Wu Jie, Liu Nian, Chen Jing, Tao Qian, Lu Can, Li Qiuqiu, Chen Xiang, Peng Cong
Department of Dermatology, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Hunan Key Laboratory of Skin Cancer and Psoriasis, Human Engineering Research Center of Skin Health and Disease, Xiangya Hospital, Central South University, Changsha, Hunan, China.
J Immunother Cancer. 2025 Feb 6;13(2):e010252. doi: 10.1136/jitc-2024-010252.
Clofarabine (Clo) is a Food and Drug Administration (FDA)-approved drug for the treatment of acute lymphoblastic leukemia; however, its effects on solid tumors remain largely unknown.
In vitro and in vivo experiments have demonstrated the cytotoxic effects of Clo on melanoma and lung cancer. The molecular mechanisms of Clo-induced tumor cell death were analyzed using western blotting, ELISA, reverse transcription-PCR, immunofluorescence, co-immunoprecipitation (CO-IP), short hairpin RNA, co-culture, chromatin immunoprecipitation, and flow cytometry. Clinical data sets were used to analyze the correlation between stimulator of interferon genes (STING)-NFκB signaling and immune infiltration.
In this study, Clo significantly reduced the growth of melanoma and lung cancer cells. Furthermore, Clo treatment induced GSDME-mediated pyroptosis. Most importantly, Clo administration dramatically increased the cytotoxic activity of CD8 T cells in vitro and in vivo. Mechanistically, the administration of Clo induced the interaction of P53 with STING, which activated the non-canonical STING-NFκB pathway; consequently, NF-κB directly bound to the promoter regions of its target genes, including CCL5, CXCL10, HLAs and BAX. This resulted in apoptosis, pyroptosis, and immunogenic cell death in tumor cells by Clo. Furthermore, Clo-induced GSDME-mediated pyroptosis partly assists in activating T cell immunity via CCL5 and CXCL10. The non-canonical STING-NF-κB pathway is the crucial signaling pathway that initiates and links apoptosis, pyroptosis, and immunogenic cell death.
Our study is the first to show that Clo, an FDA-approved drug, induces tumor cell apoptosis, GSDME-related pyroptosis, and CD8 T-cell antitumor activity via the non-canonical P53-STING-NF-κB signaling pathway, providing a novel strategy for the clinical therapy of melanoma and lung cancer.
氯法拉滨（Clo）是一种经美国食品药品监督管理局（FDA）批准用于治疗急性淋巴细胞白血病的药物；然而，其对实体瘤的作用在很大程度上仍不清楚。
体外和体内实验已证明Clo对黑色素瘤和肺癌具有细胞毒性作用。使用蛋白质免疫印迹法、酶联免疫吸附测定、逆转录聚合酶链反应、免疫荧光、免疫共沉淀（CO-IP）、短发夹RNA、共培养、染色质免疫沉淀和流式细胞术分析Clo诱导肿瘤细胞死亡的分子机制。利用临床数据集分析干扰素基因刺激因子（STING）-核因子κB信号传导与免疫浸润之间的相关性。
在本研究中，Clo显著降低了黑色素瘤和肺癌细胞的生长。此外，Clo治疗诱导了GSDME介导的细胞焦亡。最重要的是，给予Clo在体外和体内均显著增加了CD8 T细胞的细胞毒性活性。从机制上讲，给予Clo诱导P53与STING相互作用，激活非经典的STING-核因子κB途径；因此，核因子κB直接结合其靶基因（包括CCL5、CXCL10、人类白细胞抗原和BAX）的启动子区域。这导致Clo诱导肿瘤细胞发生凋亡、细胞焦亡和免疫原性细胞死亡。此外，Clo诱导的GSDME介导的细胞焦亡部分通过CCL5和CXCL10协助激活T细胞免疫。非经典的STING-核因子κB途径是启动并连接凋亡、细胞焦亡和免疫原性细胞死亡的关键信号通路。
我们的研究首次表明，一种经FDA批准的药物Clo通过非经典的P53-STING-核因子κB信号通路诱导肿瘤细胞凋亡、GSDME相关的细胞焦亡以及CD8 T细胞抗肿瘤活性，为黑色素瘤和肺癌的临床治疗提供了一种新策略。
