5mC profiling characterized TET2 as an anti-adipogenic demethylase

• Published in: Gene 2020-04, Vol.733, p.144265-144265, Article 144265
• Main Authors: Hou, Yingzi, Zhang, Zhenxi, Wang, Yao, Gao, Tao, Liu, Xiyang, Tang, Tao, Du, Quan
• Format: Article
• Language: English
• Subjects: 3T3-L1 Cells; 5-Methylcytosine - metabolism; Adipocytes - metabolism; Adipogenesis; Adipogenesis - genetics; Adipogenesis - physiology; Animals; Cell Differentiation - genetics; DNA Methylation; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Epigenesis, Genetic; Gene Expression - genetics; Lipolysis; Lipolysis - genetics; Mice; Promoter Regions, Genetic; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - metabolism; Receptors, Adrenergic, beta-3 - genetics; Receptors, Adrenergic, beta-3 - metabolism; TET2; Transcription Factors - genetics
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2019.144265

•TET2 is the most regulated 5mC demethylase during 3 T3-L1 differentiation.•TET2 is identified as an anti-adipogenic demethylase by loss-of function and gain-of function assays.•TET2 inhibits lipid accumulation by promoter demethylation of Adrb3.•Vitamin C was found to decrease 5mC level and repress lipid production. To explore its roles in adipogenesis, the levels of genomic 5mC methylation were examined across the adipocyte differentiation of 3 T3-L1 cells. This led to the identification of an up-regulating 5mC profile during the process. To further explore the regulation, gene expression assay was performed with a set of 5mC metabolic enzymes. Among them, TET2 was found to be the most regulated 5mC demethylase, in addition to a well-investigated 5mC methylase DNMT1. In the process, the expression of Tet2 increased for over 16-fold, suggesting its implications in the differentiation. Therefore, loss-of-function and gain-of-function assays were performed with Tet2. It was found that in relative to the differentiation of wild-type cells, knockdown of Tet2 expression led to greatly enhanced differentiation process, while over-expression of the gene resulted in repressed differentiation. Pathway study found that during the differentiation, TET2 demethylates Adrb3 promoter to up-regulate its expression. This led to enhanced lipolysis and decreased lipid production. To the upstream pathway, vitamin C treatment was found to enhance the activity of TETs, decrease 5mC levels and repress lipid production. Taken together, TET2 was characterized as an anti-adipogenic demethylase in adipocyte differentiation of 3 T3-L1 cells.