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Published on: June 3, 2018
Myocyte-specific enhancer factor 2c triggers transdifferentiation of adipose tissue-derived stromal cells into
Shinichiro Takashima1, Soichiro Usui2, Oto Inoue2
1Department of Cardiovascular Medicine, Graduate School of Medical Science, Kanazawa University, 13-1 Takara-machi, Kanazawa, Ishikawa, 920-8641, Japan. takashin@m-kanazawa.jp.
Insights
Adipose tissue stem cells can become beating heart cells. The transcription factor Mef2c is identified as a master gene that drives this cardiac lineage differentiation, paving the way for new heart regeneration therapies.
Area of Science:
- Stem cell biology
- Cardiovascular research
- Molecular genetics
Background:
- Cardiomyocyte regeneration is severely limited in adult mammals.
- Adipose tissue-derived stromal vascular fraction (Ad-SVF) contains stem cells with limited cardiomyocyte transdifferentiation potential.
- The factors regulating Ad-SVF differentiation into cardiomyocytes are largely unknown.
Purpose of the Study:
- To investigate the potential of adult murine Ad-SVF to transdifferentiate into cardiomyocyte-like cells.
- To identify the key molecular regulators of this cardiac lineage differentiation.
- To explore the therapeutic potential of gene-modified Ad-SVF for cardiac regeneration.
Main Methods:
- Development of a simple culture protocol for Ad-SVF transdifferentiation.
- Characterization of spontaneously beating cardiomyocyte-like cells (beating CMs) derived from Ad-SVF.
- Single-cell transcriptome analysis of beating CM-fated progenitors (CFPs).
- Lentiviral transduction of Ad-SVF with Mef2c and gene silencing experiments.
Main Results:
- Adult murine Ad-SVF reproducibly transdifferentiated into beating CMs with ventricular cardiomyocyte phenotype.
- Synchronized intracellular calcium oscillations were observed in beating CMs.
- Myocyte-specific enhancer 2c (Mef2c) was identified as a key transcription factor regulating differentiation.
- Mef2c overexpression enhanced cardiac troponin T expression tenfold, while Mef2c silencing abolished differentiation.
Conclusions:
- A novel culture method enables Ad-SVF to transdifferentiate into functional beating CMs.
- Mef2c is identified as a master regulatory gene for Ad-SVF cardiac lineage commitment.
- These findings support the development of gene-modified Ad-SVF for cardiac regeneration therapies.
Abstract:
Cardiomyocyte regeneration is limited in adults. The adipose tissue-derived stromal vascular fraction (Ad-SVF) contains pluripotent stem cells that rarely transdifferentiate into spontaneously beating cardiomyocyte-like cells (beating CMs). However, the characteristics of beating CMs and the factors that regulate the differentiation of Ad-SVF toward the cardiac lineage are unknown. We developed a simple culture protocol under which the adult murine inguinal Ad-SVF reproducibly transdifferentiates into beating CMs without induction. The beating CMs showed the striated ventricular phenotype of cardiomyocytes and synchronised oscillation of the intracellular calcium concentration among cells on day 28 of Ad-SVF primary culture. We also identified beating CM-fated progenitors (CFPs) and performed single-cell transcriptome analysis of these CFPs. Among 491 transcription factors that were differentially expressed (≥ 1.75-fold) in CFPs and the beating CMs, myocyte-specific enhancer 2c (Mef2c) was key. Transduction of Ad-SVF cells with Mef2c using a lentiviral vector yielded CFPs and beating CMs with ~ tenfold higher cardiac troponin T expression, which was abolished by silencing of Mef2c. Thus, we identified the master gene required for transdifferentiation of Ad-SVF into beating CMs. These findings will facilitate the development of novel cardiac regeneration therapies based on gene-modified, cardiac lineage-directed Ad-SVF cells.
