Single-cell transcriptomic analysis reveals gut microbiota-immunotherapy synergy through modulating tumor microenvironment

作者：Minyuan Cao, Yun Deng, Qing Hao, Huayun Yan, Q Wang, Chunyan Dong, Jing Wu, Yong He, Li‐Bin Huang, Xuyang Xia, Yongchao Gao, Hai‐Ning Chen, Weihan Zhang, Yanjing Zhang, Xiaozhen Zhuo, Lunzhi Dai, Hongbo Hu, Yong Peng, Feng Zhang, Zhaoqian Liu, Weihua Huang, Huiyuan Zhang, Yang Li, Yang Shu, Wei Zhang, Yan Zhang, Heng Xu · 发表于：Signal Transduction and Targeted Therapy · 年份：2025 · DOI：10.1038/s41392-025-02226-7 · 被引用次数：56 · 研究领域：Cancer Immunotherapy and Biomarkers、Gut microbiota and health、Immune cells in cancer

Abstract The gut microbiota crucially regulates the efficacy of immune checkpoint inhibitor (ICI) based immunotherapy, but the underlying mechanisms remain unclear at the single-cell resolution. Using single-cell RNA sequencing and subsequent validations, we investigate gut microbiota-ICI synergy by profiling the tumor microenvironment (TME) and elucidating critical cellular interactions in mouse models. Our findings reveal that intact gut microbiota combined with ICIs may synergistically increase the proportions of CD8 + , CD4 + , and γδ T cells, reduce glycolysis metabolism, and reverse exhausted CD8 + T cells into memory/effector CD8 + T cells, enhancing antitumor response. This synergistic effect also induces macrophage reprogramming from M2 protumor Spp1 + tumor-associated macrophages (TAMs) to Cd74 + TAMs, which act as antigen-presenting cells (APCs). These macrophage subtypes show a negative correlation within tumors, particularly during fecal microbiota transplantation. Depleting Spp1 + TAMs in Spp1 conditional knockout mice boosts ICI efficacy and T cell infiltration, regardless of gut microbiota status, suggesting a potential upstream role of the gut microbiota and highlighting the crucial negative impact of Spp1 + TAMs during macrophage reprogramming on immunotherapy outcomes. Mechanistically, we propose a γδ T cell-APC- CD8 + T cell axis, where gut microbiota and ICIs enhance Cd40lg expression on γδ T cells, activating Cd40 overexpressing APCs (e.g., Cd74 + TAMs) ...