Background Ferroptosis and immunogenic cell death play vital roles in colorectal cancer (CRC). The interplay between ferroptosis and immunogenic cell death (F-ICD) represents a promising frontier in cancer therapy. However, few studies have explored the combined regulatory effects of F-ICD in CRC. Methods In current study, we identified F-ICD related genes based on analysis of single-cell transcriptomics level and developed F-ICD related signature using 101 machine learning algorithms and WGCNA analysis. Differential analysis between normal and tumor samples was performed using DESeq2 (|logFC|>1, p. adj < 0.05). The RSF algorithm was chosen for further analysis due to its strong predictive performance, making it a robust tool for our study. An external validation was performed to access the expression level of seven key F-ICD related genes. Results By quantifying the expression levels of 44 genes related to F-ICD, we found that F-ICD activity was significantly elevated in NK cells, T cells, and some B cells. The module showed a significant correlation with the F-ICD score (r = 0.66). The predictive model had highly accurate AUCs in three datasets (0.99, 0.61, and 0.58 for the 3-years training sets), revealing the importance of F-ICD in different pathological stages and prognoses in CRC. Further results indicated that F-ICD was associated with pathways such as oxidative phosphorylation and NF-κB signaling. Patients with high F-ICD had significantly different mutation profiles and poorer prognoses. Conclusion This study developed a novel signature integrating ferroptosis and immunogenic cell death, creating a valuable model for predicting prognosis and the tumor immune environment in CRC. Furthermore, seven key genes emerged as promising targets for further investigation and therapeutic intervention, highlighting their potential role in ferroptosis and immunogenic cell death.

Objective: To delineate the expression profile and tumor-suppressive function of the metabolism-associated gene GPD1L in colorectal carcinogenesis. Methods: Transcriptomic datasets from TCGA and GEO repositories (GSE74602, GSE113513, GSE164191) were computationally analyzed. Paired tumor/adjacent mucosal specimens (n=58) from CRC patients at Jincheng People’s Hospital were analyzed alongside the NCM460 colon epithelial line and five CRC lines (SW620, HCT116, SW480, DLD-1, LOVO). Following GPD1L quantification via qPCR, selected cell models underwent pcDNA3.1-GPD1L transfection for functional characterization. Then Western blot analysis was used to explore its possible mechanism.Results: Comparative analysis revealed a marked elevation of GPD1L expression in non-neoplastic tissues relative to tumor specimens (P<0.001). Transcriptional profiling further identified significant depletion of GPD1L mRNA levels across malignant cell lines versus the NCM460 epithelial reference (P<0.05), with HCT116/SW620 showing maximal downregulation. Ectopic GPD1L expression attenuated oncogenic phenotypes: proliferation decreased (P<0.001), while Transwell quantification revealed 46.0% (HCT116: 605.0 ± 9.2 vs 326.7 ± 8.50 cells/field) and 54.3% (SW620: 455.3 ± 17.2 vs 208.0 ± 14.0 cells/field) reductions in migratory capacity (both P<0.001). Invasion assays showed parallel inhibition (HCT116: 43.3% decrease, P<0.01; SW620: 54.8% decrease, P<0.001). After overexpression of GPD1L, the expression levels of HIF-1α and MMP9 were reduced (P<0.05).Conclusion: GPD1L downregulation represents a hallmark of CRC progression, with affecting the expression of HIF-1α and MMP9 significantly impeding malignant behaviors, nominating it as a candidate tumor suppressor in colorectal neoplasia.