Selective expansion of TCF7-expressing tumor-reactive T cell subpopulations during ovarian tumor-infiltrating T cell production ex vivo

期刊论文
发表日期：

2025年7月8日

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Dingfeng Liu,
Ting Zhang,
Qinli Sun,
Dongli Cai,
Yanan Lou,
Genyu Wang,
Bowen Xie,
Yicheng Zhu,
Chong Wang,
Zhouping Lu,
Chenfei Liu,
Yuan Li,
Xi Zhang,
Tianhui He,
Jing Hao,
Xinyi Guo,
Jiaming Li,
Xiaowei Xi,
Ling Ni,
Hongyan Guo,
Jing Ge,
Liping Jin,
Chen Dong

摘要

Tumor-infiltrating lymphocyte (TIL) therapy was recently approved for melanoma patients; however, the dynamic changes in T cell subpopulations during TIL production remain poorly understood. Here, we analyzed epithelial ovarian cancer samples at various stages of ex vivo TIL culture using paired single-cell RNA and TCR sequencing. We also assessed the expansion potential and tumor reactivity of the identified TIL subpopulations. Single-cell transcriptomic analysis revealed that CD8⁺ TILs exhibited reduced cellular diversity following ex vivo expansion, selectively expanding stem-like TCF7⁺ precursors of exhausted T cells (Tpex) and effector-like tissue-resident memory (Trm) cells. TCR clonotype analysis showed that Tpex cells accumulated through self-renewal, while Trm cells primarily originated from TCF7⁺GZMK⁺ early effector memory cells in tumors. Additionally, TCR tracing identified preferential activation and reprogramming of CD4⁺ T follicular helper (Tfh)-like cells, especially TCF7⁺ ones. All three TCF7⁺ subpopulations showed robust expansion potential and tumor reactivity in vitro. Notably, CCR7⁺CD200⁺ T cells, enriched for TCF-1⁺CD8⁺ Tpex and CD4⁺ Tfh-like cells in the tumor microenvironment, exhibited self-renewal during in vitro expansion and demonstrated tumor reactivity both in vivo and in vitro. These findings highlight the selective expansion of tumor-reactive TCF7⁺ T cells during TIL culture and suggest that CCR7 and CD200 serve as important surface markers for generating stem-like, tumor-reactive cells, potentially improving TIL therapy in cancers.