Center for biomedical Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163, Mannheim, Germany.
HBIGS International Graduate School of Molecular and Cellular Biology, Heidelberg University, Im Neuenheimer Feld 501, 69120, Heidelberg, Germany.
Sci Rep. 2018 Jul 26;8(1):11260. doi: 10.1038/s41598-018-29677-z.
Recent advances in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry have enabled whole cell-MALDI mass spectrometry biotyping of drug-treated cultured cells for rapid monitoring of known abundant pharmacodynamic protein markers such as polyacetylated histones. In contrast, generic and automated analytical workflows for discovery of such pharmacodynamic markers, in particular lipid markers, and their use in cellular tests of drug-like compounds are still lacking. Here, we introduce such a workflow and demonstrate its utility for cellular drug-response monitoring of BCR-ABL tyrosine kinase inhibitors in K562 leukemia cells: First, low-molecular mass features indicating drug responses are computationally extracted from groups of MALDI-TOF mass spectra. Then, the lipids/metabolites corresponding to these features are identified by MALDI-Fourier transformation mass spectrometry. To demonstrate utility of the method, we identify the potassium adduct of phosphatidylcholine PC(36:1) as well as heme B, a marker for erythroid differentiation, as markers for a label-free MALDI MS-based test of cellular responses to BCR-ABL inhibitors. Taken together, these results suggest that MALDI-TOF mass spectrometry of lipids and other low molecular mass metabolites could support cell-based drug profiling.
基质辅助激光解吸/电离（MALDI）质谱技术的最新进展使得整个细胞 MALDI 质谱生物分型成为可能，可用于快速监测已知的丰富药效蛋白标志物，如多乙酰化组蛋白。相比之下，通用且自动化的分析工作流程用于发现此类药效标志物（特别是脂质标志物），并将其用于药物样化合物的细胞测试仍然缺乏。在这里，我们介绍了这样一种工作流程，并展示了其在 K562 白血病细胞中用于 BCR-ABL 酪氨酸激酶抑制剂的细胞药物反应监测的实用性：首先，从 MALDI-TOF 质谱的组中计算提取出指示药物反应的低分子量特征。然后，通过 MALDI-Fourier 变换质谱鉴定与这些特征相对应的脂质/代谢物。为了证明该方法的实用性，我们确定了 PC（36:1）磷脂的钾加合物以及血红素 B，一种红细胞分化的标志物，作为基于 MALDI MS 的无标记细胞对 BCR-ABL 抑制剂反应测试的标志物。总之，这些结果表明，脂质和其他低分子量代谢物的 MALDI-TOF 质谱可以支持基于细胞的药物分析。
