Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan.
Healthy Aging Research Center, Chang Gung University, Taoyuan 33302, Taiwan.
Cells. 2020 Feb 18;9(2):473. doi: 10.3390/cells9020473.
Enterovirus 71 (EV71) infection is an endemic disease in Southeast Asia and China. We have previously shown that EV71 virus causes functional changes in mitochondria. It is speculative whether EV71 virus alters the host cell metabolism to its own benefit. Using a metabolomics approach, we demonstrate that EV71-infected Vero cells had significant changes in metabolism. Glutathione and its related metabolites, and several amino acids, such as glutamate and aspartate, changed significantly with the infectious dose of virus. Other pathways, including glycolysis and tricarboxylic acid cycle, were also altered. A change in glutamine/glutamate metabolism is critical to the viral infection. The presence of glutamine in culture medium was associated with an increase in viral replication. Dimethyl α-ketoglutarate treatment partially mimicked the effect of glutamine supplementation. In addition, the immunoblot analysis revealed that the expression of glutamate dehydrogenase (GDH) and trifunctional carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase (CAD) increased during infection. Knockdown of expression of glutaminase (GLS), GDH and CAD drastically reduced the cytopathic effect (CPE) and viral replication. Furthermore, we found that CAD bound VP1 to promote the de novo pyrimidine synthesis. Our findings suggest that virus may induce metabolic reprogramming of host cells to promote its replication through interactions between viral and host cell proteins.
肠道病毒 71 型（EV71）感染是东南亚和中国的地方性疾病。我们之前已经表明，EV71 病毒会导致线粒体功能发生变化。EV71 病毒是否会改变宿主细胞代谢以适应自身需要，这还只是推测。通过代谢组学方法，我们证明 EV71 感染的 Vero 细胞的代谢发生了显著变化。谷胱甘肽及其相关代谢物以及几种氨基酸，如谷氨酸和天冬氨酸，随着病毒感染剂量的变化而显著变化。其他途径，包括糖酵解和三羧酸循环，也发生了改变。谷氨酰胺/谷氨酸代谢的变化对病毒感染至关重要。培养基中存在谷氨酰胺会增加病毒复制。二甲基-α-酮戊二酸处理部分模拟了谷氨酰胺补充的效果。此外，免疫印迹分析显示，谷氨酸脱氢酶（GDH）和三功能氨甲酰磷酸合成酶 2、天冬氨酸转氨甲酰酶和二氢乳清酸酶（CAD）的表达在感染过程中增加。敲低谷氨酰胺酶（GLS）、GDH 和 CAD 的表达水平可显著降低致细胞病变效应（CPE）和病毒复制。此外，我们发现 CAD 与 VP1 结合以促进新的嘧啶合成。我们的研究结果表明，病毒可能通过病毒和宿主细胞蛋白之间的相互作用，诱导宿主细胞的代谢重编程，从而促进其复制。
