TY - JOUR
T1 - Induction of diverse cardiac cell types by reprogramming fibroblasts with cardiac transcription factors
AU - Nam, Young Jae
AU - Lubczyk, Christina
AU - Bhakta, Minoti
AU - Zang, Tong
AU - Fernandez-Perez, Antonio
AU - McAnally, John
AU - Bassel-Duby, Rhonda
AU - Olson, Eric N.
AU - Munshi, Nikhil V.
N1 - Publisher Copyright:
© 2014, Published by The Company of Biologists Ltd.
PY - 2014/11/1
Y1 - 2014/11/1
N2 - 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.
AB - 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.
KW - Cardiomyocyte
KW - Direct reprogramming
KW - Mouse
KW - Pacemaker
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U2 - 10.1242/dev.114025
DO - 10.1242/dev.114025
M3 - Article
C2 - 25344074
AN - SCOPUS:84908431264
SN - 0950-1991
VL - 141
SP - 4267
EP - 4278
JO - Development (Cambridge)
JF - Development (Cambridge)
IS - 22
ER -