Rapid and comprehensive 'shotgun' lipidome profiling of colorectal cancer cell derived exosomes.
Rapid and comprehensive 'shotgun' lipidome profiling of colorectal cancer cell derived exosomes.
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DOI：
10.1016/j.ymeth.2015.04.014
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发表时间：
2015-10-01
期刊：
影响因子：
--
通讯作者：
Reid GE
中科院分区：
文献类型：
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作者：
Lydic TA;Townsend S;Adda CG;Collins C;Mathivanan S;Reid GE
越来越多的人认识到癌细胞来源的外切体在与靶细胞融合或摄取时在细胞间信号转导中所起的作用，包括免疫系统逃避、肿瘤生长和转移。然而，到目前为止，尽管外体膜和货脂被认为在外体的生物发生和分泌以及融合或摄取和靶细胞功能反应中发挥关键作用，但癌细胞来源的外体脂在一系列不同癌症中的详细特征尚未得到广泛的探索。在这里，一个简单和直接的脂体分析策略，包括优化的样品提取和新的样品衍生化技术，结合高分辨率的‘鸟枪式’质谱学和‘靶向’串联质谱学方法，被证明可以快速从结直肠癌细胞系LIM1215分泌的外切体中鉴定36个脂类和亚类中的>；520种单个脂类。Exosome与细胞脂质谱的相对定量和比较显示，某些脂类显著丰富，以及大量的脂亚类重塑和单个脂类的丰度变化，包括鞘脂、甾醇脂、甘油脂和甘油磷脂，特别是含血浆蛋白原和烷基醚的甘油磷脂。因此，这一分析策略为广泛的癌细胞或组织衍生的外切体提供了一个全面的脂体图谱平台，将有助于后续的功能研究，旨在阐明特定的细胞或外切体脂质在结直肠癌的发生和发展中的作用，或确定可作为有效诊断或预后疾病生物标志物的特定脂质(S)。
There is an increasing recognition of the role that cancer cell derived exosomes play in intercellular signaling upon fusion or uptake with a target cell, including immune system evasion, tumor growth and metastasis. To date, however, although exosomal membrane and cargo lipids are expected to play a pivotal role in exosome biogenesis and secretion, as well as in fusion or uptake and target cell functional response, the detailed characterization of cancer cell derived exosome lipids across a range of different cancers has not yet been broadly explored. Here, a simple and straightforward lipidome analysis strategy consisting of optimized sample extraction and novel sample derivatization techniques, coupled with high-resolution ‘shotgun’ mass spectrometry and ‘targeted’ tandem mass spectrometry methods, is demonstrated for the rapid identification of >520 individual lipids in 36 lipid classes and sub classes from exosomes secreted by the colorectal cancer cell line, LIM1215. Relative quantification and comparison of exosome versus cellular lipid profiles reveals significant enrichment of certain lipid classes, as well as substantial lipid subclass remodeling and changes in abundance of individual lipids, including sphingolipids, sterol lipids, glycerolipids and glycerophospholipids, and particularly plasmalogen- and alkyl ether-containing glycerophospholipids. This analysis strategy therefore provides a platform for comprehensive lipidome profiling across a wide range of cancer cell or tissue derived exosomes, that will facilitate subsequent functional studies aimed at elucidating the role of specific cellular or exosome lipids in the onset and progression of colorectal cancer, or to identify specific lipid(s) that could serve as effective diagnostic or prognostic disease biomarkers.