Abstract
BackgroundLineage commitment of mesenchymal stem cells (MSCs) to cardiac differentiation is controlled by transcription factors that are regulated by epigenetic events, mainly histone deacetylation and promoter DNA methylation. Here, we studied the differentiation of human Wharton’s jelly MSCs (WJMSCs) into the cardiomyocyte lineage via epigenetic manipulations.MethodsWe introduced these changes using inhibitors of DNA methyl transferase and histone deacetylase, DC301, DC302, and DC303, in various combinations. We characterized for cardiogenic differentiation by assessing the expression of cardiac-specific markers by immunolocalization, quantitative RT-PCR, and flow cytometry. Cardiac functional studies were performed by FURA2AM staining and Greiss assay. The role of Wnt signaling during cardiac differentiation was analyzed by quantitative RT-PCR. In-vivo studies were performed in a doxorubicin-induced cardiotoxic mouse model by injecting cardiac progenitor cells. Promoter methylation status of the cardiac transcription factor Nkx2.5 and the Wnt antagonist, secreted frizzled-related protein 4 (sFRP4), after cardiac differentiation was studied by bisulfite sequencing.ResultsBy induction with DC301 and DC302, WJMSCs differentiated into cardiomyocyte-like structures with an upregulation of Wnt antagonists, sFRP3 and sFRP4, and Dickkopf (Dkk)1 and Dkk3. The cardiac function enhancer, vinculin, and DDX20, a DEAD-box RNA helicase, were also upregulated in differentiated cardiomyocytes. Additionally, bisulfite sequencing revealed, for the first time in cardiogenesis, that sFRP4 is activated by promoter CpG island demethylation. In vivo, these MSC-derived cardiac progenitors could not only successfully engraft to the site of cardiac injury in mice with doxorubicin-induced cardiac injury, but also form functional cardiomyocytes and restore cardiac function.ConclusionThe present study unveils a link between Wnt inhibition and epigenetic modification to initiate cardiac differentiation, which could enhance the efficacy of stem cell therapy for ischemic heart disorders.
Highlights
Lineage commitment of mesenchymal stem cells (MSCs) to cardiac differentiation is controlled by transcription factors that are regulated by epigenetic events, mainly histone deacetylation and promoter DNA methylation
We characterized for the gene expression of pluripotency markers such as Sox2, Oct4, and Nanog, and for the MSC positive CD marker CD44 and negative marker CD34, and found that Oct 4, Nanog, and CD44 were highly expressed whereas CD34 expression was minimal (Additional file 2: Figure S1A3)
We analyzed the expression of early (Nkx2.5, GATA4) and late (MLC, troponin T (TnT), cardiac actin) cardiac markers in MSCs differentiated by various combinations of epigenetic modifiers using quantitative RT-PCR (Fig. 1b)
Summary
Lineage commitment of mesenchymal stem cells (MSCs) to cardiac differentiation is controlled by transcription factors that are regulated by epigenetic events, mainly histone deacetylation and promoter DNA methylation. MSCs from various fetal and adult tissues are excellent viable options for stem cell therapy for cardiac degeneration. During myocardial damage and injury, cardiac progenitor cells migrate to the site of injury to augment the limited mitotic capacity of terminally differentiated cardiomyocytes and eventually regenerate the myocardium [4]. These cell populations are very few in number and decrease significantly during the aging process, thereby compromising the regenerative process [5]. MSCs from adult tissues differ from MSCs from tissues of fetal origin by way of their proliferative ability, immunosuppressive properties, and therapeutic efficacy; the use of Wharton’s jelly, the primitive connective tissue of the human umbilical cord, is a viable alternate perinatal source of MSCs [8, 9]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
