Isomer-specific profiling of N-glycans derived from human serum for potential biomarker discovery in pancreatic cancer
Isomer-specific profiling of N-glycans derived from human serum for potential biomarker discovery in pancreatic cancer
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DOI：
10.1016/j.jprot.2018.04.016
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发表时间：
2018-06-15
影响因子：
3.3
通讯作者：
Liu, Xin
中科院分区：
文献类型：
--
作者：
Li, Yufei;Wang, Chang;Liu, Xin
糖基化是蛋白质翻译后最重要的修饰之一。近年来，人类血清糖组学的整体分析已经成为发现癌症相关生物标志物的非侵入性方法，并且许多研究都集中在组成性聚糖分析上。相比之下，结构特异性聚糖谱分析可以提供比组成谱分析具有更高特异性的更多潜在生物标志物。在这项工作中，从人血清中释放的N-聚糖用甲胺中和，并通过氨-硼烷复合物还原，然后使用具有多孔石墨化碳（PGC）的nanoLC-ESI-MS进行分析，并在胰腺癌（PC）病例（n = 32）和健康对照（n = 32）之间比较了超过280种异构体的相对丰度。统计分析鉴定了25种具有显著差异的特异性异构生物标志物（p值< 0.05）。进行ROC和PCA分析以评估被鉴定为在癌症中显著改变的潜在生物标志物。显著变化的特定异构体的AUC范围为0.712 - 0.949。此外，在所有潜在生物标志物的组合下，获得了更高的AUC 0.976，灵敏度（93.5%）和特异性（90.6%）。总的来说，所提出的策略加上异构聚糖的相对定量分析，使得有可能发现新的生物标志物用于诊断PC.Significance：胰腺癌（PC）的预后不良，5年生存率<5%。因此，确实需要准确诊断PC的策略。在本文中，一个双衍生化的策略，结构特异性聚糖分析已被使用，并根据我们最好的知识，这是第一次应用这种策略的PC生物标志物发现，其中异构体聚糖的分离，鉴定和相对定量可以同时获得。此外，通过对异构体聚糖的深入分析，还可以实现对聚糖立体和区域多样性的全面描述，这可能为发现PC生物标志物提供更多潜在信息。
Glycosylation is one of the most important post-translational modifications of protein. Recently, global profiling of human serum glycomics has become a noninvasive method for cancer-related biomarker discovery and many studies have focused on compositional glycan profiling. In contrast, structure-specific glycan profiling may provide more potential biomarkers with higher specificity than compositional profiling. In this work, N-glycans released from human serum were neutralized with methylamine and reduced by ammonia-borane complex prior to profiling using nanoLC-ESI-MS with porous graphitized carbon (PGC) and relative abundances of over 280 isomers were compared between pancreatic cancer (PC) cases (n = 32) and healthy controls (n = 32). Statistical analysis identified 25 specific-isomeric biomarkers with significant differences (p-value < 0.05). ROC and PCA analysis were performed to assess the potential biomarkers which were identified as being significantly altered in cancer. The AUCs of the significantly changed specific-isomers were ranging from 0.712 to 0.949. In addition, with the combination of all potential biomarkers, a higher AUC of 0.976 with sensitivity (93.5%) and specificity (90.6%) was obtained. Overall, the proposed strategy coupled to relative quantitative analysis of isomeric glycans make it possible to discover new biomarkers for the diagnosis of PC.Significance: Pancreatic cancer (PC) has a poor prognosis with a five-year survival rate < 5%. Therefore, a strategy for accurate diagnosis of PC is indeed required. In this paper, a dual-derivatized strategy for structure-specific glycan profiling has been used and according to our best knowledge, this is the first application of this strategy for PC biomarker discovery, in which the separation, identification and relative quantification of isomeric glycans can be simultaneously obtained. In addition, by in-depth analysis of isomeric glycans, the full description of the stereo- and region- diversity of glycans can also be achieved, which might provide more potential information for PC biomarker discovery.