Methylation Files as Supplementary Material for Early Epigenetic Alterations in Cardiac Tissue Following Perinatal Sertraline Exposure in Mice

Targeted bisulfite sequencing (TBS-seq) profiled CpG methylation across predefined promoter and regulatory regions of 10 genes involved in serotonin signaling and cardiac function. Regional differential methylation was analyzed across 23 regions using overdispersion-corrected logistic regression with Benjamini–Hochberg (BH) adjustment. </p><p dir="ltr">Global 5-mC levels did not differ significantly between sertraline-exposed and control groups.

TBS-seq identified 1,941 CpG sites across 25 targeted regions. After coverage and regional quality filtering, 1,411 CpG sites across 23 regions were retained. Regional analysis identified two regions with significant differential methylation after FDR correction: the Htr2a enhancer showed an 8.19% increase in methylation (p<4.1x10-15, q<9.4x10-14), whereas the Tph2 promoter showed a 3.00% decrease in sertraline-exposed animals (p<0.0020, q<0.0229).

Secondary site-level analyses identified no individual CpG sites that remained significant after multiple-testing correction. Overall methylation profiles were highly correlated across samples, with no significant global difference between treatment groups. </p><p dir="ltr">Perinatal sertraline exposure was associated with localized regional DNA methylation changes in neonatal cardiac tissue without a global change in 5-mC.

The Htr2a enhancer and Tph2 promoter represent candidate regulatory loci linking early-life SSRI exposure with cardiac serotonergic pathways. These findings support further investigation of regional epigenetic alterations using integrated methylation, transcriptomic, and cell-type-resolved approaches to determine their functional significance and potential contribution to later-life cardiovascular outcomes.</p>