Type 2B von Willebrand disease (VWD) is caused by gain-of-function mutations in von Willebrand factor (VWF). Increased VWF affinity for GPIbα results in loss of high molecular weight multimers and enhanced platelet clearance, both contributing to the bleeding phenotype. Severity of the symptoms vary among type 2B VWD patients, with some developing thrombocytopenia only under stress conditions. Efforts have been made to study underlying pathophysiology for platelet abnormalities, but animal studies have been limited because of species specificity in the VWF-GPIbα interaction. Here, we generated a severe form of type 2B VWD (p.V1316M) knock-in mice in the context of human VWF exon 28 (encoding A1 and A2 domains) and crossed them with human GPIbα transgenic strain. Heterozygous mutant mice recapitulated the phenotype of type 2B VWD in autosomal dominant manner and presented severe macrothrombocytopenia. Of note, platelets remaining in the circulation had extracytoplasmic GPIbα shed off from cell surface. Reciprocal bone marrow transplantation determined mutant VWF produced from endothelial cells as the major cause of platelet phenotype in type 2B VWD mice. Moreover, altered megakaryocyte (MK) maturation in the bone marrow and enhanced extramedullary megakaryopoiesis in the spleen were observed. Interestingly, injection of anti-VWF A1 blocking antibody (NMC-4) not only ameliorated platelet count and GPIbα expression, but also reversed MK ploidy shift. In conclusion, we present a type 2B VWD mouse model with humanized VWF-GPIbα interaction which demonstrated direct influence of aberrant VWFGPIbα binding on MKs.
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