Cross-scale modeling reveals a TFRC-driven immunosuppressive macrophage niche in cervical cancer

Abstract

Background The functional plasticity of tumor-associated macrophages (TAMs) is a critical determinant of the immunosuppressive microenvironment in cervical cancer, yet its integration into actionable prognostic frameworks remains limited. This study aimed to establish a TAM polarization-centered model and elucidate the mechanisms of underlying tumor-immune crosstalk. Methods Bulk transcriptomics from The Cancer Genome Atlas (TCGA) were integrated with single-cell RNA sequencing (scRNA-seq) data (GSE208653). By combining weighted gene co-expression network analysis (WGCNA) with a multi-algorithm machine learning framework, a prognostic signature was constructed and independently validated in the Gene Expression Omnibus (GEO) GSE52903 cohort. Single-cell analysis resolved the cellular origins of signature genes, prioritizing tumor-enriched genes for validation. Protein-level expression was verified via immunohistochemistry (IHC) in a paired clinical cohort (n=39). Functional validation of the core gene was performed in vitro using cervical cancer cell lines co-cultured with THP-1-derived macrophages. Polarization was assessed via reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot (WB), enzyme-linked immunosorbent assay (ELISA), flow cytometry, and multiplex immunofluorescence (mIF). Results A robust five-gene prognostic signature (TP73, TFRC, SHC1, SCD, and PFKFB3) was developed, effectively stratifying patient survival. High risk scores correlated with a suppressed antitumor immune landscape and diminished predicted chemosensitivity to agents such as cisplatin. Single-cell analysis and IHC confirmed transferrin receptor (TFRC) as a tumor-intrinsic factor that is progressively upregulated during cervical carcinogenesis and enhances pro-M2 signaling. In vitro co-culture assays demonstrated that tumor-derived TFRC actively orchestrates an immunosuppressive M2-like macrophage niche, driving phenotypic shifts and pro-tumorigenic cytokine secretion, characterized by elevated interleukin-10 (IL-10) and reduced TNF-α. RT-qPCR analysis of 40 clinical specimens further confirmed a significant positive correlation between TFRC and the M2 marker Arg-1 at the mRNA level (r = 0.4961, P = 0.0011). Conclusions This study establishes a cross-scale, biologically interpretable prognostic model linking macrophage plasticity to clinical outcomes. We identify TFRC as a pivotal metabolic-immune node through which tumor-intrinsic iron metabolism orchestrates an immunosuppressive niche, providing a foundation for novel therapeutic strategies in cervical cancer.