Epidermal growth factor receptor (EGFR) is one of the most common amplified and overexpressed oncogenes in esophageal squamous cell carcinoma (ESCC), while the clinical efficacy of EGFR-targeted therapy in ESCC is dismal. Here, we evaluated the efficacy of dual blockage using monoclonal antibody against EGFR (Nimotuzumab) and an Wee1 inhibitor (AZD1775) in ESCC. We found that the mRNA and protein expression of EGFR and Wee1 were positively correlated in ESCC. Nimotuzumab-AZD1775 co-treatment inhibited tumor growth in PDX models with different drug susceptibility. Transcriptome sequencing and mass spectrometry analysis indicated that higher sensitive models showed enrichment of the PI3K/Akt or MAPK signaling pathway in Nimotuzumab-AZD1775 group compared with control group. In vitro experiments showed that the combination further inhibit PI3K/Akt and MAPK pathways compared to their monotherapy as indicated by downregulation of pAKT, pS6, pMEK, pErk and p-p38 MAPK. Furthermore, AZD1775 potentiated Nimotuzumab's antitumor effect through inducing apoptosis. Meanwhile, the bioinformatics analysis suggests the POLR2A might be candidate molecule of EGFR/Wee1 downstream. In conclusion, our work uncovers that EGFR-mAb Nimotuzumab combined with Wee1 inhibitor AZD1775 elicited potentiated anticancer activity against ESCC cell line and PDXs partially through PI3K/Akt and MAPK pathways blockade. These preclinical data raise the promising that ESCC patients may benefit from dual target EGFR and Wee1.

中文翻译：

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Wee1 和 EGFR 联合抑制揭示了食管鳞状细胞癌的协同抗肿瘤作用。

表皮生长因子受体（EGFR）是食管鳞状细胞癌（ESCC）中最常见的扩增和过度表达的癌基因之一，而EGFR靶向治疗食管鳞状细胞癌的临床疗效却很差。在这里，我们评估了使用抗 EGFR 单克隆抗体（尼妥珠单抗）和 Wee1 抑制剂 (AZD1775) 在 ESCC 中的双重阻断效果。我们发现ESCC中EGFR和Wee1的mRNA和蛋白表达呈正相关。Nimotuzumab-AZD1775 联合治疗可抑制不同药敏 PDX 模型中的肿瘤生长。转录组测序和质谱分析表明，与对照组相比，更高灵敏度的模型显示 Nimotuzumab-AZD1775 组的 PI3K/Akt 或 MAPK 信号通路富集。体外实验表明，与单一疗法相比，该组合进一步抑制 PI3K/Akt 和 MAPK 通路，pAKT、pS6、pMEK、pErk 和 p-p38 MAPK 下调表明该组合。此外，AZD1775 通过诱导细胞凋亡增强 Nimotuzumab 的抗肿瘤作用。同时，生物信息学分析表明POLR2A可能是EGFR/Wee1下游的候选分子。总之，我们的工作发现，EGFR-mAb Nimotuzumab 与 Wee1 抑制剂 AZD1775 联合可部分通过 PI3K/Akt 和 MAPK 通路阻断来增强针对 ESCC 细胞系和 PDX 的抗癌活性。这些临床前数据表明 ESCC 患者可能受益于双靶点 EGFR 和 Wee1。