Single-cell analysis uncovers that metabolic reprogramming by ErbB2 signaling is essential for cardiomyocyte proliferation in the regenerating heart

  1. Hessel Honkoop
  2. Dennis EM de Bakker
  3. Alla Aharonov
  4. Fabian Kruse
  5. Avraham Shakked
  6. Phong D Nguyen
  7. Cecilia de Heus
  8. Laurence Garric
  9. Mauro J Muraro
  10. Adam Shoffner
  11. Federico Tessadori
  12. Joshua Craiger Peterson
  13. Wendy Noort
  14. Alberto Bertozzi
  15. Gilbert Weidinger
  16. George Posthuma
  17. Dominic Grun
  18. Willem J van der Laarse
  19. Judith Klumperman
  20. Richard T Jaspers
  21. Kenneth D Poss
  22. Alexander van Oudenaarden
  23. Eldad Tzahor
  24. Jeroen Bakkers  Is a corresponding author
  1. Hubrecht Institute, Netherlands
  2. Weizmann Institute of Science, Israel
  3. University Medical Center Utrecht, Netherlands
  4. Duke University Medical Center, United States
  5. Vrije Universiteit Amsterdam, Netherlands
  6. Ulm University, Germany
  7. Max Planck Institute for Immunbiology and Epigenetics, Germany
  8. VU University Medical Center, Netherlands

Abstract

While the heart regenerates poorly in mammals, efficient heart regeneration occurs in zebrafish. Studies in zebrafish have resulted in a model in which preexisting cardiomyocytes dedifferentiate and reinitiate proliferation to replace the lost myocardium. To identify which processes occur in proliferating cardiomyocytes we have used a single-cell RNA-sequencing approach. We uncovered that proliferating border zone cardiomyocytes have very distinct transcriptomes compared to the nonproliferating remote cardiomyocytes and that they resemble embryonic cardiomyocytes. Moreover, these cells have reduced expression of mitochondrial genes and reduced mitochondrial activity, while glycolysis gene expression and glucose uptake are increased, indicative for metabolic reprogramming. Furthermore, we find that the metabolic reprogramming of border zone cardiomyocytes is induced by Nrg1/ErbB2 signaling and is important for their proliferation. This mechanism is conserved in murine hearts in which cardiomyocyte proliferation is induced by activating ErbB2 signaling. Together these results demonstrate that glycolysis regulates cardiomyocyte proliferation during heart regeneration.

Data availability

Sequencing data have been deposited in GEO under accession code GSE139218Other data generated during this study are included in the manuscript and supporting files

The following data sets were generated

Article and author information

Author details

  1. Hessel Honkoop

    Cardiac Development and Genetics, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  2. Dennis EM de Bakker

    Cardiac Development and Genetics, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  3. Alla Aharonov

    Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel
    Competing interests
    The authors declare that no competing interests exist.
  4. Fabian Kruse

    Cardiac Development and Genetics, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  5. Avraham Shakked

    Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel
    Competing interests
    The authors declare that no competing interests exist.
  6. Phong D Nguyen

    Cardiac Development and Genetics, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  7. Cecilia de Heus

    Department of Cell Biology, University Medical Center Utrecht, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  8. Laurence Garric

    Cardiac Development and Genetics, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  9. Mauro J Muraro

    Hubrecht Institute, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  10. Adam Shoffner

    Department of Cell Biology, Duke University Medical Center, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Federico Tessadori

    Cardiac Development and Genetics, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  12. Joshua Craiger Peterson

    Department of Human Movement Sciences, Vrije Universiteit Amsterdam, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  13. Wendy Noort

    Department of Human Movement Sciences, Vrije Universiteit Amsterdam, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  14. Alberto Bertozzi

    Institute of Biochemistry and Molecular Biology, Ulm University, Ulm, Germany
    Competing interests
    The authors declare that no competing interests exist.
  15. Gilbert Weidinger

    Institute of Biochemistry and Molecular Biology, Ulm University, Ulm, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3599-6760
  16. George Posthuma

    Department of Cell Biology, University Medical Center Utrecht, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  17. Dominic Grun

    Max Planck Institute for Immunbiology and Epigenetics, Freiburg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  18. Willem J van der Laarse

    Department of Physiology, Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  19. Judith Klumperman

    Department of Cell Biology, University Medical Center Utrecht, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  20. Richard T Jaspers

    Department of Human Movement Sciences, Vrije Universiteit Amsterdam, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6951-0952
  21. Kenneth D Poss

    Regeneration Next, Department of Cell Biology, Duke University Medical Center, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.
  22. Alexander van Oudenaarden

