Large epigenome-wide association study of childhood ADHD identifies peripheral DNA methylation associated with disease and polygenic risk burden

<jats:title>Abstract</jats:title><jats:p>Epigenetic variation in peripheral tissues is being widely studied as a molecular biomarker of complex disease and disease-related exposures. To date, few studies have examined differences in DNA methylation associated with attention-deficit hyperactivity disorder (ADHD). In this study, we profiled genetic and methylomic variation across the genome in saliva samples from children (age 7–12 years) with clinically established ADHD (<jats:italic>N</jats:italic> = 391) and nonpsychiatric controls (<jats:italic>N</jats:italic> = 213). We tested for differentially methylated positions (DMPs) associated with both ADHD diagnosis and ADHD polygenic risk score, by using linear regression models including smoking, medication effects, and other potential confounders in our statistical models. Our results support previously reported associations between ADHD and DNA methylation levels at sites annotated to <jats:italic>VIPR2</jats:italic>, and identify several novel disease-associated DMPs (<jats:italic>p</jats:italic> < 1e–5), although none of them were genome-wide significant. The two top-ranked, ADHD-associated DMPs (cg17478313 annotated to <jats:italic>SLC7A8</jats:italic> and cg21609804 annotated to <jats:italic>MARK2</jats:italic>) are also significantly associated with nearby SNPs (<jats:italic>p</jats:italic> = 1.2e–46 and <jats:italic>p</jats:italic> = 2.07e–59), providing evidence that disease-associated DMPs are under genetic control. We also report a genome-wide significant association between ADHD polygenic risk and variable DNA methylation at a site annotated to the promoter of <jats:italic>GART</jats:italic> and <jats:italic>SON</jats:italic> (<jats:italic>p</jats:italic> = 6.71E–8). Finally, we show that ADHD-associated SNPs colocalize with SNPs associated with methylation levels in saliva. This is the first large-scale study of DNA methylation in children with ADHD. Our results represent novel epigenetic biomarkers for ADHD that may be useful for patient stratification, reinforce the importance of genetic effects on DNA methylation, and provide plausible molecular mechanisms for ADHD risk variants.</jats:p>