Du Qiuyang, Ning Na, Zhao Xiujuan, Liu Feifan, Zhang Si, Xia Yuting, Li Fei, Yuan Shijie, Xie Xiaorong, Zhu Mengdi, Huang Zehan, Tang Zhaohui, Wang Jing, He Ran, Yang Xiang-Ping
Department of Immunology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
The Second Affiliated Hospital of Guangzhou Medical University, Guangdong Provincial Key Laboratory of Allergy & Clinical Immunology, Guangzhou 510260, China.
Theranostics. 2025 Jan 1;15(4):1304-1319. doi: 10.7150/thno.101298. eCollection 2025.
: Tumor associated macrophages (TAMs) are critical components in regulating the immune statuses of the tumor microenvironments. Although TAM has been intensively studied, it is unclear how mitochondrial proteins such as AGK regulate the TAMs' function. : We investigated the AGK function in TAMs using macrophage-specific deficient mice with B16 and LLC syngeneic tumor models. Flow cytometry was used to evaluate the stemness and activation of CD8 T cells. The enhanced release of mtDNA into the cytosol in the -deficient BMDMs was measured by RT-PCR and immunofluorescence; the cGAS-STING-type I IFN pathway was evaluated by immunoblotting. Mitochondria functions were evaluated by electron microscope and seahorse equipment. : We have noted an increased expression of AGK in TAMs of multiple tumor types, which was negatively correlates with the tumor tissue immune scores. In the B16 and LLC tumor models, macrophage -deficient mice have reduced tumor growth and enhanced populations of CD8 Tpex. AGK-deficient macrophages have increased mitochondrial damage and mtDNA release into the cytosol, which leads to enhanced cGAS-STING-type I IFN activation. Blockade of the type I IFN signaling pathway with anti-IFNAR reversed the phenotype in -deficient mice. : Our findings define a critical role of AGK in maintaining the macrophage mitochondrial homeostasis that is associated with mtDNA release and following cGAS-STING activation and type I IFN pathway. Targeting AGK in TAMs may represent a novel strategy to enhance anti-tumoral activity.
肿瘤相关巨噬细胞（TAM）是调节肿瘤微环境免疫状态的关键成分。尽管对TAM已进行了深入研究，但尚不清楚像AGK这样的线粒体蛋白如何调节TAM的功能。
我们使用B16和LLC同基因肿瘤模型的巨噬细胞特异性缺陷小鼠来研究AGK在TAM中的功能。采用流式细胞术评估CD8 T细胞的干性和活化情况。通过RT-PCR和免疫荧光检测AGK缺陷的骨髓来源巨噬细胞（BMDM）中mtDNA向细胞质的释放增强情况；通过免疫印迹评估cGAS-STING-I型干扰素途径。通过电子显微镜和海马设备评估线粒体功能。
我们注意到多种肿瘤类型的TAM中AGK表达增加，这与肿瘤组织免疫评分呈负相关。在B16和LLC肿瘤模型中，巨噬细胞AGK缺陷的小鼠肿瘤生长减缓，CD8 Tpex细胞群体增加。AGK缺陷的巨噬细胞线粒体损伤增加，mtDNA释放到细胞质中，导致cGAS-STING-I型干扰素活化增强。用抗IFNAR阻断I型干扰素信号通路可逆转AGK缺陷小鼠的表型。
我们的研究结果确定了AGK在维持与mtDNA释放以及随后的cGAS-STING活化和I型干扰素途径相关的巨噬细胞线粒体稳态中的关键作用。靶向TAM中的AGK可能代表一种增强抗肿瘤活性的新策略。
