Triomics Analysis of Imatinib-Treated Myeloma Cells Connects Kinase Inhibition to RNA Processing and Decreased Lipid Biosynthesis.
Triomics Analysis of Imatinib-Treated Myeloma Cells Connects Kinase Inhibition to RNA Processing and Decreased Lipid Biosynthesis.
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DOI：
10.1021/acs.analchem.5b03040
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发表时间：
2015-11-03
影响因子：
7.4
通讯作者：
Asara JM
中科院分区：
文献类型：
--
作者：
Breitkopf SB;Yuan M;Helenius KP;Lyssiotis CA;Asara JM
在bcr - abl阳性的H929多发性骨髓瘤细胞上证实了代谢组学、脂质组学和磷蛋白质组学的结合，其中包括细胞培养中氨基酸（SILAC）蛋白质标记的三重稳定同位素标记，以及13C体内代谢物标记。从11880个磷酸化位点，我们证实H929细胞主要通过BCR-ABL-ERK途径信号传导，我们发现伊马替尼治疗不仅下调该途径中的磷酸化位点，还上调参与RNA表达的蛋白质上的磷酸化位点。代谢组学分析显示，H929细胞中的BCR-ABL-ERK信号通路驱动戊糖磷酸途径（PPP）和RNA生物合成，其中通过伊马替尼抑制该途径导致显著的PPP损伤、RNA核苷酸积累和mRNA的负调控。脂质组学数据还显示，脂质生物合成和脂肪酸掺入总体减少，溶血磷脂显著减少。RNA免疫沉淀研究证实，短期伊马替尼治疗抑制RNA降解，长期伊马替尼治疗抑制转录，验证了三组学的结果。这些数据显示了结合基于质谱的“组学”技术的实用性，并揭示了激酶抑制剂不仅可以下调其靶点的磷酸化，还可以通过增加其他细胞成分的磷酸化来诱导代谢事件。
The combination of metabolomics, lipidomics, and phosphoproteomics that incorporates triple stable isotope labeling by amino acids in cell culture (SILAC) protein labeling, as well as 13C in vivo metabolite labeling, was demonstrated on BCR–ABL-positive H929 multiple myeloma cells. From 11,880 phosphorylation sites, we confirm that H929 cells are primarily signaling through the BCR–ABL–ERK pathway, and we show that imatinib treatment not only downregulates phosphosites in this pathway but also upregulates phosphosites on proteins involved in RNA expression. Metabolomics analyses reveal that BCR–ABL–ERK signaling in H929 cells drives the pentose phosphate pathway (PPP) and RNA biosynthesis, where pathway inhibition via imatinib results in marked PPP impairment and an accumulation of RNA nucleotides and negative regulation of mRNA. Lipidomics data also show an overall reduction in lipid biosynthesis and fatty acid incorporation with a significant decrease in lysophospholipids. RNA immunoprecipitation studies confirm that RNA degradation is inhibited with short imatinib treatment and transcription is inhibited upon long imatinib treatment, validating the triomics results. These data show the utility of combining mass spectrometry-based “-omics” technologies and reveals that kinase inhibitors may not only downregulate phosphorylation of their targets but also induce metabolic events via increased phosphorylation of other cellular components.
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影响因子：
3.7
作者：
Caesar R;Manieri M;Kelder T;Boekschoten M;Evelo C;Müller M;Kooistra T;Cinti S;Kleemann R;Drevon CA
通讯作者：
Drevon CA
影响因子：
4.4
作者：
Christofk, Heather R.;Wu, Ning;Cantley, Lewis C.;Asara, John M.
通讯作者：
Asara, John M.
影响因子：
13.1
作者：
Cajka, Tomas;Fiehn, Oliver
通讯作者：
Fiehn, Oliver
影响因子：
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作者：
Buschow, Sonja I.;Lasonder, Edwin;Cavalieri, Duccio
通讯作者：
Cavalieri, Duccio
影响因子：
4.8
作者：
Choudhury, AD;Xu, H;Baer, R
通讯作者：
Baer, R