Spatially Resolved Immunometabolic Reprogramming of Tumor-Associated Macrophages: An Integrative Multi-Layer Re-Analysis of Public Spatial Metabolomics, Spatial Transcriptomics and Single-Cell Resources

Background: Metabolic reprogramming of tumor-associated macrophages (TAMs) drives tumor immune evasion, yet the spatial and subtype-level organization of TAM metabolism remains under-characterized. Methods: We re-analyzed public spatial metabolomics, spatial transcriptomics, and single-cell datasets using autocorrelation-corrected spatial statistics, patient-level testing, and replication across breast, lung, colorectal, liver, and brain tumor data. Results: TAMs formed the most spatially clustered transcriptional module in breast tumor sections. TAM-dense regions were robustly coupled to iron handling (ρ = 0.568), surviving gene-overlap, cell-abundance, and erythrocyte/vascular-exposure controls, whereas apparent glycolysis and lipid-uptake links did not survive spatial or abundance correction. In lung adenocarcinoma, subtype comparisons were confounded by tissue origin; within tumor tissue, two differences persisted across patients (higher lipid uptake in C1QC⁺ and higher iron handling in FOLR2⁺ TAMs). A reported SPP1⁺-versus-C1QC⁺ polarity recurred at cell level across cancer types but not at patient level. Mitochondrial DNA tracing detected clonally expanded, metabolically hyperactive TAMs in two of four evaluable samples. Conclusions: TAM metabolism is spatially organized and subtype-specific, with iron handling as the most robust territorial coupling, supporting spatially stratified, subtype-aware design of metabolism-targeting trials.