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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2008;28:419.)
© 2008 American Heart Association, Inc.

Integrative Physiology/Experimental Medicine

Correction of Bleeding Symptoms in von Willebrand Factor–Deficient Mice by Liver-Expressed von Willebrand Factor Mutants

Isabelle Marx; Peter J. Lenting; Thure Adler; Ronan Pendu; Olivier D. Christophe; Cécile V. Denis

From the Institut National de la Santé et de la Recherche Médicale (INSERM) U770 (I.M., T.A., O.D.C., C.V.D.), Le Kremlin-Bicêtre, F-94276 France; Univ Paris-Sud, Le Kremlin-Bicêtre, F-94276 France; and the Laboratory for Thrombosis and Haemostasis (P.J.L., R.P.), Department of Clinical Chemistry & Haematology, University Medical Center Utrecht, The Netherlands.

Correspondence to Cécile V. Denis, INSERM U770, 80 rue du Général Leclerc, 94276 Le Kremlin-Bicêtre Cedex, France. E-mail denis{at}kb.inserm.fr

Abstract

Objective— von Willebrand Factor (vWF) structure-function relationship has been studied only in vitro. To investigate the physiological importance of particular vWF domains, we have introduced mutations into murine vWF (mvWF) cDNA inhibiting vWF binding to glycoprotein (Gp) Ib, GpIIbIIIa, and to fibrillar collagen.

Methods and Results— We delivered wild-type (WT) or mutant mvWF cDNA into vWF-deficient (Vwf^–/–) mice using hydrodynamic injection and assessed whether hemorrhagic symptoms could be corrected. Hydrodynamic gene transfer resulted in high expression of plasma mvWF 24 hours after injection (438±63% for 50 µg of cDNA). Factor VIII activity was normalized in Vwf^–/– mice injected with mvWF cDNA and multimerization was achieved. Bleeding time was corrected after injection of WT mvWF cDNA in Vwf^–/– mice whereas noninjected mice did not stop bleeding. Injection of the GpIIbIIIa and the collagen binding mutants in Vwf^–/– mice also resulted in a correction of bleeding time whereas mice injected with the GpIb binding mutant were bleeding for as long they were observed, although blood loss was decreased compared with noninjected mice (61±21 µL versus 232±63 µL).

Conclusion— Our model allows the rapid in vivo evaluation of specific mutations on plasma vWF function.

To study von Willebrand factor (vWF) structure-function relationship in vivo, we have expressed wild-type and mutant vWF by hydrodynamic injection in vWF-deficient mice. Wild-type vWF and vWF carrying mutations in its collagen and platelet glycoprotein (GP) IIbIIIa binding sites were able to correct bleeding symptoms in the knockout mice.

Key Words: von Willebrand disease • animal model • vWF • bleeding • hydrodynamic injection

This article has been cited by other articles:

				I. Marx, O. D. Christophe, P. J. Lenting, A. Rupin, M.-O. Vallez, T. J. Verbeuren, and C. V. Denis Altered thrombus formation in von Willebrand factor-deficient mice expressing von Willebrand factor variants with defective binding to collagen or GPIIbIIIa Blood, August 1, 2008; 112(3): 603 - 609. [Abstract] [Full Text] [PDF]