Hu Youfan, Tang Jingyi, Sun Haoyi, Li Yanping, Yu Fengling, Zhang Guochen, Chen Jilan, Xu Huan, Zhong Zhanqiong, Huang Cong, Hu Kaifeng, Xie Mao, Yang Jiahui
School of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, 611137, China.
Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan 611137, China.
Phytomedicine. 2025 Oct;146:157085. doi: 10.1016/j.phymed.2025.157085. Epub 2025 Jul 21.
In non-small cell lung cancer (NSCLC), M2-polarized tumor-associated macrophages (TAMs) induce T cell dysfunction and correlate with poor clinical outcomes. Recent studies have demonstrated that artesunate (AS) enhances antitumor immunity, but the underlying mechanisms of AS reprogramming TAMs and boosting T cell anti-tumor activity in NSCLC need further clarification.
This study aims to clarify the molecular mechanisms by which AS induces TAM repolarization and enhances T cell activity to remodel the tumor microenvironment (TME) in NSCLC.
Lewis lung carcinoma (LLC)-bearing mice were treated with AS, and the subsets and activation status of immune cells in tumor were extensively analyzed. Subsequently, the effects of AS on macrophage polarization and T cell activation were evaluated in vitro which is further validated with publicly available clinical data. Finally, the therapeutic efficacy of AS combined with anti-PD-1 antibody was assessed in vivo.
In vivo studies demonstrated that AS inhibited LLC progression, enhanced T cell infiltration and activity, and reprogrammed TAMs into an inflammatory phenotype, thereby alleviating T cell exhaustion within the TME. RNA-Seq data from CD45 cells isolated from LLC tumors revealed that AS significantly upregulated genes involved in phagosome formation and antigen processing and presentation, with the interferon signaling emerging as one of the potential key pathways mediating these effects. In vitro experiments further confirmed that AS promoted macrophage reprogramming, enhancing their phagocytic activity, antigen cross-presentation, and tumoricidal capacity. Additionally, AS activated T cells directly or indirectly via STAT1/IRF1-driven macrophage repolarization. Finally, AS potentiated the tumoricidal efficacy of anti-PD-1 antibodies in the LLC mouse model.
Our study demonstrates that AS reprograms the TME through STAT1/IRF1-mediated M1 TAMs repolarization and T cell activation, broadening our understanding of the immunotherapeutic potential of AS and warranting further preclinical and clinical evaluations on combination of AS and PD-1 blockade for NSCLC treatment.
在非小细胞肺癌（NSCLC）中，M2极化的肿瘤相关巨噬细胞（TAM）诱导T细胞功能障碍，并与不良临床结果相关。最近的研究表明，青蒿琥酯（AS）可增强抗肿瘤免疫力，但AS在NSCLC中重新编程TAM和增强T细胞抗肿瘤活性的潜在机制需要进一步阐明。
本研究旨在阐明AS诱导TAM重极化并增强T细胞活性以重塑NSCLC肿瘤微环境（TME）的分子机制。
对携带Lewis肺癌（LLC）的小鼠进行AS治疗，并广泛分析肿瘤中免疫细胞的亚群和激活状态。随后，在体外评估AS对巨噬细胞极化和T细胞激活的影响，并用公开的临床数据进一步验证。最后，在体内评估AS联合抗PD-1抗体的治疗效果。
体内研究表明，AS抑制LLC进展，增强T细胞浸润和活性，并将TAM重新编程为炎症表型，从而减轻TME内的T细胞耗竭。从LLC肿瘤中分离的CD45细胞的RNA测序数据显示，AS显著上调了参与吞噬体形成以及抗原加工和呈递的基因，干扰素信号通路成为介导这些作用的潜在关键途径之一。体外实验进一步证实，AS促进巨噬细胞重编程，增强其吞噬活性、抗原交叉呈递和杀瘤能力。此外，AS通过STAT1/IRF1驱动的巨噬细胞重极化直接或间接激活T细胞。最后，AS增强了抗PD-1抗体在LLC小鼠模型中的杀瘤效果。
我们的研究表明，AS通过STAT1/IRF1介导的M1 TAM重极化和T细胞激活对TME进行重新编程，拓宽了我们对AS免疫治疗潜力的理解，并值得对AS与PD-1阻断联合用于NSCLC治疗进行进一步的临床前和临床评估。
