Abstract
Regulation of multipotent cardiac progenitor cell (CPC) expansion and subsequent differentiation into cardiomyocytes, smooth muscle or endothelial cells is a fundamental aspect of basic cardiovascular biology and cardiac regenerative medicine. However, the mechanisms governing these decisions remain unclear. Here, we show that Wnt/β-catenin signalling, which promotes expansion of CPCs1,2,3, is negatively regulated by Notch1-mediated control of phosphorylated β-catenin accumulation within CPCs, and that Notch1 activity in CPCs is required for their differentiation. Notch1 positively, and β-catenin negatively, regulated expression of the cardiac transcription factors, Isl1, Myocd and Smyd1. Surprisingly, disruption of Isl1, normally expressed transiently in CPCs before their differentiation4, resulted in expansion of CPCs in vivo and in an embryonic stem (ES) cell system. Furthermore, Isl1 was required for CPC differentiation into cardiomyocyte and smooth muscle cells, but not endothelial cells. These findings reveal a regulatory network controlling CPC expansion and cell fate that involves unanticipated functions of β-catenin, Notch1 and Isl1 that may be leveraged for regenerative approaches involving CPCs.
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Acknowledgements
We thank R. Kopan (Washington Univeristy, St. Louis, MO) and M. M. Taketo (Kyoto University, Kyoto, Japan) for providing Notch1flox and β-catenin/loxP(ex3)loxP mice, respectively. The authors thank G. Howard and S. Ordway for editorial assistance, R.F. Yeh for statistical analyses, K. Cordes for graphical assistance, B. Taylor for manuscript and figure preparation and Srivastava lab members for helpful discussions. C.K. was supported by a fellowship from the American Heart Association (AHA) and California Institute for Regenerative Medicine (CIRM); D.S. was an Established Investigator of the AHA and was supported by grants from NHLBI/NIH and CIRM. This work was also supported by NIH/NCRR grant (C06 RR018928) to the Gladstone Institutes.
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C.K. designed, performed, supervised in vivo and in vitro work and wrote the manuscript; L.Q. performed flow cytometry and EMSA, and contributed in luciferase assays; P.C. designed and performed Isl1 gain-of-function studies and contributed in ChIP and luciferase assays; V.N. performed β-catenin western and Top/Fop flash assays; J.A. contributed to ChIP assays; D.S. designed and supervised this work and wrote the manuscript.
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P.S. serves on the scientific advisory board of iPierian.
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Kwon, C., Qian, L., Cheng, P. et al. A regulatory pathway involving Notch1/β-catenin/Isl1 determines cardiac progenitor cell fate.. Nat Cell Biol 11, 951–957 (2009). https://doi.org/10.1038/ncb1906
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DOI: https://doi.org/10.1038/ncb1906
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