Integrative analysis of single-cell and bulk RNA sequencing data for discovery of senescent TAMs prognostic characteristics in neuroblastoma.

Tumor-associated macrophages (TAMs) comprise heterogeneous subtypes with context-dependent functions in tumor immunity. Although macrophage senescence influences diverse diseases, its role in neuroblastoma (NB) progression remains undefined. Single-cell RNA sequencing (scRNA-seq) data from NB samples (GEO: GSE147766) were analyzed to identify TAM subtypes. Senescence-associated signatures were characterized via pseudotime trajectory, cell-cell communication, and functional assays. A prognostic model was developed using Cox/LASSO regression and validated in independent cohorts (TARGET-NBL, E-MTAB-8248). Drug sensitivity and immune microenvironment differences were compared between risk groups. In vitro validation assessed EIF5's role in TAM senescence and NB progression. We identified a senescent TAM subset (C0) exhibits a hybrid M1/M2 state with a dominance of M2-like features. Pseudotime analysis revealed differentiation from non-senescent TAMs (C1), accompanied by upregulated LILRB1, LILRB2, IL10, and CXCL8. A 7-gene prognostic signature (AMD1, ARL4A, BRD2, DDX3X, EIF5, SLC43A2, ZEB2) stratified patients into risk groups with distinct survival (P < 0.001). High-risk patients displayed impaired anti-tumor immunity (reduced CD8⁺ T cells, elevated TIDE scores) and differential drug sensitivity. EIF5 knockdown suppressed senescence markers (p21/p16) in macrophages and inhibited NB cell proliferation/migration. Senescent TAMs influence NB immunosuppression and progression. EIF5 is a key regulator of TAM senescence and a potential therapeutic target. Our risk model may guide clinical stratification and targeted interventions.