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Ghrelin knockout mice display defective skeletal muscle regeneration and impaired satellite cell self-renewal

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Abstract

Purpose

Muscle regeneration depends on satellite cells (SCs), quiescent precursors that, in consequence of injury or pathological states such as muscular dystrophies, activate, proliferate, and differentiate to repair the damaged tissue. A subset of SCs undergoes self-renewal, thus preserving the SC pool and its regenerative potential. The peptides produced by the ghrelin gene, i.e., acylated ghrelin (AG), unacylated ghrelin (UnAG), and obestatin (Ob), affect skeletal muscle biology in several ways, not always with overlap** effects. In particular, UnAG and Ob promote SC self-renewal and myoblast differentiation, thus fostering muscle regeneration.

Methods

To delineate the endogenous contribution of preproghrelin in muscle regeneration, we evaluated the repair process in Ghrl−/− mice upon CTX-induced injury.

Results

Although muscles from Ghrl−/− mice do not visibly differ from WT muscles in term of weight, structure, and SCs content, muscle regeneration after CTX-induced injury is impaired in Ghrl−/− mice, indicating that ghrelin-derived peptides actively participate in muscle repair. Remarkably, the lack of ghrelin gene impacts SC self-renewal during regeneration.

Conclusions

Although we cannot discern the specific Ghrl-derived peptide responsible for such activities, these data indicate that Ghrl contributes to a proper muscle regeneration.

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References

  1. P. Seale, L.A. Sabourin, A. Girgis-Gabardo, A. Mansouri, P. Gruss, M.A. Rudnicki, Pax7 is required for the specification of myogenic satellite cells. Cell 102, 777–786 (2000)

    Article  CAS  PubMed  Google Scholar 

  2. U. Gurriarán-Rodríguez, I. Santos-zas, O. Al-massadi, C.S. Mosteiro, R. Nogueiras, A.B. Crujeiras, L.M. Seoane, J. Señarís, R. Gallego, F. Felipe, Y. Pazos, J.P. Camiña, The obestatin/GPR39 system is up-regulated by muscle injury and functions as an autocrine regenerative system. J. Biol. Chem. 287, 38379–38389 (2012)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. S. Reano, E. Angelino, M. Ferrara, V. Malacarne, H. Sustova, O. Sabry, E. Agosti, S. Clerici, G. Ruozi, L. Zentilin, F. Prodam, S. Geuna, M. Giacca, A. Graziani, N. Filigheddu, Unacylated ghrelin enhances satellite cell function and relieves the dystrophic phenotype in Duchenne muscular dystrophy mdx model. Stem Cells 35, 1733–1746 (2017)

    Article  CAS  PubMed  Google Scholar 

  4. U. Gurriarán-Rodríguez, I. Santos-zas, J. González-sánchez, D. Beiroa, V. Moresi, C.S. Mosteiro, W. Lin, J.E. Viñuela, J. Señarís, T. García-caballero, F.F. Casanueva, R. Nogueiras, R. Gallego, J. Renaud, S. Adamo, Y. Pazos, J.P. Camiña, Action of obestatin in skeletal muscle repair: stem cell expansion, muscle growth, and microenvironment remodeling. Mol. Ther. 23, 1003–1021 (2015)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. G. Ruozi, F. Bortolotti, A. Falcione, M.D. Ferro, L. Ukovich, A. Macedo, L. Zentilin, N. Filigheddu, G.G. Cappellari, G. Baldini, M. Zweyer, R. Barazzoni, A. Graziani, S. Zacchigna, M. Giacca, AAV-mediated in vivo functional selection of tissue-protective factors against ischaemia. Nat. Commun. 6(6), 7388 (2015)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. G.G. Togliatto, A. Trombetta, P. Dentelli, P. Cotogni, A. Rosso, H. Matthias, R. Granata, E. Ghigo, M.F. Brizzi, G. Togliatto, A. Trombetta, P. Dentelli, P. Cotogni, A. Rosso, Unacylated ghrelin promotes skeletal muscle regeneration following hindlimb ischemia via SOD-2-mediated miR-221/222 expression. J. Am. Heart Assoc. 2, e000376 (2013)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. N.Filigheddu, V.F.Gnocchi, M.Coscia, M.Cappelli, P.E.Porporato, R.Taulli, S. Traini, G.Baldanzi, F.Chianale, S.Cutrupi, E.Arnoletti, C.Ghe, A.Fubini, N.Surico, F.Sinigaglia, C.Ponzetto, G.Muccioli, T.Crepaldi, A.Graziani, Ghrelin and des-acyl ghrelin promote differentiation and fusion of C2C12 skeletal muscle cells. Mol. Biol. Cell 18, 986–994 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. I. Santos-Zas, U. Gurriarán-Rodríguez, T. Cid-Díaz, G. Figueroa, J. González-Sánchez, M. Bouzo-Lorenzo, C.S. Mosteiro, J. Señarís, F.F. Casanueva, X. Casabiell, R. Gallego, Y. Pazos, V. Mouly, J.P. Camiña, β-Arrestin scaffolds and signaling elements essential for the obestatin/GPR39 system that determine the myogenic program in human myoblast cells. Cell. Mol. Life. Sci. 73, 617–635 (2016)