    Hubrecht Institute, Hubrecht Institute, Utrecht, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  23. Eldad Tzahor

    Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5212-9426
  24. Jeroen Bakkers

    Cardiac Development and Genetics, Hubrecht Institute, Utrecht, Netherlands
    For correspondence
    j.bakkers@hubrecht.eu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9418-0422

Funding

ERA-CVD (JCT2016-40-080)

  • Gilbert Weidinger
  • Eldad Tzahor
  • Jeroen Bakkers

NIH Clinical Center (R01 HL131319)

  • Kenneth D Poss

NIH Clinical Center (R01 HL136182)

  • Kenneth D Poss

Deutsche Forschungsgemeinschaft (251293561)

  • Gilbert Weidinger

Netherlands Heart Foundation NHS/CVON (Cobra3)

  • Jeroen Bakkers

European Molecular Biology Organization (ALTF1129-2015)

  • Phong D Nguyen

Human Frontier Science Program (LT001404/2017-L)

  • Phong D Nguyen

Dutch Research Council (016.186.017-3)

  • Phong D Nguyen

Deutsche Forschungsgemeinschaft (316249678)

  • Gilbert Weidinger

Deutsche Forschungsgemeinschaft (414077062)

  • Gilbert Weidinger

NIH Clinical Center (RO1 HL081674)

  • Kenneth D Poss

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All experiments were conducted in accordance with the ethical guidelines. Animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Royal Dutch Academy of Sciences (AVD801002016404), the state of Baden-Württemberg and the animal protection representative of Ulm University (Tierversuch 1352), Duke University (A057-18-02) and the Weizmann Institute (13240419-3).

Reviewing Editor

  1. Marianne E Bronner, California Institute of Technology, United States

Publication history

  1. Received: July 12, 2019
  2. Accepted: December 4, 2019
  3. Accepted Manuscript published: December 23, 2019 (version 1)
  4. Version of Record published: February 4, 2020 (version 2)

Copyright

© 2019, Honkoop et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Hessel Honkoop
  2. Dennis EM de Bakker
  3. Alla Aharonov
  4. Fabian Kruse
  5. Avraham Shakked
  6. Phong D Nguyen
  7. Cecilia de Heus
  8. Laurence Garric
  9. Mauro J Muraro
  10. Adam Shoffner
  11. Federico Tessadori
  12. Joshua Craiger Peterson
  13. Wendy Noort
  14. Alberto Bertozzi
  15. Gilbert Weidinger
  16. George Posthuma
  17. Dominic Grun
  18. Willem J van der Laarse
  19. Judith Klumperman
  20. Richard T Jaspers
  21. Kenneth D Poss
  22. Alexander van Oudenaarden
  23. Eldad Tzahor
  24. Jeroen Bakkers
(2019)
Single-cell analysis uncovers that metabolic reprogramming by ErbB2 signaling is essential for cardiomyocyte proliferation in the regenerating heart
eLife 8:e50163.
https://doi.org/10.7554/eLife.50163
  1. Further reading

Further reading

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    Experiments in zebrafish have shed new light on the relationship between development and regeneration in the heart.

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    The regeneration potential of the mammalian heart is incredibly limited, as cardiomyocyte proliferation ceases shortly after birth. β-adrenergic receptor (β-AR) blockade has been shown to improve heart functions in response to injury; however, the underlying mechanisms remain poorly understood. Here we inhibited β-AR signaling in the heart using metoprolol, a cardio-selective β blocker for β1-adrenergic receptor (β1-AR) to examine its role in heart maturation and regeneration in postnatal mice. We found that metoprolol enhanced cardiomyocyte proliferation and promoted cardiac regeneration post myocardial infarction, resulting in reduced scar formation and improved cardiac function. Moreover, the increased cardiomyocyte proliferation was also induced by the genetic deletion of Gnas, the gene encoding G protein alpha subunit (Gαs), a downstream effector of β-AR. Genome wide transcriptome analysis revealed that the Hippo-effector YAP, which is associated with immature cardiomyocyte proliferation, was upregulated in the cardiomyocytes of b-blocker treated and Gnas cKO hearts. Moreover, the increased YAP activity is modulated by RhoA signaling. Our pharmacological and genetic studies reveal that b1-AR-Gas-YAP signaling axis is involved in regulating postnatal cardiomyocyte proliferation. These results suggest that inhibiting b-AR-Gas signaling promotes the regenerative capacity and extends the cardiac regenerative window in juvenile mice by activating YAP-mediated transcriptional programs.