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Early life stress has been linked to increased methylation of the Nuclear Receptor Subfamily 3 Group C Member 1 ( NR3C1 ) gene, which codes for the glucocorticoid receptor. Moreover, early life stress has been associated with substance use initiation at a younger age, a risk factor for developing substance use disorders. However, no studies to date have investigated whether  NR3C1  methylation can predict substance use in young individuals. This study included adolescents 13–14 years of age that reported no history of substance use at baseline, ( N  = 1041; males = 46%). Participants contributed saliva DNA samples and were followed in middle adolescence as part of KUPOL, a prospective cohort study of 7th-grade students in Sweden. Outcome variables were self-reports of (i) recent use, (ii) lifetime use, and (iii) use duration of (a) alcohol, (b) tobacco products, (c) cannabis, or (d) any substance. Outcomes were measured annually for three consecutive years. The predictor variable was DNA methylation at the exon 1 F locus of  NR3C1 . Risk and rate ratios were calculated as measures of association, with or without adjustment for internalizing symptoms and parental psychiatric disorders. For a subset of individuals ( N  = 320), there were also morning and afternoon salivary cortisol measurements available that were analyzed in relation to  NR3C1  methylation levels. Baseline  NR3C1  hypermethylation associated with future self-reports of recent use and use duration of any substance, before and after adjustment for potential confounders. The overall estimates were attenuated when considering lifetime use. Sex-stratified analyses revealed the strongest association for cigarette use in males. Cortisol analyses revealed associations between  NR3C1  methylation and morning cortisol levels. Findings from this study suggest that saliva  NR3C1  hypermethylation can predict substance use in middle adolescence. Additional longitudinal studies are warranted to confirm these findings.

DNA methylation of the glucocorticoid receptor gene predicts substance use in adolescence: longitudinal data from over 1000 young individuals