of shear-stressed mutant von Willebrand factor
Financial support: This study was supported by research funding from the German Research Foundation (DFG) to the Research Group FOR1543: “Shear flow regulation of hemostasis – bridging the gap between nanomechanics and clinical presentation” (RS, MAB, TO, UB, VH, SWS, SG, CB, FG, CAS) and the SFB/Transregio23 (SWS TP A9).Publication History
Received:
04 November 2013
Accepted after major revision:
11 February 2014
Publication Date:
01 December 2017 (online)
Summary
The bleeding disorder von Willebrand disease (VWD) is caused by mutations of von Willebrand factor (VWF), a multimeric glycoprotein essential for platelet-dependent primary haemostasis. VWD type 2A–associated mutations each disrupt VWF biosynthesis and function at different stages, depending on the VWF domain altered by the mutation. These effects cause considerable heterogeneity in phenotypes and symptoms. To characterise the molecular mechanisms underlying the specific VWF deficiencies in VWD 2A/IIC, IID and IIE, we investigated VWF variants with patient-derived mutations either in the VWF pro-peptide or in domains D3 or CK. Additionally to static assays and molecular dynamics (MD) simulations we used microfluidic approaches to perform a detailed investigation of the shear-dependent function of VWD 2A mutants. For each group, we found distinct characteristics in their intracellular localisation visualising specific defects in biosynthesis which are correlated to respective multimer patterns. Using microfluidic assays we further determined shear flow-dependent characteristics in polymer-platelet-aggregate formation, platelet binding and string formation for all mutants. The phenotypes observed under flow conditions were not related to the mutated VWF domain. By MD simulations we further investigated how VWD 2A/IID mutations might alter the ability of VWF to form carboxy-terminal dimers. In conclusion, our study offers a comprehensive picture of shear-dependent and shear-independent dysfunction of VWD type 2A mutants. Furthermore, our microfluidic assay might open new possibilities for diagnosis of new VWD phenotypes and treatment choice for VWD patients with shear-dependent VWF dysfunctions that are currently not detectable by static tests.
* The first two authors contributed equally to this work.
-
References
- 1 Haberichter SL, Fahs SA, Montgomery RR. von Willebrand factor storage and multimerization: 2 independent intracellular processes. Blood 2000; 96: 1808-1815.
- 2 Wagner DD. Cell biology of von Willebrand factor. Annu Rev Cell Biol 1990; 06: 217-246.
- 3 Metcalf DJ, Nightingale TD, Zenner HL. et al. Formation and function of Weibel-Palade bodies. J Cell Sci 2008; 121: 19-27.
- 4 Alexander-Katz A, Schneider MF, Schneider SW. et al. Shear-flow-induced unfolding of polymeric globules. Phys Rev Lett 2006; 97: 138101.
- 5 Schneider SW, Nuschele S, Wixforth A. et al. Shear-induced unfolding triggers adhesion of von Willebrand factor fibers. Proc Natl Acad Sci USA 2007; 104: 7899-7903.
- 6 Singh I, Themistou E, Porcar L. et al. Fluid shear induces conformation change in human blood protein von Willebrand factor in solution. Biophys J 2009; 96: 2313-2320.
- 7 Cheng H, Yan R, Li S. et al. Shear-induced interaction of platelets with von Willebrand factor results in glycoprotein Ibalpha shedding. Am J Physiol Heart Circ Physiol 2009; 297: H2128-2135.
- 8 Ruggeri ZM. Structure of von Willebrand factor and its function in platelet adhesion and thrombus formation. Best Pract Res Clin Haematol 2001; 14: 257-279.
- 9 Schneppenheim R, Budde U. von Willebrand factor: the complex molecular genetics of a multidomain and multifunctional protein. J Thromb Haemost 2011; 09 (Suppl. 01) 209-215.
- 10 Brehm MA, Schenk TMH, Zhou X. et al. Intracellular localization of human Ins(1,3,4,5,6)P-5 2-kinase. Biochem J 2007; 408: 335-345.
- 11 Budde U, Schneppenheim R, Eikenboom J. et al. 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 Thromb Haemost 2008; 06: 762-771.
- 12 Budde U, Schneppenheim R, Plendl H. et al. Luminographic detection of von Willebrand factor multimers in agarose gels and on nitrocellulose membranes. Thromb Haemost 1990; 63: 312-315.
- 13 Schneppenheim R, Plendl H, Budde U. Luminography-an alternative assay for detection of von Willebrand factor multimers. Thromb Haemost 1988; 60: 133-136.
- 14 Daopin S, Piez KA, Ogawa Y. et al. Crystal structure of transforming growth factor-beta 2: an unusual fold for the superfamily. Science 1992; 257: 369-373.
