A novel mouse model of type 2N VWD was developed by CRISPR/Cas9 gene editing and recapitulates human type 2N VWD.

Autor:	Shi Q; Versiti Blood Research Institute, Milwaukee, WI.; Departments of Pediatrics, Physiology, and Genetics, Medical College of Wisconsin, Milwaukee, WI.; Children's Research Institute, Milwaukee, WI; and.; MACC Fund Research Center, Milwaukee, WI., Fahs SA; Versiti Blood Research Institute, Milwaukee, WI., Mattson JG; Versiti Blood Research Institute, Milwaukee, WI., Yu H; Versiti Blood Research Institute, Milwaukee, WI.; Departments of Pediatrics, Physiology, and Genetics, Medical College of Wisconsin, Milwaukee, WI., Perry CL; Versiti Blood Research Institute, Milwaukee, WI., Morateck PA; Versiti Blood Research Institute, Milwaukee, WI., Schroeder JA; Versiti Blood Research Institute, Milwaukee, WI.; Departments of Pediatrics, Physiology, and Genetics, Medical College of Wisconsin, Milwaukee, WI., Rapten J; Versiti Blood Research Institute, Milwaukee, WI., Weiler H; Versiti Blood Research Institute, Milwaukee, WI.; Departments of Pediatrics, Physiology, and Genetics, Medical College of Wisconsin, Milwaukee, WI., Montgomery RR; Versiti Blood Research Institute, Milwaukee, WI.; Departments of Pediatrics, Physiology, and Genetics, Medical College of Wisconsin, Milwaukee, WI.; Children's Research Institute, Milwaukee, WI; and.; MACC Fund Research Center, Milwaukee, WI.
Jazyk:	angličtina
Zdroj:	Blood advances [Blood Adv] 2022 May 10; Vol. 6 (9), pp. 2778-2790.
DOI:	10.1182/bloodadvances.2021006353
Abstrakt:	Type 2N von Willebrand disease is caused by mutations in the factor VIII (FVIII) binding site of von Willebrand factor (VWF), resulting in dysfunctional VWF with defective binding capacity for FVIII. We developed a novel type 2N mouse model using CRISPR/Cas9 technology. In homozygous VWF2N/2N mice, plasma VWF levels were normal (1167 ± 257 mU/mL), but the VWF was completely incapable of binding FVIII, resulting in 53 ± 23 mU/mL of plasma FVIII levels that were similar to those in VWF-deficient (VWF-/-) mice. When wild-type human or mouse VWF was infused into VWF2N/2N mice, endogenous plasma FVIII was restored, peaking at 4 to 6 hours post-infusion, demonstrating that FVIII expressed in VWF2N mice is viable but short-lived unprotected in plasma due to dysfunctional 2N VWF. The whole blood clotting time and thrombin generation were impaired in VWF2N/2N but not in VWF-/- mice. Bleeding time and blood loss in VWF2N/2N mice were similar to wild-type mice in the lateral tail vein or ventral artery injury model. However, VWF2N/2N mice, but not VWF-/- mice, lost a significant amount of blood during the primary bleeding phase after a tail tip amputation injury model, indicating that alternative pathways can at least partially restore hemostasis when VWF is absent. In summary, we have developed a novel mouse model by gene editing with both the pathophysiology and clinical phenotype found in severe type 2N patients. This unique model can be used to investigate the biological properties of VWF/FVIII association in hemostasis and beyond. (© 2022 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
Databáze:	MEDLINE
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