Unraveling CD4+ T cell heterogeneity and cell death-associated genes in high-grade serous ovarian cancer: a comprehensive analysis of single-cell RNA and spatial transcriptome sequencing

• Shaobo Wu ^[1] ; Chunfeng Wu ^[1] ; Jiayi Yu ^[1] ; Yaoyao Li ^[1] ; Yanhong Lv ^[1] ; Lei Ding ^[1] ; Di Zhang ^[1] ; Yurou Zhou ^[2] ; Jun Tang ^[1]
  1. [1] Department of Laboratory Medicine, Tinglin Hospital of Jinshan District, Shanghai, 201505, People’s Republic of China
  2. [2] Jinshanwei Town Community Healthcare Center, Jinshan District, Shanghai, 201512, People’s Republic of China
• Localización: Clinical & translational oncology, ISSN 1699-048X, Vol. 28, Nº. 8, 2026, págs. 3253-3276
• Idioma: inglés
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• Resumen

  • Background:

    CD4+ T cells infiltrate the tumor microenvironment, but their role in high-grade serous ovarian cancer (HGSOC) progression remains unclear. Cell death-associated genes critically impact HGSOC advancement.

    Methods:

    We integrated single-cell and spatial transcriptomics to analyze CD4+ T cell heterogeneity and identify cell deathassociated genes. Key candidates were experimentally validated via qPCR and Western blotting in independent HGSOC cohorts. Furthermore, a T cell–tumor cell co-culture system was employed for functional validation of candidate gene effects on tumor behavior.

    Results:

    To comprehensively characterize the heterogeneity of CD4+ T cells in HGSOC, we performed an integrated analysis including inferCNV, pseudotime trajectory, SCENIC transcription regulatory networks, and hdWGCNA. This multi-omics approach defined distinct CD4+ T cell subgroups and identified key regulons. Notably, a co-expression network module derived from hdWGCNA was leveraged to construct a prognostic model, which demonstrated significant predictive power for patient survival. Spatial transcriptomic analysis identified co-localized niches of IL7R+CD4+ T cells and Tregs, exhibiting highly concordant gene expression with scRNA-seq data. Orthogonal validation confirmed significant dysregulation of MAL (p<0.001) and ANXA1 (p<0.001) in HGSOC. Critically, in vitro co-culture experiments demonstrated that silencing MAL in T cells significantly suppressed ovarian cancer cell proliferation, migration, and invasion, highlighting its functional role in modulating the tumor microenvironment.

    Conclusion:

    Our findings elucidate mechanisms of cell death-associated genes in CD4+ T cell subgroups, with experimentally grounded biomarkers offering therapeutic options for HGSOC.