Induction of diverse cardiac cell types by reprogramming fibroblasts with cardiac transcription factors

Young Jae Nam, Christina Lubczyk, Minoti Bhakta, Tong Zang, Antonio Fernandez-Perez, John McAnally, Rhonda Bassel-Duby, Eric N. Olson, Nikhil V. Munshi

Research output: Contribution to journalArticlepeer-review

75 Scopus citations

Abstract

Various combinations of cardiogenic transcription factors, including Gata4 (G), Hand2 (H), Mef2c (M) and Tbx5 (T), can reprogram fibroblasts into induced cardiac-like myocytes (iCLMs) in vitro and in vivo. Given that optimal cardiac function relies on distinct yet functionally interconnected atrial, ventricular and pacemaker (PM) cardiomyocytes (CMs), it remains to be seen which subtypes are generated by direct reprogramming and whether this process can be harnessed to produce a specific CM of interest. Here, we employ a PM-specificHcn4-GFPreportermouse and a spectrumofCMsubtypespecific markers to investigate the range of cellular phenotypes generated by reprogramming of primary fibroblasts. Unexpectedly, we find that a combination of four transcription factors (4F) optimized for Hcn4-GFP expression does not generate beating PM cells due to inadequate sarcomeric protein expression and organization. However, applying strict single-cell criteria to GHMT-reprogrammed cells, we observe induction of diverse cellular phenotypes, including those resembling immature forms of all three major cardiac subtypes (i.e. atrial, ventricular and pacemaker). In addition, we demonstrate that cells induced by GHMT are directly reprogrammed and do not arise from an Nxk2.5+ progenitor cell intermediate. Taken together, our results suggest a remarkable degree of plasticity inherent to GHMT reprogramming and provide a starting point for optimization of CM subtype-specific reprogramming protocols.

Original languageEnglish (US)
Pages (from-to)4267-4278
Number of pages12
JournalDevelopment (Cambridge)
Volume141
Issue number22
DOIs
StatePublished - Nov 1 2014

Keywords

  • Cardiomyocyte
  • Direct reprogramming
  • Mouse
  • Pacemaker

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology

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