Roles of KLF4 and AMPK in the inhibition of glycolysis by pulsatile shear stress in endothelial cells
Roles of KLF4 and AMPK in the inhibition of glycolysis by pulsatile shear stress in endothelial cells
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DOI：
10.1073/pnas.2103982118
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发表时间：
2021-05-25
影响因子：
11.1
通讯作者：
Chien, Shu
中科院分区：
文献类型：
--
作者：
Han, Yue;He, Ming;Chien, Shu
血管内皮细胞（EC）感知并响应血流动力学力，如脉动剪切应力（PS）和振荡剪切应力（OS）。在代谢途径中，糖酵解受动脉粥样硬化酮OS和动脉粥样硬化保护PS的差异调节。通过研究PS在表观遗传学、转录组学和激酶组学水平抑制糖酵解通量的分子机制，我们已经证明了葡萄糖激酶调节蛋白（GCKR）在体外和体内都被PS显著诱导，尽管PS下调其他糖酵解酶如己糖激酶（HK 1）。使用下一代测序数据，我们确定了PS诱导的Kruppel样因子4（KLF 4）的结合，其作为先锋转录因子，与GCKR启动子结合以改变GCKR反式激活的染色质结构。在翻译后水平，PS激活的AMP激活的蛋白激酶（AMPK）磷酸化GCKR的Ser-481，从而增强GCKR和HK 1之间的相互作用在EC。在体内，与EC消融AMPK α 2（AMPK α 2（-/-））的同窝小鼠相比，野生型AMPK α 2（+/+）小鼠胸主动脉中磷酸化GCKR Ser-481的水平以及GCKR与HK 1之间的相互作用增加。此外，高水平自愿跑轮的小鼠的睾丸中GCKR水平升高。PS诱导GCKR的潜在机制涉及KLF 4在表观遗传水平和AMPK在翻译后水平的调节。
Vascular endothelial cells (ECs) sense and respond to hemodynamic forces such as pulsatile shear stress (PS) and oscillatory shear stress (OS). Among the metabolic pathways, glycolysis is differentially regulated by atheroprone OS and atheroprotective PS. Studying the molecular mechanisms by which PS suppresses glycolytic flux at the epigenetic, transcriptomic, and kinomic levels, we have demonstrated that glucokinase regulatory protein (GCKR) was markedly induced by PS in vitro and in vivo, although PS down-regulates other glycolysis enzymes such as hexokinase (HK1). Using next-generation sequencing data, we identified the binding of PS-induced Kruppel-like factor 4 (KLF4), which functions as a pioneer transcription factor, binding to the GCKR promoter to change the chromatin structure for transactivation of GCKR. At the posttranslational level, PS-activated AMP-activated protein kinase (AMPK) phosphorylates GCKR at Ser-481, thereby enhancing the interaction between GCKR and HK1 in ECs. In vivo, the level of phosphorylated GCKR Ser-481 and the interaction between GCKR and HK1 were increased in the thoracic aorta of wild-type AMPK alpha 2(+/+) mice in comparison with littermates with EC ablation of AMPK alpha 2 (AMPK alpha 2(-/-)). In addition, the level of GCKR was elevated in the aortas of mice with a high level of voluntary wheel running. The underlying mechanisms for the PS induction of GCKR involve regulation at the epigenetic level by KLF4 and at the post-translational level by AMPK.