Department of Biochemistry, University of Zurich, Winterthurerstr. 190, 8057 Zurich, Switzerland.
Department of Biological Engineering, Koch Institute for Integrative Cancer Research, Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Sci Signal. 2019 Jan 22;12(565):eaau2875. doi: 10.1126/scisignal.aau2875.
Drug-induced compensatory signaling and subsequent rewiring of the signaling pathways that support cell proliferation and survival promote the development of acquired drug resistance in tumors. Here, we sought to analyze the adaptive kinase response in cancer cells after distinct treatment with agents targeting human epidermal growth factor receptor 2 (HER2), specifically those that induce either only temporary cell cycle arrest or, alternatively, apoptosis in HER2-overexpressing cancers. We compared trastuzumab, ARRY380, the combination thereof, and a biparatopic, HER2-targeted designed ankyrin repeat protein (DARPin; specifically, 6L1G) and quantified the phosphoproteome by isobaric tagging using tandem mass tag liquid chromatography/tandem mass spectrometry (TMT LC-MS/MS). We found a specific signature of persistently phosphorylated tyrosine peptides after the nonapoptotic treatments, which we used to distinguish between different treatment-induced cancer cell fates. Next, we analyzed the activation of serine/threonine and tyrosine kinases after treatment using a bait peptide chip array and predicted the corresponding active kinases. Through a combined system-wide analysis, we identified a common adaptive kinase response program that involved the activation of focal adhesion kinase 1 (FAK1), protein kinase C-δ (PRKCD), and Ephrin (EPH) family receptors. These findings reveal potential targets to prevent adaptive resistance to HER2-targeted therapies.
药物诱导的补偿性信号转导和随后支持细胞增殖和存活的信号通路的重新布线促进了肿瘤获得性耐药的发展。在这里，我们试图分析在针对人表皮生长因子受体 2（HER2）的药物（特别是那些仅诱导暂时性细胞周期停滞或HER2 过表达癌症中的细胞凋亡的药物）对癌细胞进行不同处理后适应性激酶反应。我们比较了曲妥珠单抗、 ARRY380、它们的组合以及双靶向、HER2 靶向设计的锚蛋白重复蛋白（DARPin；具体为 6L1G），并通过使用串联质量标签液相色谱/串联质谱（TMT LC-MS/MS）进行等压标记来定量磷酸蛋白质组。我们发现非凋亡处理后存在持续磷酸化酪氨酸肽的特定特征，我们将其用于区分不同治疗诱导的癌细胞命运。接下来，我们使用诱饵肽芯片阵列分析处理后丝氨酸/苏氨酸和酪氨酸激酶的激活，并预测相应的活性激酶。通过综合系统分析，我们确定了一个共同的适应性激酶反应程序，涉及粘着斑激酶 1（FAK1）、蛋白激酶 C-δ（PRKCD）和 Ephrin（EPH）家族受体的激活。这些发现揭示了预防 HER2 靶向治疗获得性耐药的潜在靶点。
