Zhou Shaoxuan, Wang Yueqin, Wei Jingyao, An Ke, Shi Yong, Zhang Xuran, Wang Han, Feng Luyao, He Lulu, Zhang Yu, Ren Tong, Li Ouwen, Yang Yun-Gui, Tian Xin
Department of Pharmacy, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Henan Key Laboratory of Precision Clinical Pharmacy, Zhengzhou University, Zhengzhou 450052, China.
Genomics Proteomics Bioinformatics. 2025 Aug 26. doi: 10.1093/gpbjnl/qzaf076.
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have produced durable complete responses, but the eventual development of acquired resistance presents a major challenge in the treatment of non-small cell lung cancer (NSCLC). N7-methylguanosine (m7G), a prevalent post-transcriptional modification within RNA, plays regulatory roles in RNA stability, expression dynamics, and functional diversity. Despite these insights, the contribution of m7G methylation to EGFR-TKIs resistance remains poorly characterized. Here, we demonstrate that internal m7G modifications of mRNA and their associated methyltransferase complex, methyltransferase-like 1 (METTL1)/WD repeat domain 4 (WDR4), are significantly elevated in NSCLC specimens, which correlates with therapeutic resistance. Functional assays confirmed that METTL1/WDR4 enhances gefitinib resistance in both cellular and animal models through internal RNA m7G methyltransferase activity in NSCLC. Mechanistically, m7G MeRIP-seq combined with RNA-seq identified sodium channel and clathrin linker 1 (SCLT1) as the m7G target of METTL1/WDR4. METTL1/WDR4 knockdown led to decreased methylation level and mRNA stability of the SCLT1 transcript. Importantly, overexpression of wild-type METTL1, but not its catalytically inactive mutant, restored mRNA stability. Furthermore, METTL1/WDR4-mediated m7G modification of SCLT1 regulates gefitinib resistance by activating the NF-κB signaling. Our findings reveal the crucial role of aberrant mRNA internal m7G modification in EGFR-TKIs resistance, suggesting that targeting the METTL1/WDR4-SCLT1-NF-κB axis holds a promising therapeutic potential for overcoming EGFR-TKIs resistance.
表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs）已产生持久的完全缓解，但获得性耐药的最终出现对非小细胞肺癌（NSCLC）的治疗构成了重大挑战。N7-甲基鸟苷（m7G）是RNA中一种普遍的转录后修饰，在RNA稳定性、表达动态和功能多样性中发挥调节作用。尽管有这些见解，但m7G甲基化对EGFR-TKIs耐药性的贡献仍未得到充分表征。在这里，我们证明mRNA的内部m7G修饰及其相关的甲基转移酶复合物，即甲基转移酶样1（METTL1）/WD重复结构域4（WDR4），在NSCLC标本中显著升高，这与治疗耐药性相关。功能试验证实，METTL1/WDR4通过NSCLC中的内部RNA m7G甲基转移酶活性增强细胞和动物模型中的吉非替尼耐药性。从机制上讲，m7G MeRIP-seq与RNA-seq相结合确定钠通道和网格蛋白连接蛋白1（SCLT）为METTL1/WDR4的m7G靶点。METTL1/WDR4敲低导致SCLT1转录本的甲基化水平和mRNA稳定性降低。重要的是，野生型METTL1的过表达，而不是其催化失活的突变体，恢复了mRNA稳定性。此外，METTL1/WDR4介导的SCLT1的m7G修饰通过激活NF-κB信号传导来调节吉非替尼耐药性。我们的研究结果揭示了异常的mRNA内部m7G修饰在EGFR-TKIs耐药性中的关键作用，表明靶向METTL1/WDR4-SCLT1-NF-κB轴具有克服EGFR-TKIs耐药性的潜在治疗前景。
