Integrative Single-Cell and Bulk Transcriptomic Profiling Reveals Ferroptosis-Driven Heterogeneity of Ovarian Cancer Epithelial Cells

Abstract

Purpose: Ovarian cancer (OC) is a highly lethal gynecological malignancy characterized by insidious onset and poor prognosis, with limited validated prognostic biomarkers and therapeutic targets. Ferroptosis has been implicated in OC progression. However, most existing ferroptosis-related OC studies are based on bulk transcriptomic data, which fail to resolve the heterogeneity of OC epithelial cells. The relationship between ferroptosis-associated heterogeneity of epithelial cells and the tumor immune microenvironment remains insufficiently characterized at single-cell resolution. Materials and Methods: We integrated the GSE184880 single-cell RNA sequencing (scRNA-seq) discovery cohort, the independent GSE235329 scRNA-seq validation cohort, and bulk transcriptomic data from The Cancer Genome Atlas Ovarian Cancer (TCGA-OV) cohort to analyze epithelial cell subpopulations, ferroptosis activity, differentiation trajectories, and intercellular communication, and to construct a prognostic model. Results: OC epithelial cells were clustered into four subpopulations, which were further stratified into ferroptosis-high (EP_0 and EP_3) and ferroptosis-low (EP_1 and EP_2) groups. Key ferroptosis-associated epithelial patterns were reproduced in the independent GSE235329 scRNA-seq cohort. CellChat analysis identified MIF –( CD74 + CXCR4 ) and MIF –( CD74 + CD44 ) as prominent predicted interactions between OC epithelial cells and T cells. A validated three-gene prognostic model ( OGN , RARRES1 , IGFBP7 ) showed prognostic discrimination in the TCGA-OV cohort, with moderate predictive performance in the internal test set. Risk groups also differed in the xCell-estimated infiltration of selected immune and stromal cell populations. Conclusion: This study characterizes ferroptosis-associated heterogeneity of epithelial cells in OC at single-cell resolution and identifies candidate prognostic biomarkers and epithelial–immune communication signals that warrant further experimental and clinical validation.