Examination of DNA Methylation Patterns in Children Born Premature with Prenatal Tobacco Smoke Exposure.

Prenatal tobacco smoke exposure (TSE) has been associated with significant alterations in DNA methylation (DNAm), an epigenetic mechanism with potential functional consequences to child development. This pilot study aimed to investigate differential DNAm patterns in preterm children with and without prenatal TSE using reduced representation bisulfite sequencing (RRBS) to interrogate a wider array of sites than in more common approaches, namely microarrays. Buccal swabs were collected from 16 two-year-old children (7 with TSE, 9 without), and DNAm was quantified at over 1.3 million CpG sites. To identify differential DNAm, univariable analyses were first performed and followed by Bayesian beta-binomial hierarchical regression models for sequence count data including adjustment for potential confounders. False Discovery Rate correction was used to account for multiple comparisons. Significant differential methylation was observed at CpG sites within intronic regions of the CALN1 and LINGO1 genes and the distal intergenic region of the TBL1XR1 gene. These findings suggest that prenatal TSE may influence epigenetic regulation in genes involved in neurodevelopment. This study demonstrates the importance of RRBS in identifying novel DNAm changes associated with prenatal TSE and highlights the need for larger studies to validate and expand upon these preliminary findings.