Cytometry by time-of-flight (CyTOF) simultaneously measures multiple cellular proteins at the single-cell level and is used to assess inter- and intra-tumor heterogeneity. This approach may be used to investigate the variability of individual tumor responses to treatments. Herein, we stratified lung tumor subpopulations based on AXL signaling as a potential targeting strategy. Integrative transcriptome analyses were used to investigate how TP-0903, an AXL kinase inhibitor, influences redundant oncogenic pathways in metastatic lung cancer cells. CyTOF profiling revealed that AXL inhibition suppressed SMAD4/TGF-β signaling and induced JAK1-STAT3 signaling to compensate for the loss of AXL. Interestingly, high JAK1-STAT3 was associated with increased levels of AXL in treatment-naïve tumors. Tumors with high AXL, TGF-β and JAK1 signaling concomitantly displayed CD133-mediated cancer stemness and hybrid EMT features in advanced stage patients, suggesting greater potential for distant dissemination. Diffusion pseudotime analysis revealed cell-fate trajectories among four different categories that were linked to clinicopathologic features for each patient. Patient-derived organoids (PDOs) obtained from tumors with high AXL and JAK1 were sensitive to TP-0903 and ruxolitinib (JAK inhibitor) treatments supporting the CyTOF findings. This study shows that single-cell proteomic profiling of treatment-naïve lung tumors, coupled with ex vivo testing of PDOs, identifies continuous AXL, TGF-β and JAK1-STAT3 signal activation in select tumors that may be targeted by combined AXL-JAK1 inhibition.

中文翻译：

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单细胞蛋白质组学分析确定联合 AXL 和 JAK1 抑制是肺癌的新治疗策略。

飞行时间流式细胞术 (CyTOF) 同时测量单细胞水平的多种细胞蛋白，用于评估肿瘤间和肿瘤内的异质性。这种方法可用于研究个体肿瘤对治疗反应的变异性。在此，我们基于 AXL 信号作为潜在的靶向策略对肺肿瘤亚群进行了分层。整合转录组分析用于研究 TP-0903（一种 AXL 激酶抑制剂）如何影响转移性肺癌细胞中的冗余致癌途径。CyTOF 分析显示 AXL 抑制抑制 SMAD4/TGF-β 信号传导并诱导 JAK1-STAT3 信号传导以补偿 AXL 的损失。有趣的是，高 JAK1-STAT3 与未经治疗的肿瘤中 AXL 水平升高有关。具有高 AXL 的肿瘤，TGF-β 和 JAK1 信号在晚期患者中同时显示出 CD133 介导的癌症干细胞和混合 EMT 特征，表明远处传播的潜力更大。扩散伪时间分析揭示了与每位患者的临床病理特征相关的四个不同类别之间的细胞命运轨迹。从具有高 AXL 和 JAK1 的肿瘤中获得的患者衍生的类器官 (PDO) 对支持 CyTOF 发现的 TP-0903 和鲁索替尼（JAK 抑制剂）治疗敏感。这项研究表明，对初治肺肿瘤的单细胞蛋白质组学分析，再加上 PDO 的离体测试，可以在可能通过联合 AXL-JAK1 抑制靶向的选定肿瘤中识别出持续的 AXL、TGF-β 和 JAK1-STAT3 信号激活.