DNA methylation at CpG island shore and RXRα regulate NR2F2 in heart tissues of tetralogy of Fallot patients

Li Xiaodi; Ye Ming; Xu Hongfei; Zhu Yanjie; Gu Ruoyi; Xiaojing Ma; Sheng Wei; Huang Guoying

doi:10.1016/j.bbrc.2020.06.110

DNA methylation at CpG island shore and RXRα regulate NR2F2 in heart tissues of tetralogy of Fallot patients

Biochem Biophys Res Commun. 2020 Sep 3;529(4):1209-1215. doi: 10.1016/j.bbrc.2020.06.110. Epub 2020 Aug 7.

Authors

Li Xiaodi¹, Ye Ming², Xu Hongfei³, Zhu Yanjie¹, Gu Ruoyi², Xiaojing Ma², Sheng Wei⁴, Huang Guoying⁵

Affiliations

¹ Institute of Pediatrics, Children's Hospital of Fudan University, Shanghai, 201102, China; Shanghai Key Laboratory of Birth Defects, Shanghai, 201102, China.
² Cardiovascular Center, Children's Hospital of Fudan University, Shanghai, 201102, China.
³ Department of Forensic Medicine, Soochow University, Suzhou, 215006, China.
⁴ Institute of Pediatrics, Children's Hospital of Fudan University, Shanghai, 201102, China; Cardiovascular Center, Children's Hospital of Fudan University, Shanghai, 201102, China; Shanghai Key Laboratory of Birth Defects, Shanghai, 201102, China. Electronic address: sheng4616@sina.com.
⁵ Institute of Pediatrics, Children's Hospital of Fudan University, Shanghai, 201102, China; Cardiovascular Center, Children's Hospital of Fudan University, Shanghai, 201102, China; Shanghai Key Laboratory of Birth Defects, Shanghai, 201102, China. Electronic address: gyhuang@shmu.edu.cn.

PMID: 32819587
DOI: 10.1016/j.bbrc.2020.06.110

Abstract

The nuclear receptor subfamily 2 group F member 2 (NR2F2) gene encodes a ligand-inducible transcription factor involved in angiogenesis and heart development. This study aimed to elucidate the molecular mechanism of epigenetic regulation of NR2F2 in tetralogy of Fallot (TOF) development. In the present study, immunohistochemical staining showed that NR2F2 protein expression was significantly higher in the right ventricular outflow tract (RVOT) tissues of TOF cases compared with controls. The methylation status of the CpG island shore (CGIS) of the NR2F2 gene was decreased in TOF cases, and the CpG site 3 in the CGIS region of NR2F2 promoter was a differential methylation site. Furthermore, the methylation level of the CpG site 3 and the NR2F2 protein expression were significantly negatively correlated in TOF patients. In vitro functional analysis revealed that RXRα could upregulate the NR2F2 gene by directly binding to the CGIS in the NR2F2 promoter, while hypomethylation of the NR2F2 promoter via treatment with 5-azacytidine influenced the affinity of RXRα to its binding sites, as shown by ChIP-qPCR. These findings suggest that promoter hypomethylation activates NR2F2 by enhancing RXRα binding to NR2F2 CGIS in the development of TOF.

Keywords: CpG island shore; DNA methylation; NR2F2; RXRα; Tetralogy of Fallot.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adolescent
Animals
Base Sequence
COUP Transcription Factor II / genetics
COUP Transcription Factor II / metabolism*
Case-Control Studies
Child
Child, Preschool
CpG Islands / genetics*
DNA Methylation / genetics*
Female
HEK293 Cells
HeLa Cells
Humans
Male
Mice
Myocardium / metabolism*
Myocardium / pathology*
Promoter Regions, Genetic / genetics
Protein Binding
Reproducibility of Results
Retinoid X Receptor alpha / metabolism*
Tetralogy of Fallot / genetics*
Transcription, Genetic

Substances

COUP Transcription Factor II
NR2F2 protein, human
Retinoid X Receptor alpha