Abstract
Tumor-Infiltrating Lymphocytes (TILs) immunotherapy is a highly promising treatment for Non-small Cell Lung Cancer (NSCLC), which is responsible for 18% of all cancer-related deaths. The heterogeneity of TILs remains poorly understood. Here, we utilized combined single-cell RNA (scRNA)/T cell receptor sequencing (scTCR-seq) data from lung adenocarcinoma (LUAD) patients. Naïve CD4+ and effector memory CD8+ T cells were increased in tumor tissue compared with circulating blood samples. Activated signaling pathways were detected, and GZMA was identified as a potential novel diagnostic biomarker. During the transitional phase, macrophages (FTL) and dendritic (AIF1) cells transported the most CD3 TCR clones to T cells, while cytotoxicity CD8+ T (NKG7) cells transported to terminal exhausted CD8+ T cells. In both transition and expansion phases, T helper cells (CXCL13) are transported to regulatory T cells (Tregs). Additionally, we investigated the expression profiles of key cytokines, checkpoint receptors, and their ligands. Cytotoxicity CD8+ T cells (CCL5 and IFNG), T helper cells (FTL, TNFRSF4, and TIGIT), and regulatory T cells (CTLA4, TIGIT and FTL) exhibited functional roles in both primary and metastatic tumor stages. Taken together, our study provides a single-cell resolution of the TIL immune landscape and suggests potential treatment strategies to overcome drug resistance.
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Data availability
A comprehensive survey of the literature on next-generation sequencing (NGS) studies involving TILs in NSCLC identified three recent publications: 1. Gueguen et al., referred to as “scRNA-seq and scTCR-seq data of resident and circulating precursors to tumor-infiltrating CD8+ T cell populations in lung cancer” (GSE162498 and GSE162499) [18]. 2. Laughney et al., based on the “scRNA-seq transcriptional landscape of primary tumors and metastases human LUAD” (GSE123902) [39]. 3. Ganesan et al., referred to as “Analysis of purified populations of CD8 T cells (isolated from primary lung tumors and matched adjacent lung tissue of lung cancer patients) at the transcriptomic level by RNA sequencing” (GSE90728) [40].
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Acknowledgements
We thank everyone who contributed to this study and the GEO database for the analytical data.
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JGL analyzed scRNA-seq data. LY analyzed scTCR-seq data. XLG performed statistical analysis. XMH performed ligand–receptor analysis. ML conducted trajectory analysis. RYG validated data. BHZ performed Kaplan–Meier analysis. QYL generated GraphPad plots. WJZ prepared figures. PX annotated cell types. XHG analyzed correlations. YAC prepared T cell figures. XLY drafted the manuscript. YS approved the final version. ZHG provided immunological expertise. JHM edited the language. YXH improved image resolution. LML assisted cancer immunology. JH supported funding. HZ conceived the project. YL supervised the study. All authors reviewed and approved the final manuscript.
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This study did not require ethical approval as it did not involve human participants, human data, or animal subjects. All analyses were based on publicly available, anonymized datasets (e.g., UALCAN, GEO, GEPIA, Kaplan–Meier) that had obtained appropriate ethical clearance prior to release and are openly accessible to the research community. Therefore, no institutional ethics review was necessary.
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Liu, JG., Yu, L., Guo, XL. et al. Characterizing the immune landscape of tumor-infiltrating lymphocytes in non-small cell lung cancer. Genes Immun 26, 229–241 (2025). https://doi.org/10.1038/s41435-025-00330-w
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DOI: https://doi.org/10.1038/s41435-025-00330-w
