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Gene. 2017 Nov 30;635:48-60. doi: 10.1016/j.gene.2017.09.006. Epub 2017 Sep 6.

Integrative analysis of DNA methylation and mRNA expression during differentiation of umbilical cord blood derived mononuclear cells to endothelial cells.

Author information

1
Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea.
2
Department of Life Science, Ewha Research Center for Systems Biology (ERCSB), Ewha Womans University, Seoul, Republic of Korea. Electronic address: yukyungjun@ewha.ac.kr.
3
Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
4
Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon 200-701, Republic of Korea.
5
Department of Life Science, Ewha Research Center for Systems Biology (ERCSB), Ewha Womans University, Seoul, Republic of Korea.
6
Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749, Republic of Korea. Electronic address: ygkwon@yonsei.ac.kr.

Abstract

Differentiation of umbilical cord blood derived mononuclear cells to endothelial cells is accompanied by massive changes in gene expression. Although methylation and demethylation of DNA likely play crucial roles in regulating gene expression, their interplay during differentiation remains elusive. To address this question, we performed deep sequencing of DNA methylation and mRNA expression to profile global changes in promoter methylation and gene expression during differentiation from mononuclear cells to outgrowing cells. We identified 61 downregulated genes with hypermethylation, including CD74, VAV1, TLR8, and NCF4, as well as 21 upregulated genes with hypomethylation, including ECSCR, MCAM, PGF, and ARHGEF15. Interestingly, gene ontology analysis showed that downregulated genes with hypermethylation were enriched in immune-related functions, and upregulated genes with hypomethylation were enriched in the developmental process and angiogenesis, indicating the important roles of DNA methylation in regulating differentiation. We performed polymerase chain reaction analyses and bisulfite sequencing of representative genes (CD74, VAV1, ECSCR, and MCAM) to verify the negative correlation between DNA methylation and gene expression. Further, inhibition of DNA methyltransferase and demethylase activities using 5'-aza-dc and shRNAs, specific for TET1 and TET2 mRNAs, respectively, revealed that DNA methylation was the main regulator of the reversible expression of functionally important genes. Collectively, our findings implicate DNA methylation as a critical regulator of gene expression during umbilical cord blood derived mononuclear cells to endothelial cell differentiation.

KEYWORDS:

DNA methylation; Differentiation; Endothelial cells; Gene discovery; MeDIP seq; Promoter CpG islands; RNA seq; Umbilical cord blood derived mononuclear cell

PMID:
28887159
DOI:
10.1016/j.gene.2017.09.006
[Indexed for MEDLINE]
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