DNMT3A inhibits E2F1-induced arterial marker expression and impairs angiogenesis in human umbilical artery endothelial cells

Abstract

Arterial marker genes EphrinB2 and HEY2 are essential for cardiovascular development and postnatal neovascularization. Our previous study confirmed that E2F1 could activate the transcription of EphrinB2 and HEY2 in human mesenchymal stem cells; however, the detailed mechanism has not been resolved yet. In this study, we focused on the interaction between E2F1 and DNMT3A, a de novo DNA methyltransferase, on regulating the expression of EphrinB2 and HEY2, and explored the potential mechanisms. Gain- and loss-of-function experiments implicated the positive effect of E2F1 on the expression of EphrinB2 and HEY2 and tube formation in human umbilical artery endothelial cells. Accumulation of DNMT3A decreased the levels of EphrinB2 and HEY2, and impaired tube formation induced by E2F1, while inhibiting DNMT3A by RNA interference augmented their expression and angiogenesis in E2F1-trasfected cells. We then asked whether the low expressions of EphrinB2 and HEY2 induced by DNMT3A are related to the methylation status of their promoters. Surprisingly, the methylation status of the CpG islands in the promoter region was not significantly affected by overexpression of exogenous DNMT3A. Furthermore, the interaction between E2F1 and DNMT3A was confirmed by co-immunoprecipitation. DNMT3A could inhibit the transcription of EphrinB2 and HEY2 promoters by affecting the binding of E2F1 to its recognition sequences as revealed by luciferase reporter assay and chromatin immunoprecipitation. These results identified a novel mechanism underlying the cooperation of DNMT3A with E2F1 on regulating target gene expression, and revealed their roles in the angiogenic process.

Pan-cancer analysis of DNMT3A as a robust prognostic biomarker to predict the immunotherapy response

“…Another related pathway is the transcription factor-related regulatory pathway. E2F is a famous family of transcription factors, which is involved in the regulation of multiple gene expression, and studies have shown that DNMT3A can participate in the biological phenotype regulated by E2F [55].…”

Potential epigenetic molecular regulatory networks in ocular neovascularization

“…In human umbilical artery DNMT3A has been shown to interact with E2F Transcription Factor 1 (E2F1) and control neovascularization-related genes such as the VEGF family, EPhinB2, and HEY2. However, the interaction of DNMT3A with E2F1 that modulates the gene's transcriptional repression may be independent of the methyltransferase activity (Su et al, 2020).…”