scRNA-seq reveals that gestational HFPO-TA exposure impairs male reproductive function in offspring by inducing spermatocyte ferroptosis through the LCN2 axis.

Hexafluoropropylene oxide trimer acid (HFPO-TA) is widely used as an alternative to perfluorooctanoic acid (PFOA), yet the transgenerational toxicity and underlying molecular mechanisms of prenatal HFPO-TA exposure on the male reproductive system remain unclear. This study systematically investigated the toxic effects and mechanisms of HFPO-TA on the male reproductive system in offspring by establishing a mouse model of gestational HFPO-TA exposure (0.4 and 1 mg/kg/day) and integrating histopathology, single-cell transcriptomics, and molecular simulation technologies. The results demonstrated that HFPO-TA exposure significantly impaired the reproductive function of F1 male mice: testicular weight decreased, sex hormone levels declined, sperm quality compromised, spermatocyte meiotic arrest occurred, and apoptosis increased. Single-cell sequencing identified lipocalin-2 (Lcn2) as a key molecule mediating spermatocyte ferroptosis. Molecular docking and dynamics simulations confirmed stable binding between HFPO-TA and LCN2. In vitro experiments validated that HFPO-TA induced spermatocyte ferroptosis through lipid peroxidation and iron overload, which could be reversed by the ferroptosis inhibitor Fer-1 and Lcn2 overexpression. This study systematically revealed the reproductive toxicity mechanism whereby HFPO-TA induces spermatocyte-specific injury via the Lcn2-ferroptosis axis, providing a scientific basis for health risk assessment and regulation of next-generation PFAS.