Gain-of-Function Mutation in Filamin A Potentiates Platelet Integrin αIIbβ3 Activation

Objective-Dominant mutations of the X-linked filamin A (FLNA) gene are responsible for filaminopathies A, which are rare disorders including brain periventricular nodular heterotopia, congenital intestinal pseudo-obstruction, cardiac valves or skeleton malformations, and often macrothrombocytopenia. Approach and Results-We studied a male patient with periventricular nodular heterotopia and congenital intestinal pseudoobstruction, his unique X-linked FLNA allele carrying a stop codon mutation resulting in a 100-amino acid-long FLNa C-terminal extension (NP_001447.2: p.Ter2648SerextTer101). Platelet counts were normal, with few enlarged platelets. FLNa was detectable in all platelets but at 30% of control levels. Surprisingly, all platelet functions were significantly upregulated, including platelet aggregation and secretion, as induced by ADP, collagen, or von Willebrand factor in the presence of ristocetin, as well as thrombus formation in blood flow on a collagen or on a von Willebrand factor matrix. Most importantly, patient platelets stimulated with ADP exhibited a marked increase in alpha(IIb)beta(3) integrin activation and a parallel increase in talin recruitment to beta(3), contrasting with normal Rap1 activation. These results are consistent with the mutant FLNa affecting the last step of alpha(IIb)beta(3) activation. Overexpression of mutant FLNa in the HEL megakaryocytic cell line correlated with an increase (compared with wild-type FLNa) in PMA-induced fibrinogen binding to and in talin and kindlin-3 recruitment by alpha(IIb)beta(3). Conclusions-Altogether, our results are consistent with a less binding of mutant FLNa to beta(3) and the facilitated recruitment of talin by beta(3) on platelet stimulation, explaining the increased alpha(IIb)beta(3) activation and the ensuing gain-of-platelet functions.