Chen Ying, Li Qiaoming, Wang Zixiang, Sun Ling V, Hou Steven X
Department of Cell and Developmental Biology at School of Life Sciences, State Key Laboratory of Genetic Engineering, Institute of Metabolism and Integrative Biology, Human Phenome Institute, Children's Hospital, Zhongshan Hospital, Fudan University, Shanghai, 200438, China.
J Transl Med. 2025 May 20;23(1):561. doi: 10.1186/s12967-025-06576-2.
The immunosuppressive nature of the tumor microenvironment (TME) and the existence of cancer stem cells (CSCs) present significant hurdles in tumor therapy. The identification of therapeutic agents that can target both CSCs and the TME could be a potential approach to overcome treatment resistance.
We conducted an in vivo chemical screen to identify F1929-1458, which is capable of eliciting an organism-wide response to destroy stem cell tumors in Drosophila. We then performed functional validation using a mouse colorectal cancer graft tumor model established with the CT26 cell line characterized by its high content of CSCs. Single-cell sequencing was employed to analyze alterations in the TME. Small molecule pull-down mass spectrometry, cellular thermal shift assay, drug affinity experiment, and molecular docking were utilized to identify the target of F1929-1458. An in vitro co-culture system was applied to establish that the damage-associated molecular patterns (DAMPs) released by the tumor cells are accountable for the activation of dendritic cells (DCs).
We demonstrated that F1929-1458 treatment enhanced T cell infiltration and T cell mediated tumor regression, its anti-tumor effect was nullified in nude mice and was abolished after anti-CD3 neutralizing antibody treatment. We found that F1929-1458 binds NFKB1 to activate the NF-κB signaling pathway in tumor cells. The activation further elicits cellular stress, causing tumor cells to release DAMPs (HMGB1-gDNA complex, ATP, and OxLDL). These DAMPs, in turn, stimulate the cGAS-STING and NLRP3 inflammasome pathways in DCs, resulting in the generation of type I IFNs and IL-1β. These cytokines facilitate the maturation of DCs and antigen presentation, ultimately enhancing T cell-mediated anti-tumor immunity. Additionally, we showed that the combination of F1929-1458 and the anti-PD-1 antibody exhibited a synergistic anti-tumor effect.
Our study identified a novel NFKB1 agonist that promotes anti-tumor immunity by remodeling the TME and activating DCs and that may provide a new way to overcome resistance to current anti-tumor immunotherapy in colorectal cancer.
肿瘤微环境（TME）的免疫抑制特性以及癌症干细胞（CSC）的存在给肿瘤治疗带来了重大障碍。鉴定能够同时靶向CSC和TME的治疗药物可能是克服治疗抗性的一种潜在方法。
我们进行了一项体内化学筛选，以鉴定F1929 - 1458，它能够引发全生物体反应以破坏果蝇中的干细胞肿瘤。然后，我们使用以高CSC含量为特征的CT26细胞系建立的小鼠结直肠癌移植瘤模型进行功能验证。采用单细胞测序分析TME中的变化。利用小分子下拉质谱、细胞热迁移分析、药物亲和力实验和分子对接来鉴定F1929 - 1458的靶点。应用体外共培养系统来确定肿瘤细胞释放的损伤相关分子模式（DAMP）可导致树突状细胞（DC）的激活。
我们证明F1929 - 1458治疗增强了T细胞浸润和T细胞介导的肿瘤消退，其抗肿瘤作用在裸鼠中无效，并且在抗CD3中和抗体治疗后被消除。我们发现F1929 - 1458与NFKB1结合以激活肿瘤细胞中的NF - κB信号通路。这种激活进一步引发细胞应激，导致肿瘤细胞释放DAMP（HMGB1 - gDNA复合物、ATP和氧化低密度脂蛋白）。这些DAMP反过来刺激DC中的cGAS - STING和NLRP3炎性小体途径，导致I型干扰素和IL - 1β的产生。这些细胞因子促进DC的成熟和抗原呈递，最终增强T细胞介导的抗肿瘤免疫力。此外，我们表明F1929 - 1458与抗PD - 1抗体的联合使用表现出协同抗肿瘤作用。
我们的研究鉴定了一种新型的NFKB1激动剂，它通过重塑TME和激活DC来促进抗肿瘤免疫，并且可能为克服结直肠癌目前抗肿瘤免疫治疗的抗性提供一种新方法。
