Mechanisms of active DNA demethylation in human monocytes

The differentiation of human blood monocytes (MO), the post-mitotic precursors of macrophages (MAC) and dendritic cells (moDC), is accompanied by the active turnover of DNA methylation, but the extent, consequences and mechanisms of DNA methylation changes remain unclear. Here we profile and compare epigenetic landscapes during IL-4/GM-CSF-driven MO differentiation across the genome and detect several thousand regions that are actively demethylated during culture, both with or without accompanying changes in chromatin accessibility or transcription factor (TF) binding. We further identify TF that are globally associated with DNA demethylation processes. While interferon regulatory factor 4 (IRF4) is found to control hallmark DC functions with less impact on DNA methylation, early growth response 2 (EGR2), proves essential for MO differentiation as well as DNA methylation turnover at its binding sites. EGR2 is shown to interact with the 5mC hydroxylase TET2 and its consensus sequences show a characteristic DNA methylation footprint at demethylated sites with or without detectable protein binding. Our findings reveal a novel and essential role for EGR2 as epigenetic pioneer in human MO and suggest that active DNA demethylation can be initiated by TET2 recruiting TF both at stable and transient binding sites.EGA study EGAS00001004784