Disease Biology Lab, SSSIHL-Agilent Center for Excellence in Multiomics and Cell Sciences, Dept. of Biosciences, Sri Sathya Sai Institute of Higher Learning, Prasanthi Nilayam, Anantapur, Andhra Pradesh, 515 134, India.
Dept. of Botany/Biotechnology, CMS College, Kottayam, 686 001, India.
Sci Rep. 2021 May 7;11(1):9766. doi: 10.1038/s41598-021-89137-z.
Glaucoma of which primary open angle glaucoma (POAG) constitutes 75%, is the second leading cause of blindness. Elevated intra ocular pressure and Nitric oxide synthase (NOS) dysfunction are hallmarks of POAG. We analyzed clinical data, cytokine profile, ATP level, metabolomics and GEO datasets to identify features unique to POAG. N9 microglial cells are used to gain mechanistic insights. Our POAG cohort showed elevated ATP in aqueous humor and cytokines in plasma. Metabolomic analysis showed changes in 21 metabolites including Dimethylarginine (DMAG) and activation of tryptophan metabolism in POAG. Analysis of GEO data sets and previously published proteomic data sets bins genes into signaling and metabolic pathways. Pathways from reanalyzed metabolomic data from literature significantly overlapped with those from our POAG data. DMAG modulated purinergic signaling, ATP secretion and cytokine expression were inhibited by N-Ethylmaleimide, NO donors, BAPTA and purinergic receptor inhibitors. ATP induced elevated intracellular calcium level and cytokines expression were inhibited by BAPTA. Metabolomics of cell culture supernatant from ATP treated sets showed metabolic deregulation and activation of tryptophan metabolism. DMAG and ATP induced IDO1/2 and TDO2 were inhibited by N-Ethylmaleimide, sodium nitroprusside and BAPTA. Our data obtained from clinical samples and cell culture studies reveal a strong association of elevated DMAG, ATP, cytokines and activation of tryptophan metabolism with POAG. DMAG mediated ATP signaling, inflammation and metabolic remodeling in microglia might have implications in management of POAG.
青光眼是全球第二大致盲性眼病,其中原发性开角型青光眼(POAG)占 75%。眼内压升高和一氧化氮合酶(NOS)功能障碍是 POAG 的主要特征。我们分析了临床数据、细胞因子谱、ATP 水平、代谢组学和 GEO 数据集,以确定 POAG 特有的特征。我们使用 N9 小胶质细胞来获得机制见解。我们的 POAG 队列显示房水中 ATP 和血浆中的细胞因子升高。代谢组学分析显示包括二甲基精氨酸(DMAG)在内的 21 种代谢物发生变化,并激活了 POAG 中的色氨酸代谢。对 GEO 数据集和以前发表的蛋白质组学数据集的分析将基因分类为信号转导和代谢途径。重新分析来自文献的代谢组学数据的途径与我们的 POAG 数据的途径显著重叠。DMAG 调节嘌呤能信号转导,N-乙基马来酰亚胺、NO 供体、BAPTA 和嘌呤能受体抑制剂抑制 ATP 分泌和细胞因子表达。ATP 诱导的细胞内钙离子水平升高和细胞因子表达被 BAPTA 抑制。用 ATP 处理的细胞培养上清代谢组学显示代谢失调和色氨酸代谢的激活。N-乙基马来酰亚胺、硝普钠和 BAPTA 抑制 DMAG 和 ATP 诱导的 IDO1/2 和 TDO2。我们从临床样本和细胞培养研究中获得的数据揭示了升高的 DMAG、ATP、细胞因子和色氨酸代谢激活与 POAG 之间的强烈关联。DMAG 介导的 ATP 信号转导、炎症和小胶质细胞的代谢重塑可能对 POAG 的管理具有重要意义。