Integrated Proteomics-Based Physical and Functional Mapping of AXL Kinase Signaling Pathways and Inhibitors Define Its Role in Cell Migration.
Integrated Proteomics-Based Physical and Functional Mapping of AXL Kinase Signaling Pathways and Inhibitors Define Its Role in Cell Migration.
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DOI：
10.1158/1541-7786.mcr-21-0275
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发表时间：
2022-04-01
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影响因子：
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通讯作者：
Haura EB
中科院分区：
文献类型：
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作者：
Majumder A;Hosseinian S;Stroud M;Adhikari E;Saller JJ;Smith MA;Zhang G;Agarwal S;Creixell M;Meyer BS;Kinose F;Bowers K;Fang B;Stewart PA;Welsh EA;Boyle TA;Meyer AS;Koomen JM;Haura EB
为了更好地理解TAM受体酪氨酸激酶家族成员AXL的信号传导复杂性，我们创建了三种AXL酪氨酸激酶抑制剂（TKI）的AXL信号传导相互作用、磷酸化事件和靶点接合的物理和功能图。我们使用BioID评估了AXL蛋白复合物，使用质谱法评估了AXL TKI对全局磷蛋白的影响，并使用基于活性的蛋白质分析评估了AXL TKI的靶向结合。BioID鉴定了主要参与细胞粘附/迁移的AXL相互作用蛋白。全球磷酸化蛋白质组学显示，AXL抑制降低了参与磷脂酰肌醇介导的信号传导和细胞粘附/迁移的肽的磷酸化。三种AXL抑制剂的比较表明，TKI RXDX-106抑制pAXL、pAKT和这些细胞的迁移/侵袭，而不降低其活力，而Bemcentinib对活力发挥AXL非依赖性表型效应。这些TKI的蛋白质组学表征表明，它们抑制除AXL外的多种靶标，其中Bemcentinib具有最多的脱靶。AXL和EGFR TKI联合治疗未逆转厄洛替尼耐药细胞系模型中的耐药性。然而，在一个抗性克隆中鉴定出独特的脆弱性，其中Bemcentinib和厄洛替尼的组合抑制细胞活力和信号传导。我们还表明，AXL在约30-40%的非小细胞肺癌中过表达，但在小细胞肺癌中很少过表达。细胞系具有广泛的AXL表达，很少检测到基础激活。我们的研究定义了AXL在肺癌中的作用机制，可用于建立测定患者组织中药物靶向活性AXL复合物的测定方法，并为靶向其信号传导作为抗癌治疗提供信息。
To better understand the signaling complexity of AXL, a member of the TAM receptor tyrosine kinase family, we created a physical and functional map of AXL signaling interactions, phosphorylation events, and target-engagement of three AXL tyrosine kinase inhibitors (TKI). We assessed AXL protein-complexes using BioID, effects of AXL TKI on global phosphoproteins using mass spectrometry, and target engagement of AXL TKI using activity-based protein profiling. BioID identifies AXL-interacting proteins that are mostly involved in cell adhesion/migration. Global phosphoproteomics show that AXL inhibition decreases phosphorylation of peptides involved in phosphatidylinositol-mediated signaling and cell adhesion/migration. Comparison of three AXL inhibitors reveals that TKI RXDX-106 inhibits pAXL, pAKT and migration/invasion of these cells without reducing their viability, while Bemcentinib exerts AXL-independent phenotypic effects on viability. Proteomic characterization of these TKIs demonstrates that they inhibit diverse targets in addition to AXL, with Bemcentinib having the most off-targets. AXL and EGFR TKI co-treatment did not reverse resistance in cell line models of Erlotinib-resistance. However, a unique vulnerability was identified in one resistant clone, wherein combination of Bemcentinib and Erlotinib inhibited cell viability and signaling. We also show that AXL is overexpressed in ~30–40% of non-small but rarely in small-cell lung cancer. Cell lines have a wide range of AXL expression, with basal activation detected rarely. Our study defines mechanisms of action of AXL in lung cancers which can be used to establish assays to measure drug targetable active AXL-complexes in patient tissues and inform the strategy for targeting it’s signaling as an anticancer therapy.