GenX-associated molecular signatures overlap with testicular aging and male infertility: a multi-omics integration analysis

GenX (HFPO-DA), a short-chain per- and polyfluoroalkyl substance (PFAS) substitute, is implicated in testicular toxicity. GenX-related genes were intersected with aging-associated genes to construct a GenX-Aging gene set. Single-cell RNA sequencing (scRNA-seq) data from human testicular aging (GSE254315) were analyzed to evaluate cell-type-specific aging sensitivity and intercellular communication dynamics. Male infertility transcriptomic datasets (GSE45885/GSE45887) were integrated, and least absolute shrinkage and selection operator (LASSO) regression combined with support vector machine recursive feature elimination (SVM-RFE) were applied to identify hub genes, which were validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR) in GenX-exposed rat testicular tissues. Spermatids exhibited the highest aging sensitivity, with progressive decline in intercellular communication. Four hub genes-SOD1, XRCC5, FOXO3, and POLB-demonstrated diagnostic value for male infertility. RT-qPCR confirmed computational predictions: SOD1, XRCC5, and FOXO3 were upregulated, while POLB was downregulated. Functional enrichment implicated FoxO signaling, cellular senescence, and DNA repair pathways. Molecular docking confirmed favorable GenX-protein binding interactions. SOD1, XRCC5, FOXO3, and POLB are candidate biomarkers for GenX-induced reproductive toxicity, with oxidative stress and genome maintenance as key pathological mechanisms.