Qiao, JianlinWu, XiaoqingLuo, QiWei, GuangyuXu, MengdiWu, YuluLiu, YunLi, XiaoqianZi, JieJu, WenGardiner, Elizabeth2019-07-292019-07-290390-6078http://hdl.handle.net/1885/164779In addition to their hemostatic function, platelets play an important role in regulating the inflammatory response. The platelet NLRP3 inflammasome not only promotes interleukin-1β secretion, but was also found to be upregulated during platelet activation and thrombus formation in vitro. However, the role of NLRP3 in platelet function and thrombus formation in vivo remains unclear. In this study, we aimed to investigate the role of NLRP3 in platelet integrin aIIbβ3 signaling transduction. Using NLRP3-/- mice, we showed that NLRP3-deficient platelets do not have significant differences in expression of the platelet-specific adhesive receptors aIIbβ3 integrin, GPIba or GPVI; however, NLRP3-/-platelets transfused into wild-type mice resulted in prolonged tail-bleed-ing time and delayed arterial thrombus formation, as well as exhibiting impaired spreading on immobilized fibrinogen and defective clot retraction, concomitant with decreased phosphorylation of c-Src, Syk and PLCγ2 in response to thrombin stimulation. Interestingly, addition of exogenous recombinant interleukin-1β reversed the defect in NLRP3-/-platelet spreading and clot retraction, and restored thrombin-induced phosphorylation of c-Src/Syk/PLCγ2, whereas an anti-interleukin-1β antibody blocked spreading and clot retraction mediated by wild-type platelets. Using the direct NLRP3 inhibitor, CY-09, we demonstrated significantly reduced human platelet aggregation in response to threshold concentrations of collagen and ADP, as well as impaired clot retraction in CY-09-treated human platelets, supporting a role for NLRP3 also in regulating human platelet aIIbβ3 outside-in signaling. This study identifies a novel role for NLRP3 and interleukin-1β in platelet function, and provides a new potential link between thrombosis and inflammation, suggesting that therapies targeting NLRP3 or interleukin-1β might be beneficial for treating inflammation-associated thrombosis.This research was supported by the National Natural Science Foundation of China (grant n. 81400082, 81370602, 81570096, 81671584, 81641151 and 81700178), the Natural Science Foundation of Jiangsu Province (grant n. BK20141138 BK20140219), funding for the Distinguished Professorship Program of Jiangsu Province, the Shuangchuang Project of Jiangsu Province, the National Health and Medical Research Council of Australia, the Six Talent Peaks Project of Jiangsu Province (WSN-133), the 333 projects of Jiangsu Province (BRA2017542), the Key University Science Research Project of Jiangsu Province (17KJA320008), Jiangsu Province’s Key Provincial Talents Program (ZDRCA2016054), the Colleges’ Science Foundation of Jiangsu Province (16KJB320013), Postgraduate Research Innovation Project of Jiangsu Province (KYCX18_2186), and Key University Science Research Project of Jiangsu Province (18KJA320010).application/pdfen-AU© 2018 Ferrata Storti Foundationhttps://creativecommons.org/licenses/by-nc/4.0/legalcode201810.3324/haematol.2018.1917002019-03-31Creative Commons Attribution-NonCommercial 4.0 International