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A HCN4+ cardiomyogenic progenitor derived from the first heart field and human pluripotent stem cells

Nature Cell Biology volume 15, pages 10981106 (2013) | Download Citation

Abstract

Most of the mammalian heart is formed from mesodermal progenitors in the first and second heart fields (FHF and SHF), whereby the FHF gives rise to the left ventricle and parts of the atria and the SHF to the right ventricle, outflow tract and parts of the atria1,2,3. Whereas SHF progenitors have been characterized in detail, using specific molecular markers4,5,6,7,8, comprehensive studies on the FHF have been hampered by the lack of exclusive markers. Here, we present Hcn4 (hyperpolarization-activated cyclic nucleotide-gated channel 4) as an FHF marker. Lineage-traced Hcn4+/FHF cells delineate FHF-derived structures in the heart and primarily contribute to cardiomyogenic cell lineages, thereby identifying an early cardiomyogenic progenitor pool. As a surface marker, HCN4 also allowed the isolation of cardiomyogenic Hcn4+/FHF progenitors from human embryonic stem cells. We conclude that a primary purpose of the FHF is to generate cardiac muscle and support the contractile activity of the primitive heart tube, whereas SHF-derived progenitors contribute to heart cell lineage diversification.

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References

  1. 1.

    & How to make a heart: the origin and regulation of cardiac progenitor cells. Curr. Top. Dev. Biol. 90, 1–41 (2010).

  2. 2.

    , , , & Islet1 cardiovascular progenitors: a single source for heart lineages? Development 135, 193–205 (2008).

  3. 3.

    , & Lives of a heart cell: tracing the origins of cardiac progenitors. Cell Stem Cell 2, 320–331 (2008).

  4. 4.

    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).

  5. 5.

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

  6. 6.

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

  7. 7.

    et al. Human ISL1 heart progenitors generate diverse multipotent cardiovascular cell lineages. Nature 460, 113–117 (2009).

  8. 8.

    et al. Islet 1 is expressed in distinct cardiovascular lineages, including pacemaker and coronary vascular cells. Dev. Biol. 304, 286–296 (2007).

  9. 9.

    et al. Epicardial progenitors contribute to the cardiomyocyte lineage in the developing heart. Nature 454, 109–113 (2008).

  10. 10.

    et al. A myocardial lineage derives from Tbx18 epicardial cells. Nature 454, 104–108 (2008).

  11. 11.

    et al. Distinct compartments of the proepicardial organ give rise to coronary vascular endothelial cells. Dev. Cell 22, 639–650 (2012).

  12. 12.

    , , , & Fate of the mammalian cardiac neural crest. Development 127, 1607–1616 (2000).

  13. 13.

    et al. Generation of functional ventricular heart muscle from mouse ventricular progenitor cells. Science 326, 426–429 (2009).

  14. 14.

    & Congenitally corrected transposition of the great arteries. Heart 96, 1154–1161 (2010).

  15. 15.

    et al. Latest insights in therapeutic options for systemic right ventricular failure: a comparison with left ventricular failure. Heart 95, 960–963 (2009).

  16. 16.

    , & Novel insights into the distribution of cardiac HCN channels: an expression study in the mouse heart. J. Mol. Cell. Cardiol. 51, 997–1006 (2011).

  17. 17.

    et al. Funny current channel HCN4 delineates the developing cardiac conduction system in chicken heart. Heart Rhythm 8, 1254–1263 (2011).

  18. 18.

    , & Expression of the hyperpolarization-activated cyclic nucleotide-gated cation channel HCN4 during mouse heart development. Gene Exp. 3, 777–783 (2003).

  19. 19.

    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).

  20. 20.

    et al. Chamber-specific cardiac expression of Tbx5 and heart defects in Holt-Oram syndrome. Dev. Biol. 211, 100–108 (1999).

  21. 21.

    et al. Tamoxifen-inducible gene deletion in the cardiac conduction system. J. Mol. Cell. Cardiol. 45, 62–69 (2008).

  22. 22.

    et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010).

  23. 23.

    , , , & The right ventricle, outflow tract, and ventricular septum comprise a restricted expression domain within the secondary/anterior heart field. Dev. Biol. 287, 134–145 (2005).

  24. 24.

    et al. The renewal and differentiation of Isl1+ cardiovascularprogenitors are controlled by a Wnt/beta-catenin pathway. Cell Stem Cell 1, 165–179 (2007).

  25. 25.

    Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat. Genet. 21, 70–71 (1999).

  26. 26.

    , & Early mesodermal cues assign avian cardiac pacemaker fate potential in a tertiary heart field. Science 428, 821–827 (2013).

  27. 27.

    et al. The sinus venosus progenitors separate and diversify from the first and second heart fields early in development. Cardiovasc. Res. 87, 92–101 (2010).

  28. 28.

    et al. Biphasic development of the mammalian ventricular conduction system. Circ. Res. 107, 153–161 (2010).

