Exploratory ferroptosis-related transcriptomic patterns in dilated cardiomyopathy and pan-cancer datasets with H9c2 RT-qPCR assessment

Abstract

Ferroptosis-related pathways have been implicated in cardiovascular remodeling and cancer, but their relevance to hypertensive heart disease (HHD) remains uncertain. This study examined transcriptomic patterns in a dilated cardiomyopathy (DCM) dataset and used pan-cancer data only for descriptive context; it did not analyze primary HHD tissue. GSE183852 data were restricted to the explicitly identified non-diseased sample H_ZC-11-292 (8,749 cells) and DCM sample HDCM1 (7,202 cells). H_ZC-LVAD-1 was excluded because GEO metadata classify it as DCM. Full-rank cell-level Wilcoxon results were retained, with all single-cell tests treated as descriptive. GSE5406 groups were assigned from GEO sample titles (194 failing and 16 non-failing myocardium samples), and TCGA data were used for descriptive pan-cancer context. GPX4, SLC7A11, ACSL4, and TFRC mRNA were assessed by RT-qPCR in Ang II-stimulated H9c2 cells. Fourteen cluster-derived cell categories were retained. Cardiomyocytes, macrophages, and endothelial cells were not detected in the selected DCM donor, so no condition-specific inference was made for those categories. Among 109 detected ferroptosis-related genes, 60 met the cell-level up threshold, 24 met the down threshold, and 25 were nonsignificant; these counts are not donor-level evidence. In GSE5406, 5 of 99 tested ferroptosis-related genes met |log2FC| > 0.5 and BH-adjusted p < 0.05. Fold changes for 96 detected genes were uncorrelated across platforms (r = 0.034, p = 0.740). RT-qPCR showed lower GPX4 and SLC7A11 and higher ACSL4 and TFRC mRNA, with gene- and context-specific directionality (Supplementary Table S1). The analyses identify exploratory, donor- and context-dependent transcriptional patterns. They do not establish ferroptosis as a causal mechanism, biomarker, or treatment target in primary HHD. Multi-donor disease-specific cohorts and protein-level and functional experiments are required.