Ultrahigh-Resolution NMR with Water Signal Suppression for a Deeper Understanding of the Action of Antimetabolic Drugs on Diffuse Large B-Cell Lymphoma
Ultrahigh-Resolution NMR with Water Signal Suppression for a Deeper Understanding of the Action of Antimetabolic Drugs on Diffuse Large B-Cell Lymphoma
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DOI：
10.1021/acs.jproteome.1c00914
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发表时间：
2022-04-01
影响因子：
4.4
通讯作者：
Giraud, Nicolas
中科院分区：
文献类型：
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作者：
Bertho, Gildas;Lordello, Leonardo;Giraud, Nicolas
超高分辨核磁共振近年来在复杂样品分析领域引起了相当大的关注。实际上，宽带同核去耦合技术的实施使我们能够极大地简化拥挤的~1H谱，从所分析的分子中几乎产生每个质子位置的单重态。纯移位方法特别适用于生物样品中代谢物混合物的破译。在这里，我们成功地实现了一个新的纯移位脉冲序列的基础上，心理方法，其中包括一个适合代谢组学分析的溶剂抑制块。由此产生的实验使我们能够记录具有水信号抑制的生物流体的超高分辨率1D NOESY1H光谱，这是突出水相中代谢物混合物的关键一步。我们成功地在弥漫性大B细胞淋巴瘤(DLBCL)细胞外介质上记录了纯移位光谱。尽管灵敏度较低，但纯移位数据的分辨率被发现比标准方法更好，标准方法提供了外源代谢组更详细的愿景。统计分析表明，由此产生的代谢谱使我们能够成功地突出受这些药物影响的几种代谢途径。值得注意的是，我们发现Kidroase在该DLBCL细胞系的代谢途径中起主要作用。
Ultrahigh-resolution NMR has recently attractedconsiderable attention in thefield of complex samples analysis.Indeed, the implementation of broadband homonuclear decou-pling techniques has allowed us to greatly simplify crowded1Hspectra, yielding singlets for almost every proton site from theanalyzed molecules. Pure shift methods have notably shown to beparticularly suitable for deciphering mixtures of metabolites inbiological samples. Here, we have successfully implemented a newpure shift pulse sequence based on the PSYCHE method, whichincorporates a block for solvent suppression that is suitable formetabolomics analysis. The resulting experiment allows us torecord ultrahigh-resolution 1D NOESY1H spectra of biofluidswith suppression of the water signal, which is a crucial step forhighlighting metabolite mixtures in an aqueous phase. We have successfully recorded pure shift spectra on extracellular media ofdiffuse large B-cell lymphoma (DLBCL) cells. Despite a lower sensitivity, the resolution of pure shift data was found to be betterthan that of the standard approach, which provides a more detailed vision of the exo-metabolome. The statistical analyses carried outon the resulting metabolic profiles allow us to successfully highlight several metabolic pathways affected by these drugs. Notably, weshow that Kidrolase plays a major role in the metabolic pathways of this DLBCL cell line