Multi-omics analysis of PFOA-induced sperm DNA damage in mice identifies candidate genes and mendelian randomization reveals their human genetic associations.

Perfluorooctanoic acid (PFOA) is a persistent environmental pollutant linked to male reproductive toxicity, particularly sperm DNA damage, yet the underlying molecular alterations remain incompletely understood. This study established a mouse model of PFOA exposure (10 mg/kg/day for 8 weeks), which significantly increased the sperm DNA fragmentation index. Integrated transcriptomic and proteomic analyses of testicular tissues identified 4781 differentially expressed genes and 639 differentially expressed proteins, with functional enrichment revealing pathways related to stress response, DNA repair, and metabolism. Four‑quadrant plot analysis pinpointed six molecules (ALDOC, FADS2, PNKD, RRP8, TAF9, and TRAPPC6A) exhibiting concordant dysregulation at both mRNA and protein levels. Mendelian randomization analysis further demonstrated significant genetic associations between these six genes and male infertility in humans. The dysregulation of five of these candidates was subsequently confirmed by qRT-PCR and/or Western blot, with ALDOC, TAF9, and TRAPPC6A showing consistent changes across all experimental approaches. These findings reveal that PFOA‑induced sperm DNA damage is associated with a complex molecular network and prioritize six candidate genes as targets for future functional investigation. This multi‑omics approach provides a comprehensive molecular landscape and offers refined targets for understanding PFOA reproductive toxicity.