Humanized GPIbα–von Willebrand factor interaction in the mouse
| Autor: | Yu-ichi Kamikubo, Rashmi Sood, Yosuke Morodomi, Stefano Forli, Jennifer N. Orje, Robert R. Montgomery, Zaverio M. Ruggeri, Sachiko Kanaji, Sandra L. Haberichter, Alessandro Zarpellon, Yunfeng Chen, Jerome Eberhardt, Taisuke Kanaji, Scot A. Fahs |
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| Rok vydání: | 2018 |
| Předmět: |
Blood Platelets
0301 basic medicine Genetically modified mouse Protein Conformation animal diseases Transgene Mice Transgenic Plasma protein binding Molecular Dynamics Simulation 030204 cardiovascular system & hematology Platelet membrane glycoprotein Thrombosis and Hemostasis Mice Protein Aggregates Structure-Activity Relationship 03 medical and health sciences 0302 clinical medicine Von Willebrand factor Bleeding time hemic and lymphatic diseases von Willebrand Factor medicine Animals Humans Platelet Crosses Genetic Hemostasis biology medicine.diagnostic_test Chemistry Thrombosis Exons Hematology Surface Plasmon Resonance Molecular biology Molecular Docking Simulation 030104 developmental biology Platelet Glycoprotein GPIb-IX Complex cardiovascular system biology.protein Protein Multimerization Biomarkers circulatory and respiratory physiology Protein Binding |
| Zdroj: | Blood Advances. 2:2522-2532 |
| ISSN: | 2473-9537 2473-9529 |
| DOI: | 10.1182/bloodadvances.2018023507 |
| Popis: | The interaction of platelet glycoprotein Ibα (GPIbα) with von Willebrand factor (VWF) initiates hemostasis after vascular injury and also contributes to pathological thrombosis. GPIbα binding to the VWF A1 domain (VWFA1) is a target for antithrombotic intervention, but attempts to develop pharmacologic inhibitors have been hindered by the lack of animal models because of the species specificity of the interaction. To address this problem, we generated a knockin mouse with Vwf exon 28–encoding domains A1 and A2 replaced by the human homolog (VWFh28). VWFh28 mice (M1HA) were crossbred with a transgenic mouse strain expressing human GPIbα on platelets (mGPIbαnull;hGPIbαTg; H1MA) to generate a new strain (H1HA) with humanized GPIbα-VWFA1 binding. Plasma VWF levels in the latter 3 strains were similar to those of wild-type mice (M1MA). Compared with the strains that had homospecific GPIbα-VWF pairing (M1MA and H1HA), M1HA mice of those with heterospecific pairing had a markedly greater prolongation of tail bleeding time and attenuation of thrombogenesis after injury to the carotid artery than H1MA mice. Measurements of GPIbα-VWFA1 binding affinity by surface plasmon resonance agreed with the extent of observed functional defects. Ristocetin-induced platelet aggregation was similar in H1HA mouse and human platelet-rich plasma, and it was comparably inhibited by monoclonal antibody NMC-4, which is known to block human GPIbα-VWFA1 binding, which also inhibited FeCl3-induced mouse carotid artery thrombosis. Thus, the H1HA mouse strain is a fully humanized model of platelet GPIbα-VWFA1 binding that provides mechanistic and pharmacologic information relevant to human hemostatic and thrombotic disorders. |
| Databáze: | OpenAIRE |
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