    Article  CAS  PubMed  Google Scholar 

  9. A.P. Yu, X.M. Pei, T.K. Sin, S.P. Yip, B.Y. Yung, L.W. Chan, C.S. Wong, P.M. Siu, Acylated and unacylated ghrelin inhibit doxorubicin-induced apoptosis in skeletal muscle. Acta Physiol. 211, 201–213 (2014)

    Article  CAS  Google Scholar 

  10. A. Balasubramaniam, R. Joshi, C. Su, L.A. Friend, S. Sheriff, R.J. Kagan, J.H. James, Ghrelin inhibits skeletal muscle protein breakdown in rats with thermal injury through normalizing elevated expression of E3 ubiquitin ligases MuRF1 and MAFbx. Am. J. Physiol. Regul. Integr. Comp. Physiol. 296, R893 (2009)

    Article  CAS  PubMed  Google Scholar 

  11. S. Sheriff, N. Kadeer, R. Joshi, L. Ann, J.H. James, A. Balasubramaniam, Des-acyl ghrelin exhibits pro-anabolic and anti-catabolic effects on C2C12 myotubes exposed to cytokines and reduces burn-induced muscle proteolysis in rats. Mol. Cell. Endocrinol. 351, 286–295 (2012)

    Article  CAS  PubMed  Google Scholar 

  12. P.E. Porporato, N. Filigheddu, S. Reano, M. Ferrara, E. Angelino, V.F. Gnocchi, F. Prodam, G. Ronchi, S. Fagoonee, M. Fornaro, F. Chianale, G. Baldanzi, N. Surico, F. Sinigaglia, I. Perroteau, R.G. Smith, Y. Sun, S. Geuna, A. Graziani, Acylated and unacylated ghrelin impair skeletal muscle atrophy in mice. J. Clin. Invest. 123, 611–622 (2013)

    CAS  PubMed  PubMed Central  Google Scholar 

  13. M. Tamaki, A. Hagiwara, K. Miyashita, S. Wakino, H. Inoue, K. Fujii, C. Fujii, M. Sato, M. Mitsuishi, A. Muraki, K. Hayashi, T. Doi, H. Itoh, Improvement of physical decline through combined effects of muscle enhancement and mitochondrial activation by a gastric hormone ghrelin in male 5/6Nx CKD model mice. Endocrinology 156, 3638–3648 (2015)

    Article  CAS  PubMed  Google Scholar 

  14. G.G. Cappellari, A. Semolic, G. Ruozi, P. Vinci, G. Guarnieri, F. Bortolotti, D. Barbetta, M. Zanetti, M. Giacca, R. Barazzoni, Unacylated ghrelin normalizes skeletal muscle oxidative stress and prevents muscle catabolism by enhancing tissue mitophagy in experimental chronic kidney disease. FASEB J. 31, 5159–5171 (2017)

    Article  CAS  Google Scholar 

  15. X. Zeng, S. Chen, Y. Yang, Z. Ke, Acylated and unacylated ghrelin inhibit atrophy in myotubes co-cultured with colon carcinoma cells. Oncotarget 8, 72872–72885 (2017)

    PubMed  PubMed Central  Google Scholar 

  16. K. Koshinaka, K. Toshinai, A. Mohammad, K. Noma, M. Oshikawa, H. Ueno, H. Yamaguchi, M. Nakazato, Therapeutic potential of ghrelin treatment for unloading-induced muscle atrophy in mice. Biochem. Biophys. Res. Commun. 412, 296–301 (2011)

    Article  CAS  PubMed  Google Scholar 

  17. M. Sugiyama, A. Yamaki, M. Furuya, N. Inomata, Y. Minamitake, K. Ohsuye, K. Kangawa, Ghrelin improves body weight loss and skeletal muscle catabolism associated with angiotensin II-induced cachexia in mice. Regul. Pept. 178, 21–28 (2012)

