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Thromb Haemost 2014; 112(01): 96-108
DOI: 10.1160/TH13-11-0902
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

von Willebrand disease type 2A phenotypes IIC, IID and IIE: A day in the life of shear-stressed mutant von Willebrand factor

Maria A. Brehm   *
1   Department of Paediatric Haematology and Oncology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
,
Volker Huck   *
4   Heidelberg University, Medical Faculty Mannheim, Experimental Dermatology, Mannheim, Germany
,
Camilo Aponte-Santamaría
3   Heidelberger Institut für Theoretische Studien, Heidelberg, Germany
,
Tobias Obser
1   Department of Paediatric Haematology and Oncology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
,
Sandra Grässle
4   Heidelberg University, Medical Faculty Mannheim, Experimental Dermatology, Mannheim, Germany
,
Florian Oyen
1   Department of Paediatric Haematology and Oncology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
,
Ulrich Budde
2   MEDILYS Laborgesellschaft mbH, Hemostaseology, Asklepios Klinik Altona, Hamburg, Germany
,
Sonja Schneppenheim
2   MEDILYS Laborgesellschaft mbH, Hemostaseology, Asklepios Klinik Altona, Hamburg, Germany
,
Carsten Baldauf
5   Fritz-Haber-Institut der Max-Planck-Gesellschaft, Berlin, Germany
,
Frauke Gräter
3   Heidelberger Institut für Theoretische Studien, Heidelberg, Germany
,
Stefan W. Schneider
4   Heidelberg University, Medical Faculty Mannheim, Experimental Dermatology, Mannheim, Germany
,
Reinhard Schneppenheim
1   Department of Paediatric Haematology and Oncology, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany
› Institutsangaben 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).
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Publikationsverlauf

Received: 04. November 2013

Accepted after major revision: 11. Februar 2014

Publikationsdatum:
01. Dezember 2017 (online)

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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.

Keywords

von Willebrand factor - VWD - shear flow - microfluidic - molecular dynamics simulations

* The first two authors contributed equally to this work.


 

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