Inositol hexakisphosphate kinase 1 maintains hemostasis in mice by regulating platelet polyphosphate levels

Abstract

Polyphosphate (polyP), a polymer of orthophosphate moieties released from the dense granules of activated platelets, is a procoagulant agent. Inositol pyrophosphates, another group of phosphate-rich molecules, consist of mono- and diphosphates substituted on an inositol ring. Diphosphoinositol pentakisphosphate (IP₇), the most abundant inositol pyrophosphate, is synthesized on phosphorylation of inositol hexakisphosphate (IP₆) by IP₆ kinases, of which there are 3 mammalian isoforms (IP6K1/2/3) and a single yeast isoform. Yeast lacking IP₆ kinase are devoid of polyP, suggesting a role for IP₆ kinase in maintaining polyP levels. We theorized that the molecular link between IP₆ kinase and polyP is conserved in mammals and investigated whether polyP-dependent platelet function is altered in IP6K1 knockout (Ip6k1^−/−) mice. We observe a significant reduction in platelet polyP levels in Ip6k1^−/− mice, along with slower platelet aggregation and lengthened plasma clotting time. Incorporation of polyP into fibrin clots was reduced in Ip6k1^−/− mice, thereby altering clot ultrastructure, which was rescued on the addition of exogenous polyP. In vivo assays revealed longer tail bleeding time and resistance to thromboembolism in Ip6k1^−/− mice. Taken together, our data suggest a novel role for IP6K1 in regulation of mammalian hemostasis via its control of platelet polyP levels.