Lu Conghua, Yu Rui, Zhang Chong, Lin Caiyu, Dou Yuanyao, Wu Di, Pan Yonghong, Peng Tao, Tang Huan, Han Rui, He Yong
Department of Respiratory Disease, Daping Hospital, Army Medical University, 400042, Chongqing, China.
Department of Ultrasound, The First Affiliated Hospital of Chongqing Medical University, 400042, Chongqing, China.
Cell Death Discov. 2022 Apr 22;8(1):221. doi: 10.1038/s41420-022-01027-z.
Lorlatinib is a promising third-generation anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitor (TKI) that has been approved for treating ALK-positive non-small-cell lung cancer (NSCLC) patients with previous ALK-TKI treatment failures. However, the inevitable emergence of acquired resistance limits its long-term efficacy. A more comprehensive understanding of the acquired resistance mechanisms to lorlatinib will enable the development of more efficacious therapeutic strategies. The efficacy of chloroquine (CQ) in combination with lorlatinib in ALK-positive NSCLC cells in vitro and in vivo was assessed using CCK-8, colony formation, immunofluorescence staining, flow cytometry analysis, western blot analysis, and xenograft implantation. Here, we show that lorlatinib induced apoptosis and protective autophagy in ALK-positive NSCLC cells. However, the protective autophagy can gradually lead to decreased cytotoxicity of loratinib in ALK-positive NSCLC cells. Meanwhile, we found that the combination of lorlatinib and CQ, an inhibitor of autophagy, inhibited autophagy and promoted apoptosis both in vitro and in vivo, which sensitized cells to lorlatinib through the dephosphorylation of Foxo3a and promoted nuclear translocation, then activation of Foxo3a/Bim axis. Taken together, our results suggest that inhibition of protective autophagy might be a therapeutic target for delaying the occurrence of acquired resistance to lorlatinib in ALK-positive NSCLC patients.
劳拉替尼是一种很有前景的第三代间变性淋巴瘤激酶（ALK）酪氨酸激酶抑制剂（TKI），已被批准用于治疗既往接受ALK-TKI治疗失败的ALK阳性非小细胞肺癌（NSCLC）患者。然而，获得性耐药的不可避免出现限制了其长期疗效。更全面地了解劳拉替尼的获得性耐药机制将有助于开发更有效的治疗策略。使用CCK-8、集落形成、免疫荧光染色、流式细胞术分析、蛋白质免疫印迹分析和异种移植植入评估了氯喹（CQ）与劳拉替尼联合应用于体外和体内ALK阳性NSCLC细胞的疗效。在此，我们表明劳拉替尼在ALK阳性NSCLC细胞中诱导凋亡和保护性自噬。然而，保护性自噬可逐渐导致劳拉替尼在ALK阳性NSCLC细胞中的细胞毒性降低。同时，我们发现劳拉替尼与自噬抑制剂CQ联合应用在体外和体内均抑制自噬并促进凋亡，通过Foxo3a的去磷酸化使细胞对劳拉替尼敏感并促进核转位，进而激活Foxo3a/Bim轴。综上所述，我们的结果表明抑制保护性自噬可能是延缓ALK阳性NSCLC患者对劳拉替尼获得性耐药发生的治疗靶点。
