Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

A regulatory pathway involving Notch1/β-catenin/Isl1 determines cardiac progenitor cell fate.

Nature Cell Biology volume 11, pages 951–957 (2009)Cite this article

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Notch1 loss-of-function causes CPC expansion and increases free β-catenin levels.
Figure 2: Identification of genes affected by stabilized β-catenin in cardiac progenitors.
Figure 3: Isl1 loss-of-function results in expansion of CPCs and suppression of their myocardial and smooth muscle lineages.
Figure 4: Increased levels of Isl1 promote myocardial differentiation.
Figure 5: Isl1 targets Myocd and β-catenin regulates Bhlhb2 to repress Smyd1.

Similar content being viewed by others

Accession codes

Accessions

Gene Expression Omnibus

References

  1. Cohen, E. D. et al. Wnt/β-catenin signaling promotes expansion of Isl-1-positive cardiac progenitor cells through regulation of FGF signaling. J. Clin. Invest. 117, 1794–1804 (2007).

    Article  CAS  Google Scholar 

  2. Kwon, C. et al. Canonical Wnt signaling is a positive regulator of mammalian cardiac progenitors. Proc. Natl Acad. Sci. USA 104, 10894–10899 (2007).

    Article  CAS  Google Scholar 

  3. Qyang, Y. et al. The renewal and differentiation of Isl1+ cardiovascular progenitors are controlled by a Wnt/β-catenin pathway. Cell Stem Cell 1, 165–179 (2007).

    Article  CAS  Google Scholar 

  4. Cai, C. L. et al. Isl1 identifies a cardiac progenitor population that proliferates prior to differentiation and contributes a majority of cells to the heart. Dev. Cell 5, 877–889 (2003).

    Article  CAS  Google Scholar 

  5. Yang, L. et al. Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell-derived population. Nature 453, 524–528 (2008).

    Article  CAS  Google Scholar 

  6. Buckingham, M., Meilhac, S. & Zaffran, S. Building the mammalian heart from two sources of myocardial cells. Nature Rev. Genet. 6, 826–835 (2005).

    Article  CAS  Google Scholar 

  7. Wu, S. M. et al. Developmental origin of a bipotential myocardial and smooth muscle cell precursor in the mammalian heart. Cell 127, 1137–1150 (2006).

    Article  CAS  Google Scholar 

  8. Moretti, A. et al. Multipotent embryonic isl1+ progenitor cells lead to cardiac, smooth muscle, and endothelial cell diversification. Cell 127, 1151–1165 (2006).

    Article  CAS  Google Scholar 

  9. Laugwitz, K. L. et al. Postnatal isl1+ cardioblasts enter fully differentiated cardiomyocyte lineages. Nature 433, 647–653 (2005).

    Article  CAS  Google Scholar 

  10. Kattman, S. J., Huber, T. L. & Keller, G. M. Multipotent flk-1+ cardiovascular progenitor cells give rise to the cardiomyocyte, endothelial, and vascular smooth muscle lineages. Dev. Cell 11, 723–732 (2006).

    Article  CAS  Google Scholar 

  11. Srivastava, D. Making or breaking the heart: from lineage determination to morphogenesis. Cell 126, 1037–1048 (2006).

    Article  CAS  Google Scholar 

  12. Christoforou, N. et al. Mouse ES cell-derived cardiac precursor cells are multipotent and facilitate identification of novel cardiac genes. J. Clin. Invest. 118, 894–903 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Lyons, I. et al. Myogenic and morphogenetic defects in the heart tubes of murine embryos lacking the homeobox gene Nkx2–5 Genes Dev. 9, 1654–1666 (1995).

    Article  CAS  Google Scholar 

  14. Stanley, E. G. et al. Efficient Cre-mediated deletion in cardiac progenitor cells conferred by a 3´UTR-ires-Cre allele of the homeobox gene Nkx2–5 Int. J. Dev. Biol. 46, 431–439 (2002).

    CAS  PubMed  Google Scholar 

  15. Prall, O. W. et al. An Nkx2–5/Bmp2/Smad1 negative feedback loop controls heart progenitor specification and proliferation. Cell 128, 947–959 (2007).

    Article  CAS  Google Scholar 

  16. Hayward, P., Kalmar, T. & Arias, A. M. Wnt/Notch signalling and information processing during development. Development 135, 411–424 (2008).

    Article  CAS  Google Scholar 

  17. Rones, M. S., McLaughlin, K. A., Raffin, M. & Mercola, M. Serrate and Notch specify cell fates in the heart field by suppressing cardiomyogenesis. Development 127, 3865–3876 (2000).

    CAS  PubMed  Google Scholar 

  18. Nemir, M., Croquelois, A., Pedrazzini, T. & Radtke, F. Induction of cardiogenesis in embryonic stem cells via downregulation of Notch1 signaling. Circ. Res. 98, 1471–1478 (2006).

