Autophagy is important for maintaining cellular homeostasis, and thus its deficiency is implicated in a broad spectrum of human diseases. Its role in platelet function has only recently been examined. Our biochemical and imaging studies demonstrate that the core autophagy machinery exists in platelets, and that autophagy is constitutively active in resting platelets. Moreover, autophagy is induced upon platelet activation, as indicated by agonist-induced loss of the autophagy marker LC3II. Additional experiments, using inhibitors of platelet activation, proteases, and lysosomal acidification, as well as platelets from knockout mouse strains, show that agonist-induced LC3II loss is a consequence of platelet signaling cascades and requires proteases, acidic compartments, and membrane fusion. To assess the physiological role of platelet autophagy, we generated a mouse strain with a megakaryocyte- and platelet-specific deletion of Atg7, an enzyme required for LC3II production. Ex vivo analysis of platelets from these mice shows modest defects in aggregation and granule cargo packaging. Although these mice have normal platelet numbers and size distributions, they exhibit a robust bleeding diathesis in the tail-bleeding assay and a prolonged occlusion time in the FeCl3-induced carotid injury model. Our results demonstrate that autophagy occurs in platelets and is important for hemostasis and thrombosis.
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This work was supported by National Institutes of Health Grants P20GM103486 (National Institute of General Medical Sciences; to Q.J.W.), HL56652 and HL091893 (National Heart, Lung, and Blood Institute; to S.W.W.), R01NS060123 (National Institute of Neurological Disorders; to Z.Y.), and R01HL119393 (National Heart, Lung, and Blood Institute; to B.S.); the Ellison Medical Foundation (Q.J.W.); University of Kentucky Research Support Grant and start-up fund (Q.J.W.); and American Heart Association Predoctoral Fellowship (Y.H.).
The online version of this article contains a data supplement.
Ouseph, Madhu M.; Huang, Yunjie; Banerjee, Meenakshi; Joshi, Smita; MacDonald, Laura; Zhong, Yu; Liu, Huijuan; Li, Xianting; Xiang, Binggang; Zhang, Guoying; Komatsu, Masaaki; Yue, Zhenyu; Li, Zhenyu; Storrie, Brian; Whiteheart, Sidney W.; and Wang, Qing Jun, "Autophagy Is Induced Upon Platelet Activation and Is Essential for Hemostasis and Thrombosis" (2015). Molecular and Cellular Biochemistry Faculty Publications. 149.