背景:糖酵解抑制剂2-脱氧-d-葡萄糖(2-DG)以非竞争性方式结合己糖激酶,以竞争性方式结合磷酸葡萄糖异构酶,阻断糖酵解途径的初始步骤。虽然2-DG刺激内质网(ER)应激,激活未折叠的蛋白质反应以恢复蛋白质稳态,目前尚不清楚哪些ER应激相关基因在人原代细胞中响应2-DG处理而被调节。这里,我们的目的是确定用2-DG处理单核细胞和单核细胞衍生的巨噬细胞(MDMs)是否会导致ER应激特有的转录谱.
方法:我们进行了生物信息学分析,以鉴定先前报道的2-DG处理细胞的RNA-seq数据集中的差异表达基因(DEG)。进行RT-qPCR以验证培养的MDMs上的测序数据。
结果:通过2-DG处理的单核细胞和MDMs的转录分析发现总共95个常见的DEGs。其中,74个上调,21个下调。多转录物分析表明,DEGs与整合的应激反应(GRP78/BiP,PERK,ATF4,CHOP,GADD34,IRE1α,XBP1,SESN2,ASNS,PHGDH),己糖胺生物合成途径(GFAT1,GNA1,PGM3,UAP1),和甘露糖代谢(GMPPA和GMPPB)。
结论:结果表明,2-DG触发了一个基因表达程序,该程序可能参与恢复原代细胞的蛋白质稳态。
结论:已知2-DG抑制糖酵解并诱导ER应激;然而,其对原代细胞基因表达的影响尚不清楚。这项工作表明2-DG是改变单核细胞和巨噬细胞代谢状态的应激诱导剂。
Glycolytic inhibitor 2-deoxy-d-glucose (2-DG) binds to hexokinase in a non-competitive manner and phosphoglucose isomerase in a competitive manner, blocking the initial steps of the glycolytic pathway. Although 2-DG stimulates endoplasmic reticulum (ER) stress, activating the unfolded protein response to restore protein homeostasis, it is unclear which ER stress-related genes are modulated in response to 2-DG treatment in human primary cells. Here, we aimed to determine whether the treatment of monocytes and monocyte-derived macrophages (MDMs) with 2-DG leads to a transcriptional profile specific to ER stress.
We performed bioinformatics analysis to identify differentially expressed genes (DEGs) in previously reported RNA-seq datasets of 2-DG treated cells. RT-qPCR was performed to verify the sequencing data on cultured MDMs.
A total of 95 common DEGs were found by transcriptional analysis of monocytes and MDMs treated with 2-DG. Among these, 74 were up-regulated and 21 were down-regulated. Multitranscript analysis showed that DEGs are linked to integrated stress response (GRP78/BiP, PERK, ATF4, CHOP, GADD34, IRE1α, XBP1, SESN2, ASNS, PHGDH), hexosamine biosynthetic pathway (GFAT1, GNA1, PGM3, UAP1), and mannose metabolism (GMPPA and GMPPB).
Results reveal that 2-DG triggers a gene expression program that might be involved in restoring protein homeostasis in primary cells.
2-DG is known to inhibit glycolysis and induce ER stress; however, its effect on gene expression in primary cells is not well understood. This work shows that 2-DG is a stress inducer shifting the metabolic state of monocytes and macrophages.