Gao Xia, Lin Ling, Hu Anqi, Zhao Heyu, Kang Le, Wang Xiaoyu, Yuan Chunyan, Yang Pengyuan, Shen Huali
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China.
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China; Xiamen Cardiovascular Hospital, Xiamen University, Xiamen, 361004, Fujian, China.
Talanta. 2022 Aug 1;245:123475. doi: 10.1016/j.talanta.2022.123475. Epub 2022 Apr 12.
Sphingolipids (SPLs) are bioactive lipids that manifest structural diversity and complexity in eukaryotes. However, the distributions and functions of these molecules in mammalian tissues/cells have not been systematically investigated. Herein, we integrated shotgun lipidomics with targeted LC-MRM/MS approach to comprehensively analyze SPL species in various biological samples with high accuracy. Preliminarily, 1311 SPL molecules were identified in 18 kinds of mammalian samples, including 3 groups of human sera, 10 mouse tissues and 5 cell lines via 26 sphingoid long-chain bases scanning. The sphingolipidome compositions and distributions were systematically characterized and distinct qualitative and quantitative profiles were clearly exhibited in various samples, indicating unique biological functions of the sphingolipidomes. Next, targeted SPLs analysis by LC-MRM/MS with critical criteria monitoring two characteristic fragments of one precursor was applied to human serum samples from 24 coronary artery disease (CAD) patients and 12 healthy controls, which successfully quantified 170 SPL molecules. Ten novel SPL molecules were discovered as a potential diagnostic panel for CAD patients via multivariate exploratory receiver operating characteristic curve-based biomarker analysis. The diagnostic panel with the 10 SPL molecules achieved 97.2% accuracy, with a favorable auxiliary diagnostic value (AUC = 1.000), for the detection of CAD. These results clearly support the sphingolipidomic approach in application to discovering disease biomarker panel as well as deep investigation of biological functions of complex SPLs in mammalian samples.
鞘脂（SPLs）是一类生物活性脂质，在真核生物中表现出结构的多样性和复杂性。然而，这些分子在哺乳动物组织/细胞中的分布和功能尚未得到系统研究。在此，我们将鸟枪法脂质组学与靶向液相色谱-多反应监测/质谱方法相结合，以高精度全面分析各种生物样品中的鞘脂种类。初步通过26种鞘氨醇长链碱基扫描，在18种哺乳动物样品中鉴定出1311种鞘脂分子，包括3组人血清、10种小鼠组织和5种细胞系。系统地表征了鞘脂组的组成和分布，不同样品中清晰呈现出独特的定性和定量特征，表明鞘脂组具有独特的生物学功能。接下来，采用基于液相色谱-多反应监测/质谱的靶向鞘脂分析方法，以监测一种前体的两个特征性片段的关键标准，对来自24例冠状动脉疾病（CAD）患者和12例健康对照的人血清样品进行分析，成功定量了170种鞘脂分子。通过基于多变量探索性受试者工作特征曲线的生物标志物分析，发现了10种新型鞘脂分子作为CAD患者的潜在诊断指标。包含这10种鞘脂分子的诊断指标在检测CAD时的准确率达到97.2%，具有良好的辅助诊断价值（AUC = 1.000）。这些结果明确支持了鞘脂组学方法在发现疾病生物标志物指标以及深入研究哺乳动物样品中复杂鞘脂生物学功能方面的应用。
