Epigenetic profiling from stressed and non-stressed 3D cultured DNMT3A knock out cardiomyocytes.

This project was designed to evaluate the relevance of DNA methylation in human cardiomyocytes (CM).To this end, DNA methyl transferase DNMT3A was knocked out in human induced pluripotent stem cells (hiPSC) using CRISPR/Cas9 gene editing. Wild-type (WT) and knock out (KO) hiPSC were differentiated into CM. CM were used to create engineered heart tissue (EHT) in 24-well cell culture format. EHTs were cultured for 3 weeks. During the last week, culture was serum free and some EHTs were additionally stressed with 20 microM phenylephrine (PE) and 50 nM endothelin1 (ET1). DNA and RNA extracted from EHTs from all 4 groups (WT, WT PE/ET1, KO, KO PE/ET1) was then subjected to RNA sequencing and DNA methylation analysis by reduced representation bisulfite sequencing.