The insertion site of the internal tandem duplications (ITDs) in the FLT3 gene affects the sensitivity to tyrosine kinase inhibitors (TKIs) therapy in acute myeloid leukemia (AML). Patients with the ITD in the tyrosine kinase domain lack effective therapeutic options. Here, to identify genotype-driven strategies increasing the TKI therapy efficacy, we developed SignalingProfiler, a strategy supporting the integration of high-sensitive mass spectrometry-based (phospho)proteomics, RNA sequencing datasets with literature-derived signaling networks. The approach generated FLT3-ITD genotype-specific predictive models and revealed a conserved role of the WEE1-CDK1 axis in TKIs resistance. Remarkably, pharmacological inhibition of the WEE1 kinase synergizes and strengthens the pro-apoptotic effect of TKIs therapy in cell lines and patient-derived primary blasts. Finally, we propose a new molecular mechanism of TKIs resistance in AML and suggest the combination of WEE1 inhibitor and TKI as a therapeutic option to improve patients clinical outcome.

FLT3 基因中内部串联重复 (ITD) 的插入位点影响急性髓系白血病 (AML) 对酪氨酸激酶抑制剂 (TKI) 治疗的敏感性。酪氨酸激酶结构域 ITD 患者缺乏有效的治疗选择。在这里，为了确定提高 TKI 治疗功效的基因型驱动策略，我们开发了SignalingProfiler，一种支持将基于高灵敏度质谱的（磷酸）蛋白质组学、RNA 测序数据集与文献衍生的信号网络整合的策略。该方法生成了 FLT3-ITD 基因型特异性预测模型，并揭示了 WEE1-CDK1 轴在 TKI 耐药中的保守作用。值得注意的是，WEE1 激酶的药理抑制可协同并增强 TKI 治疗在细胞系和患者来源的原代细胞中的促凋亡作用。最后，我们提出了 AML 中 TKI 耐药的新分子机制，并建议将 WEE1 抑制剂与 TKI 联合作为改善患者临床结果的治疗选择。