DNA methylation microarray uncovers a permissive methylome for cardiomyocyte differentiation in human mesenchymal stem cells

Kavitha Govarthanan; Piyush Kumar Gupta; Deepa Ramasamy; Pavitra Kumar; Shobana Mahadevan; Rama Shanker Verma

doi:10.1016/j.ygeno.2019.08.007

DNA methylation microarray uncovers a permissive methylome for cardiomyocyte differentiation in human mesenchymal stem cells

Genomics. 2020 Mar;112(2):1384-1395. doi: 10.1016/j.ygeno.2019.08.007. Epub 2019 Aug 12.

Authors

Kavitha Govarthanan¹, Piyush Kumar Gupta¹, Deepa Ramasamy¹, Pavitra Kumar², Shobana Mahadevan³, Rama Shanker Verma⁴

Affiliations

¹ Stem cells and Molecular Biology Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, Tamilnadu, India.
² AU-KBC Research Centre, Madras Institute of Technology, Anna University, Chennai 600044, Tamilnadu, India.
³ Seethapathy Clinic and Hospital, Royapettah, Chennai 60014, Tamilnadu, India.
⁴ Stem cells and Molecular Biology Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, Tamilnadu, India. Electronic address: vermars@iitm.ac.in.

PMID: 31415810
DOI: 10.1016/j.ygeno.2019.08.007

Abstract

Differentiation of Wharton's Jelly-Mesenchymal Stem cells (WJ-MSCs) into cardiomyocytes (CMs) in vitro has been reported widely although contradictions remain regarding the maturation of differentiated MSCs into fully functioning CMs. Studies suggest that use of epigenetic modifiers like 5'Azacytidine (5-AC) in MSCs de-methylates DNA and results in expression of cardiac-specific genes (CSGs). However, only partial expression of the CSG set leads to incomplete differentiation of WJ-MSCs to CMs. We used the Agilent 180 K human DNA methylation microarray on WJ-MSCs, 5-AC treated WJ-MSCs and human cardiac tissue (hCT) to analyze differential DNA methylation profiles which were then validated by bisulfite sequencing PCR (BSP). BSP confirmed that only a limited number of CSGs were de-methylated by 5-AC in WJ-MSCs. It also revealed that hCT displays a methylation profile similar to promoter regions of CSG in untreated WJ-MSCs. Thus, the presence of hypo-methylated CSGs indicates that WJ-MSCs are ideal cell types for cardiomyogenic differentiation.

Keywords: 5-Azacytidine; CpG; DNA methylation; DNA microarray; Differentiation; Mesenchymal stem cells; Promoter.

Publication types

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

MeSH terms

Cell Differentiation*
Cells, Cultured
DNA Methylation*
Epigenome*
Fetal Blood / cytology
Humans
Mesenchymal Stem Cells / cytology
Mesenchymal Stem Cells / metabolism*
Myocytes, Cardiac / cytology
Myocytes, Cardiac / metabolism*