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 ova...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.展开更多
Although PD-L1/PD-1 blockade therapy has been approved to treat many types of cancers,the majority of patients with solid tumors do not respond well,but the underlying reason remains unclear.Here,we studied ovarian ca...Although PD-L1/PD-1 blockade therapy has been approved to treat many types of cancers,the majority of patients with solid tumors do not respond well,but the underlying reason remains unclear.Here,we studied ovarian cancer(OvCa),a tumor type generally resistant to current immunotherapies,to investigate PD-1-independent immunosuppression.We found that PD-L1 was not highly expressed in the tumor microenvironment(TME)of human OvCa.Instead,B7-H3,another checkpoint molecule,was highly expressed by both tumor cells and tumor-infiltrating antigen-presenting cells(APCs),which correlated with T-cell exhaustion in patients.Using ID8 OvCa mouse models,we found that B7-H3 expressed on tumor cells,but not host cells,had a dominant role in suppressing antitumor immunity.Therapeutically,B7-H3 blockade,but not PD-1 blockade,prolonged the survival of ID8 tumorbearing mice.Collectively,our results demonstrate that tumor-expressed B7-H3 inhibits the function of CD8^(+)T cells and suggest that B7-H3 may be a target in patients who are not responsive to PD-L1/PD-1 inhibition,particularly OvCa patients.展开更多
基金supported by the National Key Research and Development Program of China(2021YFC2302403,2022YFC2704703,2022YFC2702204,2024YFA1306001)the National Natural Science Foundation of China(31991173,31991170,82271701,81730039,82071653,82102853,82372888)+2 种基金the Shanghai Pudong New Area Health Commission Project(PW2022D-05)Peking University Clinical Scientist Training Program of the Fundamental Research Funds for the Central Universities(BMU2024PYJH008)supported by a New Cornerstone Investigator award.
文摘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.
基金The authors are grateful to Dr.Xueguang Zhang(Soochow University,China)and Jiajia Li(Fudan University Shanghai Cancer Center)for providing the mouse and human OvCa cell lines.This project was funded in part by the Key Program of the National Natural Science Foundation of China(81730039&81671460 to L.-P.J.)the National Key Research and Development Program of China(2017YFC1001401 to L.-P.J.)+2 种基金Beijing Municipal Science and Technology Projects(Z181100006318015 and Z181100001318007 to C.D.)Shanghai Municipal Medical and Health Discipline Construction Projects(2017ZZ02015 to L.-P.J.)the National Basic Research Program of China(2015CB943300 to L.-P.J.).
文摘Although PD-L1/PD-1 blockade therapy has been approved to treat many types of cancers,the majority of patients with solid tumors do not respond well,but the underlying reason remains unclear.Here,we studied ovarian cancer(OvCa),a tumor type generally resistant to current immunotherapies,to investigate PD-1-independent immunosuppression.We found that PD-L1 was not highly expressed in the tumor microenvironment(TME)of human OvCa.Instead,B7-H3,another checkpoint molecule,was highly expressed by both tumor cells and tumor-infiltrating antigen-presenting cells(APCs),which correlated with T-cell exhaustion in patients.Using ID8 OvCa mouse models,we found that B7-H3 expressed on tumor cells,but not host cells,had a dominant role in suppressing antitumor immunity.Therapeutically,B7-H3 blockade,but not PD-1 blockade,prolonged the survival of ID8 tumorbearing mice.Collectively,our results demonstrate that tumor-expressed B7-H3 inhibits the function of CD8^(+)T cells and suggest that B7-H3 may be a target in patients who are not responsive to PD-L1/PD-1 inhibition,particularly OvCa patients.
