Integrated multi-omics identifies a CD54+ iCAF-ITGAL+ macrophage niche driving immunosuppression via CXCL8-PDL1 axis in cervical cancer.

Abstract

Cervical cancer (CC) remains a formidable clinical challenge, particularly in advanced stages where immune checkpoint blockade yields suboptimal responses. Despite the established role of the tumor microenvironment (TME) in fostering immunosuppression, the precise mechanisms of stroma-immune crosstalk in CC remain elusive. Leveraging single-cell RNA sequencing of 77,221 cells from CC and normal cervical tissues, we uncovered a tumor-enriched subpopulation of inflammatory cancer-associated fibroblasts (iCAFs) marked by elevated CD54 expression (CD54+ iCAFs), which independently predicted adverse clinical outcomes. Systematic dissection of intercellular communication networks revealed a tumor-specific alliance between CD54+ iCAFs and ITGAL+ macrophages, orchestrated through dysregulated ligand-receptor signaling. Spatial multi-omics approaches, including multiplex immunohistochemistry and spatial transcriptomics, confirmed their colocalization within an immunosuppressive niche. Mechanistically, CD54+ iCAFs promote immunosuppression by polarizing ITGAL+ macrophages toward an M2-like phenotype, primarily via CCL2 secretion. These fibroblasts further support immune evasion through two complementary pathways: direct CD54-ITGAL contact-dependent signaling and soluble CCL2-mediated macrophage reprogramming. The resulting macrophage activation stimulates autocrine CXCL8 secretion and subsequent PD-L1 upregulation, which ultimately suppresses CD8+ T cell functions, fostering an immune-tolerant microenvironment in CC. Therapeutic intervention using the CXCL8-CXCR1/2 inhibitor reparixin disrupted the CXCL8-PD-L1 axis, reduced PD-L1+ macrophage abundance and enhanced CD8+ T cell cytotoxicity. Notably, combination therapy with PD-L1 blockade demonstrated synergistic efficacy. Collectively, our findings reveal a stromal-immune checkpoint axis orchestrated by CD54⁺ iCAFs and ITGAL⁺ macrophages that underpins immunosuppression in CC, thereby providing a translational rationale for stroma-directed combination therapies that may overcome resistance to current immunotherapies.