Targeting thymidylate synthase enhances CD8 + T-cell infiltration and inhibits tumor growth in cervical cancer

Cervical cancer (CESC) presents a significant clinical challenge, primarily due to an incomplete understanding of the immunometabolic crosstalk within the tumor microenvironment (TME) and the lack of reliable biomarkers for immunotherapy stratification. Thymidylate synthase (TYMS), a pivotal enzyme in nucleotide synthesis, has been implicated in tumor progression, but its role as an immunometabolic regulator in CESC remains unexplored. Through an integrative approach combining single-cell RNA sequencing (scRNA-seq) of 47,589 cells, bulk transcriptomics, functional assays, and in vivo modeling, we delineated the multifaceted functions of TYMS. scRNA-seq analysis revealed a dynamic shift from CD4 + to exhausted CD8 + T-cell dominance during progression, orchestrated by specific ligand-receptor interactions like PTPRC-MRC1. A robust T cell-associated prognostic signature comprising TYMS, MYO6, SPINT1 and ESD was developed, effectively stratifying patients into distinct risk groups with differential tumor stemness, immune infiltration, and response to immunotherapy. Mechanistically, TYMS silencing promoted tumor stemness, migration, and invasion in vitro via targeting miR-197-3p. Crucially, in immunocompetent micemodel, TYMS knockdown accelerated tumor growth and potently suppressed CD8 + T-cell infiltration, demonstrating its role in promoting immune evasion. Conversely, TYMS overexpression suppressed tumorigenesis. Molecular docking identified Deoxyuridine Monophosphate as a high-affinity inhibitor of TYMS. Our findings demonstrate that targeting thymidylate synthase enhances CD8 + T-cell infiltration and inhibits tumor growth in cervical cancer, establishing TYMS as a promising therapeutic target.