Cardiomyogenic differentiation of human adipose‐derived mesenchymal stem cells transduced with Tbx20‐encoding lentiviral vectors

Abstract

Ischemic heart disease often results in myocardial infarction and is the leading cause of mortality and morbidity worldwide. Improvement in the function of infarcted myocardium is a main purpose of cardiac regenerative medicine. One possible way to reach this goal is via stem cell therapy. Mesenchymal stem cells (MSCs) are multipotent stromal cells that can differentiate into a variety of cell types but display limited cardiomyogenic differentiation potential. Members of the T-box family of transcription factors including Tbx20 play important roles in heart development and cardiomyocyte homeostasis. Therefore, in the current study, we investigated the potential of Tbx20 to enhance the cardiomyogenic differentiation of human adipose-derived MSCs (ADMSCs). Human ADMSCs were transduced with a bicistronic lentiviral vector encoding Tbx20 (murine) and the enhanced green fluorescent protein (eGFP) and analyzed 7 and 14 days post transduction. Transduction of human ADMSCs with this lentiviral vector increased the expression of the cardiomyogenic differentiation markers ACTN1, TNNI3, ACTC1, NKX2.5, TBX20 (human), and GATA4 as revealed by RT-qPCR. Consistently, immunocytological results showed elevated expression of α-actinin and cardiac troponin I in these cells in comparison to the cells transduced with control lentiviral particles coding for eGFP alone. Accordingly, forced expression of Tbx20 exerts cardiomyogenic effects on human ADMSCs by increasing the expression of cardiomyogenic differentiation markers at the RNA and protein level.

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

“…However, no clear sarcomere structures have been observed, suggesting a premature cardiac progenitor state. Similarly, forced expression of another factor of the T-box family, TBX20, provokes an up-regulation of sarcomeric proteins, without cardiomyocyte specific sarcomere organization [72]. These data are in line with our observations as we could also detect a moderate signal for α-actinin, albeit without the presence of sarcomere structures (Figure 4).…”

Current Strategies to Enhance Adipose Stem Cell Function: An Update

“…In the case of cardiomyogenic lineage, the forced expression of Tbx20 , a critical transcription factor that contributes to heart development and cardiomyocyte regeneration, efficiently induced expression of cardiomyogenic differentiation markers on ASCs at 14 days after transduction both at the RNA and protein level [135]. It might be necessary to evaluate the cardiomyogenic regenerative capacity of Tbx20-overexpressed ASCs in an ischemic heart disease animal model.…”

β‐Adrenergic stimuli and rotating suspension culture enhance conversion of human adipogenic mesenchymal stem cells into highly conductive cardiac progenitors

“…Recently, Neshati et al () studied the influence of TBX20 on hADMSCs, helping corroborate the importance of the TBX family in the development of CM‐like cells. Their study shows important activation of cardiac transcription factors GATA4, NKX2.5, and actinin, in a similar fashion as our work.…”

“…Many approaches have been taken to overcome this challenge by taking advantage of this principle. In one such study by Okura et al (2010) Recently, Neshati et al (2018) studied the influence of TBX20 on hADMSCs, helping corroborate the importance of the TBX family in the development of CM-like cells. Their study shows important activation of cardiac transcription factors GATA4, NKX2.5, and actinin, in a similar fashion as our work.…”