Phosphoproteomic Analysis of Leukemia Cells under Basal and Drug-treated Conditions Identifies Markers of Kinase Pathway Activation and Mechanisms of Resistance
Phosphoproteomic Analysis of Leukemia Cells under Basal and Drug-treated Conditions Identifies Markers of Kinase Pathway Activation and Mechanisms of Resistance
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DOI：
10.1074/mcp.m112.017483
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发表时间：
2012-08-01
影响因子：
7
通讯作者：
Cutillas, Pedro R.
中科院分区：
文献类型：
--
作者：
Alcolea, Maria P.;Casado, Pedro;Cutillas, Pedro R.
蛋白激酶信号是细胞稳态的基础，在所有癌症中都是失调的，但在患者之间有所不同。了解这种异质性的机制对于个性化靶向治疗至关重要。在这里，我们使用最近建立的LC-MS/MS平台来分析对激酶抑制剂具有不同敏感性的急性髓性白血病细胞系中的蛋白磷酸化。本研究中使用的化合物最初开发用于靶向Janus激酶、磷脂酰肌醇3-激酶和MEK。经过进一步验证的技术，我们确定了几个磷酸化位点，这些化合物的抑制，但其强度并不总是与生长抑制敏感性。相比之下，与敏感性/抗性相关的几百个磷酸化位点通常不被化合物抑制。这些结果表明，通路活性的标志物可能并不总是癌细胞对靶向这些通路的抑制剂的敏感性的可靠指标，因为平行激酶的活性可能有助于耐药性。通过挖掘我们的数据，我们确定了蛋白激酶C亚型作为在耐药细胞中更活跃的平行途径之一。与几种平行激酶途径导致耐药的观点一致，靶向蛋白激酶C、MEK和Janus激酶的抑制剂在阻止多药耐药细胞增殖方面相互增强。非靶向/无偏倚的方法，如本文所述，与平行激酶途径的活性一起定量预期靶激酶途径的活性，对于基于激酶抑制剂的个性化治疗将是非常宝贵的。Molecular & Cellular Proteomics 11：10.1074/mcp. M112.017483，453-466，2012.
Protein kinase signaling is fundamental to cell homeostasis and is deregulated in all cancers but varies between patients. Understanding the mechanisms underlying this heterogeneity is critical for personalized targeted therapies. Here, we used a recently established LC-MS/MS platform to profile protein phosphorylation in acute myeloid leukemia cell lines with different sensitivities to kinase inhibitors. The compounds used in this study were originally developed to target Janus kinase, phosphatidylinositol 3-kinase, and MEK. After further validation of the technique, we identified several phosphorylation sites that were inhibited by these compounds but whose intensities did not always correlate with growth inhibition sensitivity. In contrast, several hundred phosphorylation sites that correlated with sensitivity/ resistance were not in general inhibited by the compounds. These results indicate that markers of pathway activity may not always be reliable indicators of sensitivity of cancer cells to inhibitors that target such pathways, because the activity of parallel kinases can contribute to resistance. By mining our data we identified protein kinase C isoforms as one of such parallel pathways being more active in resistant cells. Consistent with the view that several parallel kinase pathways were contributing to resistance, inhibitors that target protein kinase C, MEK, and Janus kinase potentiated each other in arresting the proliferation of multidrug-resistant cells. Untargeted/unbiased approaches, such as the one described here, to quantify the activity of the intended target kinase pathway in concert with the activities of parallel kinase pathways will be invaluable to personalize therapies based on kinase inhibitors. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.017483, 453-466, 2012.