Metabolites Interrogation in Cell Fate Decision of Cultured Human Corneal Endothelial Cells
Metabolites Interrogation in Cell Fate Decision of Cultured Human Corneal Endothelial Cells
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DOI：
10.1167/iovs.61.2.10
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发表时间：
2020-02-01
影响因子：
4.4
通讯作者：
Sotozono, Chie
中科院分区：
文献类型：
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作者：
Hamuro, Junji;Numa, Kohsaku;Sotozono, Chie
目的。旨在阐明培养的人角膜内皮细胞（cHCEC）细胞命运决定中的代谢问题。方法。为了分析培养上清液 (CS) 中的代谢物，通过设施服务对成熟分化和具有细胞状态转变的各种 cHCEC 进行了 34 项代谢组测量。通过液相色谱串联质谱 (LC-MS/MS) 对每个高质量或低质量 cHCEC 亚群 (SP) 的 3 等分试样进行细胞裂解物的综合蛋白质组学研究。使用 DAVID 及其选项“KEGG_PATHWAY”对参与 cHCEC 代谢的重点基因进行了研究。结果。代谢物簇与 CD44-/+ SP 的独特含量非常吻合。 CS中分泌的丙酮酸和乳酸均与优质SP的含量呈负相关。 CS中乳酸、丙酮酸与Ile、Leu、Ser呈正相关，与谷氨酰胺呈负相关。 CS中的血小板源性生长因子-β与CS中的乳酸呈负相关，间接表明与CD44-/+ SPs的含量呈正相关。糖酵解酶的上调和谷氨酰胺流入三羧酸循环可能与代谢重新布线有关，该代谢重新布线将成熟分化的 CD44-/+PSP 中的氧化代谢转化为具有细胞状态转变的未成熟 SP 中的糖酵解通量依赖性状态。结论。研究结果表明，cHCEC 的细胞命运决定可能至少部分是通过代谢重连决定的。
PURPOSE. Aiming to clarify the metabolic interrogation in cell fate decision of cultured human corneal endothelial cells (cHCECs).METHODS. To analyze the metabolites in the culture supernatants (CS), 34 metabolome measurements were carried out for mature differentiated and a variety of cHCECs with cell state transition through a facility service. Integrated proteomics research for cell lysates by liquid chromatography tandem mass spectrometry (LC-MS/MS) was performed for 3 aliquots of each high-quality or low-quality cHCEC subpopulations (SP). The investigations for the focused genes involved in cHCEC metabolism were performed by using DAVID and its options "KEGG_PATHWAY."RESUETS. The clusters of metabolites coincided well with the distinct content of CD44-/+ SPs. Both secreted pyruvic acid and lactic acid in the CS were negatively correlated with the content of high-quality SPs. Lactic acid and pyruvic acid in the CS exhibited the positive correlation with that of Ile, Leu, and Ser, whereas the negative correlation was with glutamine. Platelet-derived growth factor-beta beta in the CS negatively correlated with lactic acid in CS, indicating indirectly the positive correlation with the content of CD44-/+ SPs. Upregulated glycolytic enzymes and influx of glutamine to the tricarboxylic acid cycle may be linked with a metabolic rewiring converting oxidative metabolism in mature differentiated CD44-/+PSPs into a glycolytic flux-dependent state in immature SPs with cell state transition.CONCLUSIONS. The findings suggest that the cell fate decision of cHCECs may be dictated at least partly through metabolic rewiring.