201. Heterozygous ITGA2B R995W mutation inducing constitutive activation of the αIIbβ3 receptor affects proplatelet formation and causes congenital macrothrombocytopenia.
- Author
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Kunishima S, Kashiwagi H, Otsu M, Takayama N, Eto K, Onodera M, Miyajima Y, Takamatsu Y, Suzumiya J, Matsubara K, Tomiyama Y, and Saito H
- Subjects
- Adult, Amino Acid Substitution, Animals, CHO Cells, Cell Line, Child, Child, Preschool, Cricetinae, Cricetulus, DNA Mutational Analysis, Female, Heterozygote, Humans, Infant, Integrin alpha2 chemistry, Integrin alpha2 metabolism, Integrin beta3 genetics, Integrin beta3 metabolism, Male, Megakaryocytes metabolism, Megakaryocytes pathology, Mice, Middle Aged, Mutant Proteins chemistry, Mutant Proteins genetics, Mutant Proteins metabolism, Mutation, Missense, Protein Conformation, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Thrombocytopenia blood, Transfection, Young Adult, Integrin alpha2 genetics, Platelet Glycoprotein GPIIb-IIIa Complex metabolism, Thrombocytopenia congenital, Thrombocytopenia genetics
- Abstract
Congenital macrothrombocytopenia is a genetically heterogeneous group of rare disorders. αIIbβ3 has not been implicated in these conditions. We identified a novel, conserved heterozygous ITGA2B R995W mutation in 4 unrelated families. The surface expression of platelet αIIbβ3 was decreased to 50% to 70% of control. There was spontaneous PAC-1 and fibrinogen binding to resting platelets without CD62p expression. The activation state of αIIbβ3 in 293T cells was higher for αIIb-W995 than for β3-H723 but was weaker than for β3-N562. FAK was spontaneously phosphorylated in αIIb-W995/β3-transfected 293T cells. These results indicate that αIIb-W995/β3 has a constitutive, activated conformation but does not induce platelet activation. αIIb-W995/β3-transfected CHO cells developed membrane ruffling and abnormal cytoplasmic protrusions. The increased size and decreased number of proplatelet tips in αIIb-W995/β3-transduced mouse fetal liver-derived megakaryocytes indicate defective pro-platelet formation. We propose that activating mutations in ITGA2B and ITGB3 represent the etiology of a subset of congenital macrothrombocytopenias.
- Published
- 2011
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