Article | Published:

ERBB2 triggers mammalian heart regeneration by promoting cardiomyocyte dedifferentiation and proliferation

Nature Cell Biology volume 17, pages 627638 (2015) | Download Citation

Abstract

The murine neonatal heart can regenerate after injury through cardiomyocyte (CM) proliferation, although this capacity markedly diminishes after the first week of life. Neuregulin-1 (NRG1) administration has been proposed as a strategy to promote cardiac regeneration. Here, using loss- and gain-of-function genetic tools, we explore the role of the NRG1 co-receptor ERBB2 in cardiac regeneration. NRG1-induced CM proliferation diminished one week after birth owing to a reduction in ERBB2 expression. CM-specific Erbb2 knockout revealed that ERBB2 is required for CM proliferation at embryonic/neonatal stages. Induction of a constitutively active ERBB2 (caERBB2) in neonatal, juvenile and adult CMs resulted in cardiomegaly, characterized by extensive CM hypertrophy, dedifferentiation and proliferation, differentially mediated by ERK, AKT and GSK3β/β-catenin signalling pathways. Transient induction of caERBB2 following myocardial infarction triggered CM dedifferentiation and proliferation followed by redifferentiation and regeneration. Thus, ERBB2 is both necessary for CM proliferation and sufficient to reactivate postnatal CM proliferative and regenerative potentials.

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Acknowledgements

This work was supported by grants to E.T. from the European Research Council, Israel Science Foundation, the Louis and Fannie Tolz Collaborative Research Project, Estate of Jack Gitlitz and to R.P.H. from the National Health and Medical Research Council (NHMRC) of Australia (573705; 573732) and the Australian Research Council Special Initiative in Stem Cell Science (Stem Cells Australia). We thank C. Birchmeier (MDC, Germany) for providing the Erbb2flox and Erbb2LacZ mice, I. Biton for helping with the MRI measurements, and D. Sawyer (Vanderbilt University, USA), R. Graham (Victor Chang Cardiac Research Institute, Australia), A. Aronheim (Technion, Israel) and K. Yaniv (Weizmann Institute of Science, Israel) for fruitful discussions. We also thank H. Sadek and E. Olson (UT Southwestern Medical Center, USA) for teaching us the neonatal cardiac regenerative model and L. Field (Indiana University School of Medicine, USA) for sharing unpublished data.

Author information

Affiliations

  1. Department of Biological Regulation, Weizmann Institute of Science, Rehovot 76100, Israel

    • Gabriele D’Uva
    • , Alla Aharonov
    • , Mattia Lauriola
    • , Yfat Yahalom-Ronen
    • , Silvia Carvalho
    • , Karen Weisinger
    • , Elad Bassat
    • , Dana Rajchman
    • , Oren Yifa
    • , Marina Lysenko
    • , Michal Neeman
    • , Yosef Yarden
    • , Rachel Sarig
    •  & Eldad Tzahor
  2. Department of Experimental, Diagnostic and Specialty Medicine—DIMES, University of Bologna, 40138 Bologna, Italy

    • Mattia Lauriola
  3. Tamman Cardiovascular Research Institute, Leviev Heart Center, Chaim Sheba Medical Center, Tel Aviv 52621, Israel

    • David Kain
    • , Tal Konfino
    •  & Jonathan Leor
  4. Department of Pediatric Cardiology, Chaim Sheba Medical Center, Tel Aviv 52621, Israel

    • Julius Hegesh
  5. Department of Veterinary Resources, Weizmann Institute of Science, Rehovot 76100, Israel

    • Ori Brenner
  6. Victor Chang Cardiac Research Institute, Darlinghurst 2010, Australia

    • Richard P. Harvey
  7. St Vincent’s Clinical School, University of New South Wales, Kensington 2052, Australia

    • Richard P. Harvey
  8. School of Biological and Biomolecular Sciences, University of New South Wales, Kensington 2052, Australia

    • Richard P. Harvey

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Contributions

G.D’U. and E.T. conceived and designed the experiments. G.D’U. with help from A.A. carried out most of the experiments and analysed the data. M. Lauriola and S.C. performed some western blots and helped with animal studies. D.K. performed myocardial infarction experiment in adult mice. Y.Y-R. performed time-lapse imaging. K.W. and T.K. assisted with myocardial infarction experiment in juvenile mice. E.B., D.R. and O.Y. assisted with gene expression analysis. M. Lysenko performed MRI analysis. O.B. helped with interpretation of histological slides. J.H. helped with the interpretation of echocardiographic analysis. R.S. contributed to the planning and progression of the project. J.L., Y.Y. and M.N. supervised the experiments done by their laboratory members, and E.T. supervised the entire project. R.P.H. provided conceptual inputs and G.D’U., R.P.H. and E.T. wrote the manuscript with editing contributions from all of the authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Eldad Tzahor.

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Videos

  1. 1.

    Cardiac MRI of a control mouse.

    4 chamber cardiac MRI of P22 ctrl mouse.

  2. 2.

    Cardiac MRI of caErbb2 mouse.

    4 chamber cardiac MRI of P22 juvenile caErbb2 mouse generated according to the schema in Fig. 3b.

  3. 3.

    Time-lapse movie of karyokinesis plus cytokinesis (cell division) in mono-nucleated caErbb2 CMs.

    This is a representative time-lapse video of P7 caErbb2 mono-nucleated CMs performing karyokinesis plus cytokinesis (cell division). Heart cells were isolated from P7 caErbb2mice, cultured in vitro for 48 h, then labelled with TMRE to identify CMs (see ‘Methods’ section for further details) and imaged for 12 h at 10 min intervals; Scale bar 30 μm.

  4. 4.

    Time-lapse movie of karyokinesis with no cytokinesis (bi-nucleation) in mono-nucleated caErbb2 mice CMs.

    This is a representative time-lapse video of P7 caErbb2 mono-nucleated CMs performing karyokinesis but not cytokinesis (bi-nucleation). Heart cells were isolated from P7 neonatal caErbb2m mice, cultured in vitro for 48 h, then labelled with TMRE to identify CMs (see ‘Methods’ section for further details) and imaged for 12 h at 10 min intervals; Scale bar 30 μm.

  5. 5.

    Time-lapse movie of cytokinesis in bi-nucleated caErbb2 mice CMs.

    This is a representative time-lapse video of P7 caErbb2 bi-nucleated CMs performing cytokinesis. Heart cells were isolated from P7 neonatal caErbb2mice, cultured in vitro for 48 h, then labelled with TMRE to identify CMs (see ‘Methods’ section for further details) and imaged for 12 h at 10 min intervals; Scale bar 30 μm.

  6. 6.

    Time-lapse movie of CMs isolated from P7 ctrl mice.

    This is a representative time-lapse video of P7 ctrl CMs, showing no events of karyokinesis and cytokinesis. Heart cells were isolated from P7 ctrl mice, cultured in vitro for 48 h, then labelled with TMRE to identify CMs (see ‘Methods’ section for further details) and imaged for 12 h at 10 min intervals; Scale bar 30 μm.

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https://doi.org/10.1038/ncb3149

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