奥西替尼是第三代表皮生长因子受体酪氨酸激酶抑制剂，被批准作为具有 EGFR 激活突变或 T790M 耐药突变的晚期非小细胞肺癌 (NSCLC) 患者的一线治疗。然而，由于获得性耐药，奥希替尼的疗效受到限制，这凸显了阐明耐药机制以促进改进治疗策略的必要性。在这里，我们筛选了奥希替尼耐药 NSCLC 细胞中编码蛋白激酶显着上调的基因，并确定 NUAK1 是奥希替尼耐药的关键调节因子。 NUAK1在奥希替尼耐药的NSCLC中高表达并促进奥希替尼耐药的出现。 NUAK1 的遗传或药理学阻断可恢复耐药 NSCLC 细胞在体外和体内对奥希替尼的敏感性。从机制上讲，NUAK1 直接与 NADK 丝氨酸 64 (S64) 相互作用并磷酸化，从而减轻奥希替尼诱导的活性氧 (ROS) 积累，并导致 NSCLC 中获得奥希替尼耐药。此外，虚拟药物筛选发现T21195是NADK-S64磷酸化的抑制剂，并且T21195与奥希替尼协同作用，通过诱导ROS积累来逆转获得性耐药。总的来说，这些发现强调了 NUAK1-NADK 轴在控制 NSCLC 奥希替尼耐药中的作用，并表明靶向该轴作为规避耐药性的策略的潜力。

Osimertinib, a third-generation epidermal growth factor receptor tyrosine kinase inhibitor, is approved as a first-line therapy in advanced non-small cell lung carcinoma (NSCLC) patients with EGFR-activating mutations or the T790M resistance mutation. However, the efficacy of osimertinib is limited due to acquired resistance, highlighting the need to elucidate resistance mechanisms to facilitate the development of improved treatment strategies. Here, we screened for significantly upregulated genes encoding protein kinases in osimertinib-resistant NSCLC cells and identified NUAK1 as a pivotal regulator of osimertinib resistance. NUAK1 was highly expressed in osimertinib-resistant NSCLC and promoted the emergence of osimertinib resistance. Genetic or pharmacological blockade of NUAK1 restored the sensitivity of resistant NSCLC cells to osimertinib in vitro and in vivo. Mechanistically, NUAK1 directly interacted with and phosphorylated NADK at serine 64 (S64), which mitigated osimertinib-induced accumulation of reactive oxygen species (ROS) and contributed to the acquisition of osimertinib resistance in NSCLC. Furthermore, virtual drug screening identified T21195 as an inhibitor of NADK-S64 phosphorylation, and T21195 synergized with osimertinib to reverse acquired resistance by inducing ROS accumulation. Collectively, these findings highlight the role of the NUAK1-NADK axis in governing osimertinib resistance in NSCLC and indicate the potential of targeting this axis as a strategy for circumventing resistance.