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

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<sup>+</sup>, CD4<sup>+</sup>, and γδ T cells, reduce glycolysis metabolism, and reverse exhausted CD8<sup>+</sup> T cells into memory/effector CD8<sup>+</sup> T cells, enhancing antitumor response. This synergistic effect also induces macrophage reprogramming from M2 protumor Spp1<sup>+</sup> tumor-associated macrophages (TAMs) to Cd74<sup>+</sup> TAMs, which act as antigen-presenting cells (APCs). These macrophage subtypes show a negative correlation within tumors, particularly during fecal microbiota transplantation. Depleting Spp1<sup>+</sup> 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<sup>+</sup> TAMs during macrophage reprogramming on immunotherapy outcomes. Mechanistically, we propose a γδ T cell-APC-CD8<sup>+</sup> T cell axis, where gut microbiota and ICIs enhance Cd40lg expression on γδ T cells, activating Cd40 overexpressing APCs (e.g., Cd74<sup>+</sup> TAMs) through CD40-CD40L-related NF-κB signaling and boosting CD8<sup>+</sup> T cell responses via CD86-CD28 interactions. These findings highlight the potential importance of γδ T cells and SPP1-related macrophage reprogramming in activating CD8<sup>+</sup> T cells, as well as the synergistic effect of gut microbiota and ICIs in immunotherapy through modulating the TME.

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Publication Details

Journal
Signal Transduction and Targeted Therapy
Published
2025-05-01
DOI
https://doi.org/10.1038/s41392-025-02226-7
Citations
39
Primary Topic
Cancer Immunotherapy and Biomarkers
Type
article
Field-Weighted Citation Impact
40.77

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article

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

Huiyuan Zhang, Yanjing Zhang, Minyuan Cao, Wei Zhang et al.
39 citations
Signal Transduction and Targeted Therapy
Cancer Immunotherapy and Biomarkers
40.77
article

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

Huiyuan Zhang, Yanjing Zhang, Minyuan Cao, Wei Zhang, Yong He, Chunyan Dong, Yongchao Gao, Li‐Bin Huang, Xuyang Xia, Huayun Yan, Yang Shu, Weihua Huang, Hai‐Ning Chen, Zhaoqian Liu, Hongbo Hu, Qing Hao, Lunzhi Dai, Yong Peng, Feng Zhang, Yang Li, Yan Zhang, Yun Deng, Heng Xu, Jing Wu, Weihan Zhang, Q Wang, Xiaozhen Zhuo
article en
39 citations

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<sup>+</sup>, CD4<sup>+</sup>, and γδ T cells, reduce glycolysis metabolism, and reverse exhausted CD8<sup>+</sup> T cells into memory/effector CD8<sup>+</sup> T cells, enhancing antitumor response. This synergistic effect also induces macrophage reprogramming from M2 protumor Spp1<sup>+</sup> tumor-associated macrophages (TAMs) to Cd74<sup>+</sup> TAMs, which act as antigen-presenting cells (APCs). These macrophage subtypes show a negative correlation within tumors, particularly during fecal microbiota transplantation. Depleting Spp1<sup>+</sup> 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<sup>+</sup> TAMs during macrophage reprogramming on immunotherapy outcomes. Mechanistically, we propose a γδ T cell-APC-CD8<sup>+</sup> T cell axis, where gut microbiota and ICIs enhance Cd40lg expression on γδ T cells, activating Cd40 overexpressing APCs (e.g., Cd74<sup>+</sup> TAMs) through CD40-CD40L-related NF-κB signaling and boosting CD8<sup>+</sup> T cell responses via CD86-CD28 interactions. These findings highlight the potential importance of γδ T cells and SPP1-related macrophage reprogramming in activating CD8<sup>+</sup> T cells, as well as the synergistic effect of gut microbiota and ICIs in immunotherapy through modulating the TME.

Signal Transduction and Targeted TherapyVol. 10(1)
Central South University (CN), Sichuan University (CN), Wenzhou Medical University (CN), Quzhou City People's Hospital (CN), West China Hospital of Sichuan University (CN), Hunan Cancer Hospital (CN), Sichuan Cancer Hospital (CN), First Affiliated Hospital of Guangdong Pharmaceutical University (CN), First Affiliated Hospital of Xi'an Jiaotong University (CN), Xiangya Hospital Central South University (CN), State Key Laboratory of Biotherapy
National Natural Science Foundation of China, China Postdoctoral Science Foundation, Sichuan University, West China Hospital, Sichuan University
Good health and well-being
Openalex Percentile: Top 0%
Cancer Immunotherapy and Biomarkers
40.77
DOIOpen Access PDFOpenAlex
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