A mitoxyperilysis-related signature stratifies prognosis and identifies an aggressive colorectal cancer ecosystem with immune remodeling

Background Mitoxyperilysis is a recently described lytic cell death pathway linked to innate immune activation and metabolic disruption. Its clinical and biological relevance in colorectal cancer (CRC) remains unclear. This study aimed to develop a mitoxyperilysis-related signature (MRS) for prognostic stratification and to characterize its associated tumor microenvironmental features. Methods Bulk transcriptomic and clinical data from TCGA, GSE17536, and GSE29621 were used to construct and validate the MRS through an integrated machine-learning framework. Survival analysis, nomogram construction, pathway enrichment, immune deconvolution, ESTIMATE analysis, immunophenoscore-related assessment, and drug-response prediction were performed. Single-cell RNA sequencing and cell-cell communication analyses were used to define the cellular context of the MRS-associated phenotype. ANO1 was further evaluated by pan-cancer and CRC-specific analyses and validated by in vitro and in vivo experiments. Results The MRS consistently stratified overall survival across the training and validation cohorts and improved individualized risk prediction when incorporated into a nomogram. The MRS-high subtype was associated with extracellular matrix remodeling, invasion-related pathways, stromal enrichment, immune infiltration, and increased expression of inhibitory immune checkpoints. Single-cell analysis linked the adverse phenotype to epithelial cells with higher proliferative, metastatic, stress-adaptive, and immune-interactive features. These cells showed enhanced communication with stromal and immune compartments, particularly through FN1-, MK-, CXCL-, and CCL-related signaling. ANO1 was identified as a candidate downstream effector associated with the MRS-high state. Functionally, ANO1 knockdown suppressed CRC cell proliferation, migration, invasion, and xenograft tumor growth. Conclusion Mitoxyperilysis-related transcriptional programs are associated with a clinically aggressive and immune-remodeled ecosystem state in colorectal cancer. The MRS serves as a reliable tool for prognostic stratification and provides a transcriptional framework for characterizing CRC states linked to epithelial aggressiveness, microenvironmental remodeling, intercellular communication, and adverse clinical outcome. ANO1 may function as an important downstream effector associated with this MRS-high ecosystem state.