Glutathione metabolism-linked ferroptosis in human seminoma: a spatial multi-omics mapping study

Testicular cancer is the most common solid tumor in young men aged 15-44 years, with seminoma being a major subtype. To elucidate its poorly understood molecular mechanisms, we performed integrated spatial transcriptomics (ST) and spatial metabolomics (SM) on fresh-frozen seminoma tissues from four patients. ST identified 16 distinct cell clusters and spatially resolved transcriptional programs in tumor, invasive, tubular, and stromal regions, revealing tumor-enriched pathways like glycolysis, oxidative phosphorylation, and ferroptosis. Pseudotime analysis suggested a branched trajectory of malignant progression. SM showed pronounced dysregulation of glutathione metabolism in tumor areas. Integrated analysis pinpointed glutathione metabolism and ferroptosis as convergent regulatory pathways in seminoma. Functional validation in TCam-2 cells and a xenograft mice demonstrated that pharmacologic inhibition of glutathione synthesis (using buthionine sulfoximine, BSO) or genetic knockdown of xCT induced ferroptosis and suppressed tumor cell growth both in vitro and in vivo. This study establishes glutathione metabolic reprogramming as a key adaptive mechanism in seminoma that maintains redox homeostasis and protects against ferroptosis, suggesting that targeting this pathway or promoting ferroptosis represents a novel therapeutic strategy.