Dissecting signaling regulators driving AXL-mediated bypass resistance and associated phenotypes by phosphosite perturbations.
Dissecting signaling regulators driving AXL-mediated bypass resistance and associated phenotypes by phosphosite perturbations.
复制标题
剖析通过磷酸位点扰动驱动 AXL 介导的旁路抗性和相关表型的信号调节因子。
DOI：
10.1101/2023.10.20.563266
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Meyer,AaronS
中科院分区：
文献类型：
--
作者：
Creixell,Marc;Taylor,ScottD;Gerritsen,Jacqueline;Bae,SongYi;Jiang,Mingxuan;Augustin,Teresa;Loui,Michelle;Boixo,Carmen;Creixell,Pau;White,ForestM;Meyer,AaronS
受体酪氨酸激酶(RTK)靶向治疗通常是有效的，但总是受到耐药性的限制。获得性耐药的一个主要机制涉及“旁路”切换到由恢复增殖的非靶向RTK驱动的替代途径。Axl就是这样一种RTK，它的过度表达经常见于旁路耐药肿瘤，它促进了细胞存活和相关的恶性表型，如上皮向间充质(EMT)的转化和迁移。然而，引发这些反应的信号分子和途径仍然难以捉摸。为了探索这些协同效应，我们产生了一组突变的肺腺癌PC9细胞系，在这些细胞系中，每个Axl细胞内的酪氨酸残基都突变为苯丙氨酸。通过多变量建模整合磷酸化信号和其他与抗性相关的表型变化的测量，我们将信号扰动映射到特定的抗性表型。我们的结果表明，Axl信号可以概括为与肺癌患者的进展性疾病和不良临床结局相关的两个簇。这些簇显示出良好的ABL1和SFK基序，并且它们的磷酸化被达沙替尼持续降低。高通量的激酶特异性分析表明，Ax1可能通过与Src形成复合体的FAK1激活SFK簇。此外，在肺癌细胞中，SFK簇与先前建立的与EMT介导的厄洛替尼耐药相关的粘着斑激酶(FAK1)信号重叠。最后，我们发现，在该激酶信号的下游，Ax1和Yap形成了一个正反馈环，支持耐药的持久细胞。总之，这项工作展示了一种剖析信号调节因子的方法，Ax1通过该方法驱动与erlotinib耐药相关的表型变化。
Receptor tyrosine kinase (RTK)-targeted therapies are often effective but invariably limited by drug resistance. A major mechanism of acquired resistance involves “bypass” switching to alternative pathways driven by non-targeted RTKs that restore proliferation. One such RTK is AXL whose overexpression, frequently observed in bypass resistant tumors, drives both cell survival and associated malignant phenotypes such as epithelial-to-mesenchymal (EMT) transition and migration. However, the signaling molecules and pathways eliciting these responses have remained elusive. To explore these coordinated effects, we generated a panel of mutant lung adenocarcinoma PC9 cell lines in which each AXL intracellular tyrosine residue was mutated to phenylalanine. By integrating measurements of phosphorylation signaling and other phenotypic changes associated with resistance through multivariate modeling, we mapped signaling perturbations to specific resistant phenotypes. Our results suggest that AXL signaling can be summarized into two clusters associated with progressive disease and poor clinical outcomes in lung cancer patients. These clusters displayed favorable Abl1 and SFK motifs and their phosphorylation was consistently decreased by dasatinib. High-throughput kinase specificity profiling showed that AXL likely activates the SFK cluster through FAK1 which is known to complex with Src. Moreover, the SFK cluster overlapped with a previously established focal adhesion kinase (FAK1) signature conferring EMT-mediated erlotinib resistance in lung cancer cells. Finally, we show that downstream of this kinase signaling, AXL and YAP form a positive feedback loop that sustains drug tolerant persister cells. Altogether, this work demonstrates an approach for dissecting signaling regulators by which AXL drives erlotinib resistance-associated phenotypic changes.