Jie Hui, Lai Hongjin, Wang Zihuai, Yi Min, Liu Yi, Urbanska Edyta Maria, Santoni-Rugiu Eric, Wei Shiyou, Chen Yuhao, Li Chuan, Wang Tengyong, Luo Nanzhi, Liu Lunxu, Deng Senyi, Guo Chenglin
Department of Thoracic Surgery and Institute of Thoracic Oncology, West China Hospital, Sichuan University, Chengdu, China.
Department of Oncology, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark.
Transl Lung Cancer Res. 2025 Mar 31;14(3):878-896. doi: 10.21037/tlcr-2025-105. Epub 2025 Mar 27.
As an approved targeting drug, crizotinib has been widely used in the treatment of patients with non-small cell lung cancer (NSCLC) with anaplastic lymphoma kinase () rearrangements or c-ros oncogene 1 () fusions and has demonstrated remarkable therapeutic effects. However, crizotinib-treated patients frequently experience drug resistance, and there are still some underlying mechanisms, which remain unclear. Autophagy, a cellular process that involves the degradation and recycling of cellular components, has been implicated in the development of drug resistance. In this study, we aim to elucidate the mechanisms of crizotinib resistance involving autophagy dysregulation and identify novel therapeutic targets to overcome this resistance.
We first established a model for crizotinib resistance in HCC78 and H3122 cells. Next, the level of proliferation, apoptosis, autophagy flux, and reactive oxygen species (ROS) of these cells were measured. Subsequently, we analyzed the published single-cell RNA sequencing data from three -rearranged lung cancer organoid samples and performed a metabolomics assay on crizotinib-resistant HCC78 cells. Finally, the therapeutic effects were confirmed by targeting autophagy flux.
Crizotinib induced cell apoptosis and growth arrest by promoting the accumulation of autophagosomes through the inhibition of autophagy flux in or NSCLC. In contrast, crizotinib-resistant NSCLC cells showed inactivation of signal transducer and activator of transcription 3 (STAT3) phosphorylation and downregulation of prostaglandin endoperoxide synthase 2 (), leading to an increase in the metabolite arachidonic acid (AA). AA further promoted autophagy flux and reduced autophagosome accumulation, driving crizotinib resistance under conditions of drug stress. Moreover, chloroquine (CQ), anti-malaria drug and lysosome inhibitor developed in 1940, could induce cell death in crizotinib-resistant NSCLC by blocking AA-mediated autophagy flux and facilitating autophagosome accumulation, significantly enhancing the treatment efficacy of crizotinib in drug-resistant NSCLC.
We discovered a new mechanism of first generation ALK- and ROS1-TKIs resistance, which points to the role of the metabolite AA in resistance to tyrosine kinase inhibitors. It may potentially provide an alternative strategy to overcoming crizotinib resistance in NSCLC treatment by reversing AA-mediated autophagy.
克唑替尼作为一种获批的靶向药物，已广泛用于治疗间变性淋巴瘤激酶（ALK）重排或c-ros原癌基因1（ROS1）融合的非小细胞肺癌（NSCLC）患者，并显示出显著的治疗效果。然而，接受克唑替尼治疗的患者经常出现耐药性，其潜在机制仍不明确。自噬是一种涉及细胞成分降解和循环利用的细胞过程，与耐药性的产生有关。在本研究中，我们旨在阐明克唑替尼耐药涉及自噬失调的机制，并确定克服这种耐药性的新治疗靶点。
我们首先在HCC78和H3122细胞中建立了克唑替尼耐药模型。接下来，检测这些细胞的增殖、凋亡、自噬通量和活性氧（ROS）水平。随后，我们分析了来自三个ALK重排肺癌类器官样本的已发表单细胞RNA测序数据，并对克唑替尼耐药的HCC78细胞进行了代谢组学分析。最后，通过靶向自噬通量来确认治疗效果。
克唑替尼通过抑制ALK或ROS1 NSCLC中的自噬通量，促进自噬体的积累，从而诱导细胞凋亡和生长停滞。相反，克唑替尼耐药的NSCLC细胞显示信号转导和转录激活因子3（STAT3）磷酸化失活以及前列腺素内过氧化物合酶2（COX2）下调，导致代谢物花生四烯酸（AA）增加。AA进一步促进自噬通量并减少自噬体积累，在药物应激条件下导致克唑替尼耐药。此外，1940年开发的抗疟疾药物和溶酶体抑制剂氯喹（CQ），可通过阻断AA介导的自噬通量并促进自噬体积累，诱导克唑替尼耐药的NSCLC细胞死亡，显著提高克唑替尼在耐药NSCLC中的治疗效果。
我们发现了第一代ALK和ROS1酪氨酸激酶抑制剂耐药的新机制，这表明代谢物AA在对酪氨酸激酶抑制剂耐药中的作用。它可能为通过逆转AA介导的自噬来克服NSCLC治疗中克唑替尼耐药提供一种替代策略。
