LOBSTAHS: An Adduct-Based Lipidomics Strategy for Discovery and Identification of Oxidative Stress Biomarkers
LOBSTAHS: An Adduct-Based Lipidomics Strategy for Discovery and Identification of Oxidative Stress Biomarkers
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DOI：
10.1021/acs.analchem.6b01260
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发表时间：
2016-07-19
影响因子：
7.4
通讯作者：
Van Mooy, Benjamin A. S.
中科院分区：
文献类型：
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作者：
Collins, James R.;Edwards, Bethanie R.;Van Mooy, Benjamin A. S.
在大型LC/MS数据集中发现和鉴定分子生物标志物需要显著的自动化，而不会损失化合物筛选和注释过程的准确性。在这里，我们描述了一个脂质组学工作流程和开源软件包，用于高通量注释和推定识别高质量精度HPLC-MS数据中的脂质、氧化脂质和氧化脂质生物标志物。通过加合物层次序列或LOBSTAHS进行的脂质和氧脂平生物标志物筛选使用基于加合物离子形成模式和其他性质的正交筛选标准来识别数千种化合物，同时为用户提供每个分配的置信度得分。数据库由两个可自定义数据库之一组成;默认数据库包含14068个唯一条目。为了证明该软件的功能，我们筛选了超过340 000个质谱特征的实验中，过氧化氢被用来诱导海洋硅藻三角褐指藻的氧化应激。LOBSTAHS puppet在21869个特征中鉴定了1969个独特的母体化合物，这些化合物在多级筛选过程中幸存下来。虽然三角褐指藻在氧化应激下保持其核心脂质组的92%以上，但生物标志物分布和丰度的模式表明重塑是微妙和普遍的。用150 AcM H2 O2处理促进了统计学上显著的跨脂质类的碳链延长，最强的延长伴随着单半乳糖基甘油二酯部分的氧化，脂质通常定位于叶绿体。氧化应激也诱导脂质体峰面积显著重新分配至三酰甘油。LOBSTAHS可用于各种系统的环境或实验数据，并可在https://github.com/vanmooylipidomics/LOBSTAHS上免费获得。
Discovery and identification of molecular biomarkers in large LC/MS data sets requires significant automation without loss of accuracy in the compound screening and annotation process. Here, we describe a lipidomics workflow and open-source software package for high-throughput annotation and putative identification of lipid, oxidized lipid, and oxylipin biomarkers in high-mass-accuracy HPLC-MS data. Lipid and oxylipin biomarker screening through adduct hierarchy sequences, or LOBSTAHS, uses orthogonal screening criteria based on adduct ion formation patterns and other properties to identify thousands of compounds while providing the user with a confidence score for each assignment. Assignments are made from one of two customizable databases; the default databases contain 14 068 unique entries. To demonstrate the software's functionality, we screened more than 340 000 mass spectral features from an experiment in which hydrogen peroxide was used to induce oxidative stress in the marine diatom Phaeodactylum tricornutum. LOBSTAHS putatively identified 1969 unique parent compounds in 21 869 features that survived the multistage screening process. While P. tricornutum maintained more than 92% of its core lipidome under oxidative stress, patterns in biomarker distribution and abundance indicated remodeling was both subtle and pervasive. Treatment with 150 AcM H2O2 promoted statistically significant carbon-chain elongation across lipid classes, with the strongest elongation accompanying oxidation in moieties of monogalactosyldiacylglycerol, a lipid typically localized to the chloroplast. Oxidative stress also induced a pronounced reallocation of lipidome peak area to triacylglycerols. LOBSTAHS can be used with environmental or experimental data from a variety of systems and is freely available at https://github.com/vanmooylipidomics/LOBSTAHS.