- 15 Meitinger T, Meindl A, Bork P. et al. Molecular modelling of the Norrie disease protein predicts a cystine knot growth factor tertiary structure. Nat Genet 1993; 05: 376-380.
- 16 Zhou YF, Eng ET, Zhu J. et al. Sequence and structure relationships within von Willebrand factor. Blood 2012; 120: 449-458.
- 17 Katsumi A, Tuley EA, Bodo I. et al. Localization of disulfide bonds in the cystine knot domain of human von Willebrand factor. J Biol Chem 2000; 275: 25585-25594.
- 18 Hess B, Kutzner C, van der Spoel D. et al. GROMACS 4: Algorithms for highly efficient, load-balanced, and scalable molecular simulation. J Chem Theory Comput 2008; 04: 435-447.
- 19 Van der Spoel D, Lindahl E, Hess B. et al. GROMACS: Fast, flexible, and free. J Comput Chem 2005; 26: 1701-1718.
- 20 Krivobokova T, Briones R, Hub JS. et al. Partial least-squares functional mode analysis: application to the membrane proteins AQP1, Aqy1, and CLC-ec1. Biophys J 2012; 103: 786-796.
- 21 Rickham PP. Human Experimentation. Code of Ethics of the World Medical Association. Declaration of Helsinki. Br Med J 1964; 02: 177.
- 22 De Ceunynck K, Rocha S, Feys HB. et al. Local elongation of endothelial cell-anchored von Willebrand factor strings precedes ADAMTS13 protein-mediated proteolysis. J Biol Chem 2011; 286: 36361-36367.
- 23 Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012; 09: 671-675.
- 24 dos Santos SM, Klinkhardt U, Schneppenheim R. et al. Using ImageJ for the quantitative analysis of flow-based adhesion assays in real-time under physiologic flow conditions. Platelets 2010; 21: 60-66.
- 25 Schneppenheim R, Brassard J, Krey S. et al. Defective dimerization of von Willebrand factor subunits due to a Cys-> Arg mutation in type IID von Willebrand disease. Proc Natl Acad Sci USA 1996; 93: 3581-3586.
- 26 Wang JW, Groeneveld DJ, Cosemans G. et al. Biogenesis of Weibel-Palade bodies in von Willebrand’s disease variants with impaired von Willebrand factor intrachain or interchain disulfide bond formation. Haematologica 2012; 97: 859-866.
- 27 Zhou YF, Springer TA. Highly reinforced structure of a C-terminal dimerization domain in von Willebrand factor. Blood. 2014 Epub ahead of print
- 28 Allen S, Abuzenadah AM, Hinks J. et al. A novel von Willebrand disease-causing mutation (Arg273Trp) in the von Willebrand factor propeptide that results in defective multimerization and secretion. Blood 2000; 96: 560-568.
- 29 Gaucher C, Dieval J, Mazurier C. Characterization of von Willebrand factor gene defects in two unrelated patients with type IIC von Willebrand disease. Blood 1994; 84: 1024-1030.
- 30 Schneppenheim R, Thomas KB, Krey S. et al. Identification of a candidate missense mutation in a family with von Willebrand disease type IIC. Hum Genet 1995; 95: 681-686.
- 31 Schneppenheim R, Michiels JJ, Obser T. et al. A cluster of mutations in the D3 domain of von Willebrand factor correlates with a distinct subgroup of von Willebrand disease: type 2A/IIE. Blood 2010; 115: 4894-4901.
- 32 Chen H, Fallah MA, Huck V. et al. Blood-clotting-inspired reversible polymercolloid composite assembly in flow. Nat Commun 2013; 04: 1333.
- 33 Schneppenheim R, Budde U, Obser T. et al. Expression and characterization of von Willebrand factor dimerization defects in different types of von Willebrand disease. Blood 2001; 97: 2059-2066.
- 34 Enayat MS, Guilliatt AM, Surdhar GK. et al. Aberrant dimerization of von Willebrand factor as the result of mutations in the carboxy-terminal region: identification of 3 mutations in members of 3 different families with type 2A (phenotype IID) von Willebrand disease. Blood 2001; 98: 674-680.
- 35 Tjernberg P, Vos HL, Spaargaren-van Riel CC. et al. Differential effects of the loss of intrachain- versus interchain-disulfide bonds in the cystine-knot domain of von Willebrand factor on the clinical phenotype of von Willebrand disease. Thromb Haemost 2006; 96: 717-724.
- 36 Haberichter SL, Budde U, Obser T. et al. The mutation N528S in the von Willebrand factor (VWF) propeptide causes defective multimerization and storage of VWF. Blood 2010; 115: 4580-4587.