Epigenetic reprogramming converts human Wharton’s jelly mesenchymal stem cells into functional cardiomyocytes by differential regulation of Wnt mediators

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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0638-7) contains supplementary material, which is available to authorized users.

RNA-Based Strategies for Cardiac Reprogramming of Human Mesenchymal Stromal Cells

“…Yet, it was shown that cytokines like BMP4, IL and TGF improve cardiac development of human and non-human MSCs [57,77]. However, the cardiomyocyte-like cells derived from these programmed MSCs lack profound sarcomere formation, beating activity and physiological maturation [78,79]. This is in accordance to our data indicating that mRNA transfection could promote the expression of early cardiac proteins, while differentiation efficiency and elaboration of a terminal cardiac phenotype is profoundly limited when compared to PSC differentiation protocols [27,31].…”

Small‐molecule–based generation of functional cardiomyocytes from human umbilical cord–derived induced pluripotent stem cells

“…Although the majority of reprogramming studies have focused on fibroblasts as the somatic cell source, several studies have recently demonstrated the feasibility of reprogramming MSCs. 4,13,14 Human umbilical cord tissue is readily available, easily procured without invasive procedures, and does not elicit ethical debate. Cai et al 3 reported the generation of iPSCs from UC-MSCs.…”

Stem cells and physical energies: can we really drive stem cell fate?

“…In the last two centuries, with the development of the electromagnetic theory, there has been a surge increase in the interest to the interaction of electromagnetic fields (EMFs) and various life processes. Particularly into the 20th century, plenty of bio-effects of electric, magnetic, and electromagnetic fields on human beings, animals and cells have been described either in vitro or in vivo conditions (Berg 1997, Berg 1999, Hönes Vol. 68 et al 1998.…”

“…In vitro stem cell differentiation can be induced by chemical stimuli, such as small molecules that act modifying self-renewal, inducing cell differentiation and reprogramming (Ding and Schultz 2004, Mansour et al 2012, Mattout et al 2011. Stem cell fate is controlled by multiple signaling pathways, involving miRNAs (Gangaraju andLin 2009, Balzano et al2018), and other epigenetic events, such as DNA methylation and chromatin remodeling, still not completely yet understood (Lacagnina 2019, Bhuvanalakshmi et al 2017, He et al 2019. Growth factors, hormones, cytokines and other pharmacological agents can be added to the culturing medium to influence stem cell commitment toward specific phenotype (Christ et al 2013, Maioli et al 2016a.…”