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.