  29. 29.

    et al. Cardiomyocytes derived from embryonic stem cells resemble cardiomyocytes of the embryonic heart tube. Cardiovasc. Res. 58, 399–409 (2003).

  30. 30.

    et al. The heart endocardium is derived from vascular endothelial progenitors. Development 138, 4777–4787 (2011).

  31. 31.

    et al. Driving vascular endothelial cell fate of human multipotent Isl1+ heart progenitors with VEGF modified mRNA. Cell Res. (2013).

  32. 32.

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

  33. 33.

    , & Reassessment of Isl1 and Nkx2-5 cardiac fate maps using a Gata4-based reporter of Cre activity. Dev. Biol. 323, 98–104 (2008).

  34. 34.

    et al. Tbx5-hedgehog molecular networks are essential in the second heart field for atrial septation. Dev. Cell 23, 280–291 (2012).

  35. 35.

    & The formation of the embryonic mouse heart: heart fields and myocardial cell lineages. Ann. NY Acad. Sci. 1188, 15–24 (2010).

  36. 36.

    et al. The Tbx2 primary myocardium of the atrioventricular canal forms the atrioventricular node and the base of the left ventricle. Circ. Res. 104, 1267–1274 (2009).

  37. 37.

    & Tracing cells for tracking cell lineage and clonal behavior. Dev. Cell 21, 394–409 (2011).

  38. 38.

    , & A somitic compartment of tendon progenitors. Cell 113, 235–248 (2003).

  39. 39.

    , & A control of dorsoventral pattern in the chick paraxial mesoderm. Development 124, 3895–3908 (1997).

  40. 40.

    et al. Wnt/β-catenin signaling in the dental mesenchymeregulates incisor development by regulating Bmp4. Dev. Biol. 348, 97–106 (2010).

  41. 41.

    & Histochemical methods for separate, consecutive and simultaneous demonstration of acetylcholinesterase and norepinephrine in cryostat sections. J. Histochem. Cytochem. 15, 580–588 (1967).

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Acknowledgements

We would like to thank C. Cowan for his support, L. Prickett-Rice, K. Folz-Donahue and M. Weglarz of the Harvard Stem Cell Institute Flow Cytometry Core Facility for assistance with FACS analysis, C. Du of the Tufts Electrophysiology Core for assistance with electrophysiology recordings, L. Bu for technical advice, K. Buac, E. Hansson, C. Riedel and L. Bu for critical reading of the manuscript and discussions, and C. Hartmann for advice on double-fluorescence in situ hybridizations. D.S. has received a D.F.G. (German Research Foundation) postdoctoral fellowship. K.B. was supported by a NHLBI T32HL007208 grant. This work is financially supported by the NIH U01 HL098 166 and NIH U01H100408 research grants.

Author information

Author notes

    • Monika K. Abramczuk
    • , Kristina Buac
    •  & Lior Zangi

    Present addresses: Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), Dr Bohr-Gasse 3, 1030 Vienna, Austria (M.K.A.); Department of Genetics, University of Georgia, Athens, Georgia 30602, USA (K.B.); Department of Cardiology, Children’s Hospital Boston, Boston 02115, USA, and Harvard Stem Cell Institute, Cambridge, Massachusetts, USA (L.Z.)

Affiliations

  1. Cardiovascular Research Center, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA

    • Daniela Später
    • , Monika K. Abramczuk
    • , Kristina Buac
    • , Lior Zangi
    • , Maxine W. Stachel
    • , Jonathan Clarke
    • , Makoto Sahara
    •  & Kenneth R. Chien
  2. Department of Stem Cell and Regenerative Biology, Harvard University and Harvard Medical School, 7 Divinity Avenue, Cambridge, Massachusetts 02138, USA

    • Daniela Später
    • , Monika K. Abramczuk
    • , Kristina Buac
    • , Lior Zangi
    • , Maxine W. Stachel
    • , Makoto Sahara
    •  & Kenneth R. Chien
  3. Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Friedrich-Alexander-Universität, Erlangen-Nürnberg, 91054 Erlangen, Germany

    • Andreas Ludwig
  4. Department of Cell and Molecular Biology and Medicine, Karolinska Institutet, 7 Berzelius Vag, Stockholm, Sweden

    • Kenneth R. Chien

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Contributions

D.S. and K.R.C. designed the experiments and wrote the manuscript. D.S. carried out most of the experiments and analysed most of the data. M.K.A. helped to perform and analyse clonal analysis experiments. K.B. contributed PCRs with reverse transcription of in vivo clonal analysis. L.Z. performed some immunohistochemical staining, M.W.S. helped with some hESC differentiation assays and RNA isolations, J.C. performed acetylcholinesterase staining, M.S. helped with some mESC clonal analysis cultures and A.L. provided the Hcn4CreErt2 mouse line.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Kenneth R. Chien.

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    Clonal analysis of single Hcn4+/FHF cells isolated from mouse embryos.

    Representative example of a beating cardiomyogenic clone derived from a single Hcn4+/FHF cell from mouse embryos.

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DOI

https://doi.org/10.1038/ncb2824

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