Metabolic reprogramming of cholesterol biosynthesis drives macrophage-mediated immune suppression in HPV-negative cervical adenocarcinoma

BackgroundCervical adenocarcinoma is an increasingly common and aggressive subtype of cervical cancer with marked biological heterogeneity. Accumulating evidence suggests that HPV-positive and HPV-negative adenocarcinomas exhibit distinct immune microenvironments, but the underlying mechanisms remain unclear.MethodsPublicly available single-cell RNA sequencing datasets of cervical adenocarcinoma and normal cervical tissues were systematically analyzed using integrated bioinformatic approaches, including cell clustering, copy number variation inference, metabolic pathway analysis, and cell–cell communication modeling.

Key findings were validated through in vitro experiments using cervical cancer cell lines, macrophage polarization assays, metabolic measurements, ELISA, immunofluorescence, and CD8+ T cell functional analyses.ResultsSingle-cell analysis revealed profound differences in cellular composition and immune states between HPV-negative and HPV-positive adenocarcinomas. HPV-negative tumors exhibited increased immune infiltration but were enriched for exhausted CD8+ T cells and immunosuppressive SPP1+ macrophages.

Malignant epithelial cells from HPV-negative adenocarcinoma displayed distinct metabolic reprogramming characterized by activation of cholesterol biosynthesis pathways, elevated DHCR7 expression, and accumulation of the oxysterol 27-hydroxycholesterol (27-HC). Functionally, 27-HC induced macrophage polarization toward an immunosuppressive phenotype and promoted SPP1 secretion.