小 G 蛋白 Arf1 已被确定在支持癌症干细胞 (CSC) 方面发挥选择性作用，使其成为癌症治疗的一个有吸引力的靶标。然而，目前的Arf1抑制剂由于其高毒性和低特异性，其转化潜力有限。在这项研究中，引入了两种新型强效 Arf1 小分子抑制剂，分别为 DU101 和 DU102，用于癌症治疗。临床前肿瘤模型表明，这些抑制剂引发了 CSC 的级联衰老，并增强了小鼠癌症和 PDX 模型的抗肿瘤免疫力。通过单细胞测序，分析了这些新型 Arf1 抑制剂诱导的肿瘤免疫微环境的重塑，并确定了肿瘤相关 CD8 CD4 双阳性 T (DPT) 细胞的增加。这些 DPT 细胞表现出活性 CD8 单阳性 T 细胞的卓越特征，以及较高比例的 TCF1 PD-1（干样 T 细胞的特征）。肿瘤浸润干细胞样 DPT 细胞的频率与癌症患者更好的无病生存 (DFS) 相关，表明这些抑制剂可能通过将冷的肿瘤免疫微环境转变为热的免疫微环境来提供一种新的癌症免疫治疗策略，从而扩展癌症患者免疫治疗的潜力。© 2024 作者。 《Advanced Science》由 Wiley‐VCH GmbH 出版。

The small G protein Arf1 has been identified as playing a selective role in supporting cancer stem cells (CSCs), making it an attractive target for cancer therapy. However, the current Arf1 inhibitors have limited translational potential due to their high toxicity and low specificity. In this study, two new potent small-molecule inhibitors of Arf1, identified as DU101 and DU102, for cancer therapy are introduced. Preclinical tumor models demonstrate that these inhibitors triggered a cascade of aging in CSCs and enhance anti-tumor immunity in mouse cancer and PDX models. Through single-cell sequencing, the remodeling of the tumor immune microenvironment induced by these new Arf1 inhibitors is analyzed and an increase in tumor-associated CD8+ CD4+ double-positive T (DPT) cells is identified. These DPT cells exhibit superior features of active CD8 single-positive T cells and a higher percentage of TCF1+PD-1+, characteristic of stem-like T cells. The frequency of tumor-infiltrating stem-like DPT cells correlates with better disease-free survival (DFS) in cancer patients, indicating that these inhibitors may offer a novel cancer immunotherapy strategy by converting the cold tumor immune microenvironment into a hot one, thus expanding the potential for immunotherapy in cancer patients.© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.