HK1–LDHA axis feedback regulation promotes histone lactylation of Wnt5a and mediates prenatal acetaminophen-induced susceptibility to osteoarthritis in male offspring

Acetaminophen is widely used during pregnancy but may cause developmental abnormalities in multiple systems in offspring. However, the effects of prenatal acetaminophen exposure (PAcE) on chondrodevelopment and long-term outcomes remain unclear. We administered 100 mg/kg per day of acetaminophen to rats on gestational days (GDs) 10-12 and treated fetal chondrocytes in vitro. In male PAcE offspring, cartilage matrix synthesis decreased and degradation increased at GD20 and postnatal week 12, with osteoarthritis (OA) susceptibility after running. Females exhibited only reduced matrix content at GD20. Mechanistically, acetaminophen inhibited the expression of hexokinase 1 (HK1) in chondrocytes under both normoxic and hypoxic conditions in vitro. However, HK1 inhibition induced feedback enhancement of the glycolytic only under hypoxic conditions, which increased lactic acid production and H3K18la lactylation levels in the Wnt5a promoter region, disrupting cartilage homeostasis. These effects were reversed by Wnt5a knockdown, HK1 overexpression, or LDHA knockdown. Finally, intra-articular adeno-associated virus-shWnt5a injection improved OA pathology in male PAcE offspring. In summary, PAcE upregulated H3K18la levels with Wnt5a through HK1-LDHA-mediated glycolytic feedback regulation, causing chondrodysplasia and increased OA susceptibility in male offspring, with sex differences. Our findings provide crucial insight into the hypoxia-dependent chondrotoxicity of PAcE, providing a reference for future research on fetal-derived OA. High H3K18la in the Wnt5a promoter region mediates PAcE-induced hypochondroplasia and susceptibility to adult OA in male offspring rats.