Mitochondrial pyruvate dehydrogenase kinases contribute to platelet function and thrombosis in mice by regulating aerobic glycolysis.
Mitochondrial pyruvate dehydrogenase kinases contribute to platelet function and thrombosis in mice by regulating aerobic glycolysis.
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DOI：
10.1182/bloodadvances.2023010100
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发表时间：
2023-06-13
期刊：
影响因子：
7.5
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中科院分区：
文献类型：
--
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丙酮酸脱氢酶激酶（PDK）2和4的联合缺失抑制血小板功能和动脉血栓形成。PDK 2/4通过调节活化血小板中的丙酮酸脱氢酶磷酸化和有氧糖酵解来促进血小板生物学。静息血小板依赖于氧化磷酸化（OXPHOS）和有氧糖酵解（在氧气存在下葡萄糖转化为乳酸盐）来满足其能量需求。相比之下，血小板活化相对于OXPHOS表现出增加的有氧糖酵解速率。线粒体酶丙酮酸脱氢酶激酶（PDK）磷酸化丙酮酸脱氢酶（PDH）复合物以抑制其活性，从而在血小板活化时将丙酮酸流量从OXPHOS转移到有氧糖酵解。在4种PDK亚型中，PDK 2和PDK 4（PDK 2/4）主要与代谢性疾病相关。在此，我们报告了PDK 2/4的联合缺失抑制激动剂诱导的血小板功能，包括聚集、整合素αIIbβ3活化、脱粒、扩散和凝块收缩。此外，胶原介导的PLCγ2磷酸化和钙动员在PDK 2/4−/−血小板中显著减少，表明GPVI信号传导受损。PDK 2/4−/−小鼠对FeCl 3诱导的颈动脉和激光诱导的肠系膜动脉血栓形成不太敏感，对止血没有任何影响。在过继转移实验中，与输注WT血小板的hIL-4 R α/GPIbα-Tg小鼠相比，输注PDK 2/4−/−血小板的血小板减少性hIL-4 R α/GPIbα转基因小鼠对FeCl 3损伤诱导的颈动脉血栓形成的易感性较低，表明PDK 2/4在血栓形成中具有血小板特异性作用。从机制上讲，PDK 2/4缺失对血小板功能的抑制作用与活化血小板中PDH磷酸化和glycoPER的降低有关，表明PDK 2/4调节有氧糖酵解。最后，使用PDK 2或PDK 4单基因敲除小鼠，我们鉴定了PDK 4在调节血小板分泌和血栓形成中比PDK 2发挥更突出的作用。本研究确定了PDK 2/4在调节血小板功能中的基本作用，并确定了PDK/PDH轴作为潜在的新型抗血栓形成靶点。
Combined deletion of pyruvate dehydrogenase kinase (PDK) 2 and 4 inhibits platelet function and arterial thrombosis. PDK2/4 contributes to platelet biology by regulating pyruvate dehydrogenase phosphorylation and aerobic glycolysis in activated platelets. Resting platelets rely on oxidative phosphorylation (OXPHOS) and aerobic glycolysis (conversion of glucose to lactate in the presence of oxygen) for their energy requirements. In contrast, platelet activation exhibits an increased rate of aerobic glycolysis relative to OXPHOS. Mitochondrial enzymes pyruvate dehydrogenase kinases (PDKs) phosphorylate the pyruvate dehydrogenase (PDH) complex to inhibit its activity, thereby diverting the pyruvate flux from OXPHOS to aerobic glycolysis upon platelet activation. Of 4 PDK isoforms, PDK2 and PDK4 (PDK2/4) are predominantly associated with metabolic diseases. Herein, we report that the combined deletion of PDK2/4 inhibits agonist-induced platelet functions, including aggregation, integrin αIIbβ3 activation, degranulation, spreading, and clot retraction. In addition, collagen-mediated PLCγ2 phosphorylation and calcium mobilization were significantly reduced in PDK2/4−/− platelets, suggesting impaired GPVI signaling. The PDK2/4−/− mice were less susceptible to FeCl3-induced carotid and laser-induced mesenteric artery thrombosis without any effect on hemostasis. In adoptive transfer experiments, thrombocytopenic hIL-4Rα/GPIbα-transgenic mice transfused with PDK2/4−/− platelets exhibited less susceptibility to FeCl3 injury–induced carotid thrombosis compared with hIL-4Rα/GPIbα-Tg mice transfused with WT platelets, suggesting a platelet-specific role of PDK2/4 in thrombosis. Mechanistically, the inhibitory effects of PDK2/4 deletion on platelet function were associated with reduced PDH phosphorylation and glycoPER in activated platelets, suggesting that PDK2/4 regulates aerobic glycolysis. Finally, using PDK2 or PDK4 single KO mice, we identified that PDK4 plays a more prominent role in regulating platelet secretion and thrombosis compared with PDK2. This study identifies the fundamental role of PDK2/4 in regulating platelet functions and identifies the PDK/PDH axis as a potentially novel antithrombotic target.
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8.7
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