AGK Potentiates Arterial Thrombosis by Affecting Talin-1 and αIIbβ3-Mediated Bidirectional Signaling Pathway
AGK Potentiates Arterial Thrombosis by Affecting Talin-1 and αIIbβ3-Mediated Bidirectional Signaling Pathway
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DOI：
10.1161/atvbaha.122.318647
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发表时间：
2023-04
期刊：
影响因子：
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通讯作者：
Peng Zhang;Haojie Jiang;Mina Yang;Changlong Bi;Kandi Zhang;Dongsheng Liu;M. Wei;Zheyi Jiang;Keyu Lv;C. Fang;Junling Liu;Tiantian Zhang;Yanyan Xu;Junfeng Zhang
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作者：
Peng Zhang;Haojie Jiang;Mina Yang;Changlong Bi;Kandi Zhang;Dongsheng Liu;M. Wei;Zheyi Jiang;Keyu Lv;C. Fang;Junling Liu;Tiantian Zhang;Yanyan Xu;Junfeng Zhang
背景：AGK（酰基甘油激酶）是一种线粒体跨膜蛋白，具有脂质激酶功能。近期研究证实，AGK可促进肿瘤生长和转移，增强CD8+ T细胞糖酵解代谢和功能适应度，或调节巨核细胞分化。然而，AGK在血小板活化和动脉血栓形成中的作用仍有待阐明。方法：采用全自动血液学分析仪进行血液学分析，透射电镜观察血小板形态。我们利用转基因小鼠、体外血小板功能实验和体内血栓形成模型探讨AGK在血小板活化和动脉血栓形成中的作用。通过免疫共沉淀法、质谱法、免疫荧光法和Western blot检测AGK对Talin-1的调控作用。我们通过特异性Elisa试剂盒检测AGK对磷脂酸/溶血磷脂酸脂质合成和凝血酶生成的作用。结果：在本研究中，我们发现AGK缺失或AGK突变对血小板平均体积、血小板微结构和主要血小板膜受体的表达水平没有影响。然而，AGK缺乏或AGK突变显著降低了血小板活化的多个方面，包括激动剂诱导的血小板聚集、颗粒分泌、JON/A结合、在Fg（纤维蛋白原）上扩散和凝块缩回。AGK缺乏或突变可明显延缓动脉血栓形成，但对尾出血时间和血小板促凝功能无影响。机制研究表明，AGK可能促进Talin-1Ser425磷酸化，影响α iib - β3介导的双向信号通路。然而，AGK不影响血小板中磷脂酸/溶血磷脂酸的脂质合成。结论：AGK通过其激酶活性，促进Talin-1 Ser425磷酸化，影响α iib - β3介导的双向信号通路，从而增强血小板活化和动脉血栓形成。
Background: AGK (acylglycerol kinase) was first identified as a mitochondrial transmembrane protein that exhibits a lipid kinase function. Recent studies have established that AGK promotes cancer growth and metastasis, enhances glycolytic metabolism and function fitness of CD8+ T cells, or regulates megakaryocyte differentiation. However, the role of AGK in platelet activation and arterial thrombosis remains to be elaborated. Methods: We performed hematologic analysis using automated hematology analyzer and investigated platelets morphology by transmission electron microscope. We explored the role of AGK in platelet activation and arterial thrombosis utilizing transgenic mice, platelet functional experiments in vitro, and thrombosis models in vivo. We revealed the regulation effect of AGK on Talin-1 by coimmunoprecipitation, mass spectrometry, immunofluorescence, and Western blot. We tested the role of AGK on lipid synthesis of phosphatidic acid/lysophosphatidic acid and thrombin generation by specific Elisa kits. Results: In this study, we found that AGK depletion or AGK mutation had no effect on the platelet average volumes, the platelet microstructures, or the expression levels of the major platelet membrane receptors. However, AGK deficiency or AGK mutation conspicuously decreased multiple aspects of platelet activation, including agonists-induced platelet aggregation, granules secretion, JON/A binding, spreading on Fg (fibrinogen), and clot retraction. AGK deficiency or AGK mutation also obviously delayed arterial thrombus formation but had no effect on tail bleeding time and platelet procoagulant function. Mechanistic investigation revealed that AGK may promote Talin-1Ser425 phosphorylation and affect the αIIbβ3-mediated bidirectional signaling pathway. However, AGK does not affect lipid synthesis of phosphatidic acid/lysophosphatidic acid in platelets. Conclusions: AGK, through its kinase activity, potentiates platelet activation and arterial thrombosis by promoting Talin-1 Ser425 phosphorylation and affecting the αIIbβ3-mediated bidirectional signaling pathway.