| Summary: | Epigenetic changes are associated with altered behavior and neuropsychiatric disorders and they modify the trajectory of aging. Maternal anxiety during pregnancy is a common environmental challenge for the fetus, causing changes in DNA methylation. Here, we determined the mediating role of DNA methylation and the moderating role of offspring sex on the association between maternal anxiety and children’s behavioral measures. In 83 mother–child dyads, maternal anxiety was assessed in each trimester of pregnancy when the child was four years of age. Children’s behavioral measures and children’s buccal DNA methylation levels (<i>NR3C1</i>, <i>IGF2/H19 ICR</i>, and <i>LINE1</i>) were examined. Higher maternal anxiety during the third trimester was associated with more methylation levels of the <i>NR3C1</i>. Moderating effects of sex on the association between maternal anxiety and methylation were found for <i>IGF2/H19</i> and <i>LINE1</i> CpGs. Mediation analysis showed that methylation of <i>NR3C1</i> could buffer the effects of maternal anxiety on children’s behavioral measures, but this effect did not remain significant after controlling for covariates. In conclusion, our data support an association between maternal anxiety during pregnancy and DNA methylation. The results also underscore the importance of sex differences and timing effects. However, DNA methylation as underlying mechanism of the effect of maternal anxiety during pregnancy on offspring’s behavioral measures was not supported.
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