DNA Methylation Inhibitors, 5‐azacytidine and Zebularine Potentiate the Transdifferentiation of Rat Bone Marrow Mesenchymal Stem Cells into Cardiomyocytes

Abstract

SUMMARYBackground: Mesenchymal stem cells (MSCs) have immense self-renewal capability. They can be differentiated into many cell types and therefore hold great potential in the field of regenerative medicine. MSCs can be converted into beating cardiomyocytes by treating them with DNA-demethylating agents. Some of these compounds are nucleoside analogs that are widely used for studying the role of DNA methylation in biological processes as well as for the clinical treatment of leukemia and other carcinomas. Aims: To achieve a better therapeutic option for cardiovascular regeneration, this study was carried out using MSCs treated with two synthetic compounds, zebularine and 5-azacytidine. It can be expected that treated MSCs prior to transplantation may increase the likelihood of successful regeneration of damaged myocardium. Methods: The optimized concentrations of these compounds were added separately into the culture medium and the treated cells were analyzed for the expression of cardiac-specific genes by RT-PCR and cardiac-specific proteins by immunocytochemistry and flow cytometry. Treated MSCs were cocultured with cardiomyocytes to see the fusion capability of these cells. Results: mRNA and protein expressions of GATA4, Nkx2.5, and cardiac troponin T were observed in the treated MSCs. Coculture studies of MSCs and cardiomyocytes have shown improved fusion with zebularine-treated MSCs as compared to untreated and 5-azacytidine-treated MSCs. Conclusion: The study is expected to put forth another valuable aspect of certain compounds, that is, induction of transdifferentiation of MSCs into cardiomyocytes. This would serve as a tool for modified cellular therapy and may increase the probability of better myocardial regeneration.

Combination of retinoic acid, dimethyl sulfoxide and 5-azacytidine promotes cardiac differentiation of human fetal liver-derived mesenchymal stem cells

“…Several studies have shown that 5-azacytidine (5-aza), a DNA demethylating chemical compound, induced MSCs to differentiate into cardiomyocytes, while the differentiation ratio is very low (Qian et al 2012;Naeem et al 2013). Retinoic acid (RA) and dimethyl sulfoxide (DMSO) are none specific inducers and they have been found to exert beneficial effect on the induction of cardiogenic differentiation (Lengerke et al 2011;Seya et al 2012), so we hypothesized that the cardiogenic differentiation efficiency of MSCs could be enhanced by the combined treatment of 5-aza, RA and DMSO.…”

Epigenetic mechanisms in heart development and disease

“…In cardiovascular research, 5-azaC has received increased attention as a result of findings that reported the DNA methylation inhibitor promotes the differentiation of cardiomyocytes from mesenchymal stem cells [142–144], although at low frequencies [145]. However, the potential benefits of 5-azaC in cardiac regeneration studies are limited by toxicity, including myeloid suppression and induction of cardiomyocyte cell death.…”

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

“…Other small-molecule supplements, such as 5-azacytidine, alone or in combination, may also increase the yield of cardiomyocytes from pluripotent cell lines [165][166][167]. Studies on murine embryonic carcinoma cells showed that treatment with 5-azacytidine resulted in an increase in the levels of acetylated histone H3; upregulation of the expression of cardio-specific gene markers and in development of cardiomyocyte-rich embryoid bodies [168].…”