title: A genetically-engineered von Willebrand disease type 2B mouse model displays defects in hemostasis and inflammation.
abstract: von Willebrand disease (VWD)-type 2B is characterized by gain-of-function mutations in the von Willebrand factor (VWF) A1-domain, leading to increased affinity for its platelet-receptor, glycoprotein Ibα. We engineered the first knock-in (KI) murine model for VWD-type 2B by introducing the p.V1316M mutation in murine VWF. Homozygous KI-mice replicated human VWD-type 2B with macrothrombocytopenia (platelet counts reduced by 55%, platelet volume increased by 44%), circulating platelet-aggregates and a severe bleeding tendency. Also, vessel occlusion was deficient in the FeCl3-induced thrombosis model. Platelet aggregation induced by thrombin or collagen was defective for KI-mice at all doses. KI-mice manifested a loss of high molecular weight multimers and increased multimer degradation. In a model of VWF-string formation, the number of platelets/string and string-lifetime were surprisingly enhanced in KI-mice, suggesting that proteolysis of VWF/p.V1316M is differentially regulated in the circulation versus the endothelial surface. Furthermore, we observed increased leukocyte recruitment during an inflammatory response induced by the reverse passive Arthus reaction. This points to an active role of VWF/p.V1316M in the exfiltration of leukocytes under inflammatory conditions. In conclusion, our genetically-engineered VWD-type 2B mice represent an original model to study the consequences of spontaneous VWF-platelet interactions and the physiopathology of this human disease.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Adam
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Caterina
authors.initials: C
authors.lastname: Casari
authors.affiliation: Kyoto Gakuen University, Kameoka, Japan.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Prévost
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Kauskot
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Loubière
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Paulette
authors.initials: P
authors.lastname: Legendre
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Repérant
authors.affiliation: INSERM UMR_S1140, 75006 Paris, France.|University Paris-Descartes, 75006 Paris, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Baruch
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Rosa
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Marijke
authors.initials: M
authors.lastname: Bryckaert
authors.affiliation: Department of Clinical Chemistry and Haematology, University of Utrecht, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Philip G
authors.initials: PG
authors.lastname: de Groot
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Olivier D
authors.initials: OD
authors.lastname: Christophe
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Lenting
authors.affiliation: Institut National de la Santé et de la Recherche Médicale, UMR_S 1176, Univ. Paris-Sud, Université Paris-Saclay, 94276 Le Kremlin-Bicêtre, France.
authors.forename: Cécile V
authors.initials: CV
authors.lastname: Denis
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D005818:Genetic Engineering; D006487:Hemostasis / Q000235:genetics; D006801:Humans; D007249:Inflammation / Q000235:genetics / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000097:blood / Q000737:chemistry / Q000235:genetics; D010973:Platelet Adhesiveness; D010976:Platelet Count; D056728:von Willebrand Disease, Type 2 / Q000097:blood* / Q000235:genetics / Q000473:pathology*; D014841:von Willebrand Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D050505:Mutant Proteins; D014841:von Willebrand Factor