    Article  CAS  Google Scholar 

  19. Radtke, F. et al. Deficient T cell fate specification in mice with an induced inactivation of Notch1. Immunity 10, 547–558 (1999).

    Article  CAS  Google Scholar 

  20. Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001).

    Article  CAS  Google Scholar 

  21. Hsiao, E. C. et al. Marking embryonic stem cells with a 2A self-cleaving peptide: a NKX2–5 emerald GFP BAC reporter. PLoS ONE 3, e2532 (2008).

    Article  Google Scholar 

  22. Votin, V., Nelson, W. J. & Barth, A. I. Neurite outgrowth involves adenomatous polyposis coli protein and beta-catenin. J. Cell Sci. 118, 5699–5708 (2005).

    Article  CAS  Google Scholar 

  23. Gottlieb, P. D. et al. Bop encodes a muscle-restricted protein containing MYND and SET domains and is essential for cardiac differentiation and morphogenesis. Nature Genet. 31, 25–32 (2002).

    Article  CAS  Google Scholar 

  24. Li, S., Wang, D. Z., Wang, Z., Richardson, J. A. & Olson, E. N. The serum response factor coactivator myocardin is required for vascular smooth muscle development. Proc. Natl Acad. Sci. USA 100, 9366–9370 (2003).

    Article  CAS  Google Scholar 

  25. Tan, X., Rotllant, J., Li, H., De Deyne, P. & Du, S. J. SmyD1, a histone methyltransferase, is required for myofibril organization and muscle contraction in zebrafish embryos. Proc. Natl Acad. Sci. USA 103, 2713–2718 (2006).

    Article  CAS  Google Scholar 

  26. Ueyama, T., Kasahara, H., Ishiwata, T., Nie, Q. & Izumo, S. Myocardin expression is regulated by Nkx2.5, and its function is required for cardiomyogenesis. Mol. Cell Biol. 23, 9222–9232 (2003).

    Article  CAS  Google Scholar 

  27. Wang, D. et al. Activation of cardiac gene expression by myocardin, a transcriptional cofactor for serum response factor. Cell 105, 851–862 (2001).

    Article  CAS  Google Scholar 

  28. Zawel, L. et al. DEC1 is a downstream target of TGF-β with sequence-specific transcriptional repressor activities. Proc. Natl Acad. Sci. USA 99, 2848–2853 (2002).

    Article  CAS  Google Scholar 

  29. Shen, M. et al. Basic helix-loop-helix protein DEC1 promotes chondrocyte differentiation at the early and terminal stages. J. Biol. Chem. 277, 50112–50120 (2002).

    Article  CAS  Google Scholar 

  30. Honma, S. et al. Dec1 and Dec2 are regulators of the mammalian molecular clock. Nature 419, 841–844 (2002).

    Article  CAS  Google Scholar 

  31. Harada, N. et al. Intestinal polyposis in mice with a dominant stable mutation of the β-catenin gene. EMBO J. 18, 5931–5942 (1999).

    Article  CAS  Google Scholar 

  32. Kwon, C., Hays, R., Fetting, J. & Orenic, T. V. Opposing inputs by Hedgehog and Brinker define a stripe of hairy expression in the Drosophila leg imaginal disc. Development 131, 2681–2692 (2004).

    Article  CAS  Google Scholar 

  33. Kwon, C., Han, Z., Olson, E. N. & Srivastava, D. MicroRNA1 influences cardiac differentiation in Drosophila and regulates Notch signaling. Proc. Natl Acad. Sci. USA 102, 18986–18991 (2005).

    Article  CAS  Google Scholar 

  34. Dodou, E., Verzi, M. P., Anderson, J. P., Xu, S. M. & Black, B. L. Mef2c is a direct transcriptional target of ISL1 and GATA factors in the anterior heart field during mouse embryonic development. Development 131, 3931–3942 (2004).

    Article  CAS  Google Scholar 

Download references

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.

Author information

Author notes

  1. Chulan Kwon, Li Qian, Paul Cheng, Vishal Nigam, Joshua Arnold and Deepak Srivastava: These authors contributed equally in this work.

Authors and Affiliations

  1. Gladstone Institute of Cardiovascular Disease and Departments of Pediatrics and Biochemistry & Biophysics, University of California, San Francisco, 1650 Owens Street, San Francisco, 94158, CA, USA

    Chulan Kwon, Li Qian, Paul Cheng, Vishal Nigam, Vishal Nigam, Joshua Arnold & Deepak Srivastava

Authors
  1. Chulan Kwon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Li Qian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paul Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vishal Nigam
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Joshua Arnold
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Deepak Srivastava
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

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.

Corresponding authors

Correspondence to Chulan Kwon or Deepak Srivastava.

Ethics declarations

Competing interests

P.S. serves on the scientific advisory board of iPierian.

Supplementary information

Supplementary Information

Supplementary Information (PDF 606 kb)

Supplementary Information

Supplementary Table 1 (XLS 62 kb)

Supplementary Information

Supplementary Table 2 (XLS 37 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1906

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing