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

Sarah Haskell

doi:10.6084/m9.figshare.32509689

Maternal use of selective serotonin reuptake inhibitors (SSRIs), including sertraline, has been associated with an increased risk of congenital heart defects, but the underlying mechanisms remain unclear. SSRIs cross the placenta, directly exposing the developing fetus. Previous work demonstrated that perinatal sertraline exposure slows postnatal growth, suppresses calcium oscillations in cardiomyocytes, upregulates specific miRNAs, leading to downregulation of serotonin signaling components such as Htr2a, Htr2b, and Slc6a4, and chronically reduces left ventricular size. We hypothesized these effects may involve early epigenetic alterations in cardiac DNA methylation. Adult female C57BL/6 mice received daily intraperitoneal injections of sertraline (5 mg/kg) or saline for five days before mating and throughout gestation. Offspring continued to receive one-third of the maternal dose until postnatal day 14 (PN14), when hearts were collected for analysis. Global DNA methylation (5-methylcytosine, 5-mC) was quantified using a colorimetric ELISA assay. Targeted bisulfite sequencing (TBS-seq) was performed on extracted cardiac DNA to profile CpG methylation across promoter and regulatory regions of 10 genes implicated in serotonin signaling and cardiac function.Global 5-mC levels did not differ significantly between sertraline-exposed and control groups. However, TBS-seq analysis of 297 CpG sites identified 59 differentially methylated sites (p< 0.05), with 7 remaining significant after false discovery rate correction (p<0.01 and q< 0.05). These site-specific methylation changes were predominantly located within promoter regions of serotonin receptor genes (Htr2a, Htr2b, Htr4), the serotonin transporter gene (Slc6a4/SERT), and Tnnc1, a key regulator of cardiac contraction. Both hypermethylation and hypomethylation events were detected, often clustered in defined genomic regions, indicating complex, localized epigenetic remodeling. While global methylation remained stable, sertraline induced locus-specific epigenetic remodeling in neonatal cardiac tissue, notably in genes critical for serotonin signaling and calcium handling. These findings suggest early epigenetic programming as a potential mechanism underlying SSRI-associated alterations in cardiac development and function. Importantly, such site-specific epigenetic modifications could contribute to persistent changes in gene expression, ultimately influencing long-term cardiovascular health outcomes. This study highlights the importance of targeted epigenetic analyses to uncover subtle, gene-specific changes that may contribute to long-term cardiovascular outcomes following perinatal pharmacologic exposures.

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

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