    Article  CAS  PubMed  Google Scholar 

  18. H. Tsubouchi, S. Yanagi, A. Miura, N. Matsumoto, K. Kangawa, M. Nakazato, Ghrelin relieves cancer cachexia associated with the development of lung adenocarcinoma in mice. Eur. J. Pharmacol. 743, 1–10 (2014)

    Article  CAS  PubMed  Google Scholar 

  19. J.Chen, A.Splenser, B.Guillory, J.Luo, M.Mendiratta, B.Belinova, T.Halder, G.Zhang, Y.Li, J.M.Garcia, Ghrelin prevents tumour- and cisplatin-induced muscle wasting: characterization of multiple mechanisms involved. J Cachexia Sarcopenia Muscle 6, 132–143 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  20. G.G. Cappellari, M. Zanetti, A. Semolic, P. Vinci, G. Ruozi, A. Falcione, N. Filigheddu, G. Guarnieri, A. Graziani, M. Giacca, R. Barazzoni, Unacylated ghrelin reduces skeletal muscle reactive oxygen species generation and inflammation and prevents high-fat diet-induced hyperglycemia and whole-body insulin resistance in rodents. Diabetes 65, 874–886 (2016)

    Article  CAS  Google Scholar 

  21. R. Hassouna, P. Zizzari, C. Tomasetto, J.D. Veldhuis, O. Fiquet, A. Labarthe, J. Cognet, F. Steyn, C. Chen, J. Epelbaum, V. Tolle, An early reduction in GH peak amplitude in preproghrelin-deficient male mice has a minor impact on linear growth. Endocrinology 155, 3561–3571 (2014)

    Article  CAS  PubMed  Google Scholar 

  22. K.L. Shea, W. **ang, V.S. Laporta, J.D. Licht, C. Keller, M.A. Basson, A.S. Brack, Sprouty1 regulates reversible quiescence of a self-renewing adult muscle stem cell pool during regeneration. Cell Stem Cell 6, 117–129 (2010)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. F. Prodam, N. Filigheddu, Ghrelin gene products in acute and chronic inflammation. Arch. Immunol. Ther. Exp. (Warsz). 62, 369–384 (2014)

    Article  CAS  PubMed  Google Scholar 

  24. H. Yin, F. Price, M.A. Rudnicki, Satellite cells and the muscle stem cell niche. Physiol. Rev. 93, 23–67 (2013)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This study was supported by research grants from the Muscular Dystrophy Association (grant MDA294617 to NF and AG), Association Française contre les Myopathies (Grant 16437 to AG), AIRC (to AG), and Fondazione Cariplo (Grant 2015_0634 to NF).

Author information

Author notes

  1. These authors contributed equally: Elia Angelino, Simone Reano.

  2. These authors jointly supervised this work: Andrea Graziani, Nicoletta Filigheddu.

  3. Elia Angelino, Simone Reano, Michele Ferrara, Marilisa De Feudis, Hana Sustova, Emanuela Agosti, Andrea Graziani, and Nicoletta Filigheddu are members of Istituto Interuniversitario di Miologia (IIM).

Authors and Affiliations

  1. Department of Translational Medicine, University of Piemonte Orientale, Novara, Italy

    Elia Angelino, Simone Reano, Alessandro Bollo, Michele Ferrara, Marilisa De Feudis, Hana Sustova, Emanuela Agosti, Sara Clerici, Andrea Graziani & Nicoletta Filigheddu

  2. Università Vita-Salute San Raffaele, Milano, Italy

    Elia Angelino, Alessandro Bollo, Michele Ferrara, Sara Clerici & Andrea Graziani

  3. Department of Health Sciences, University of Piemonte Orientale, Novara, Italy

    Flavia Prodam

  4. IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire - Université de Strasbourg, Illkirch, France

    Catherine-Laure Tomasetto

Authors
  1. Elia Angelino
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  2. Simone Reano
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  12. Nicoletta Filigheddu
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Corresponding authors

Correspondence to Andrea Graziani or Nicoletta Filigheddu.

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Conflict of interest

A.G. is a consultant to Helsinn (Lugano, Switzerland), N.F. is a consultant to Lyric Pharmaceuticals (South San Francisco, CA, USA).

Ethical approval

This article does not contain any studies with human participants performed by any of the authors. Animal experiments were performed according to procedures approved by the Institutional Animal Care and Use Committee at the University of Piemonte Orientale.

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Angelino, E., Reano, S., Bollo, A. et al. Ghrelin knockout mice display defective skeletal muscle regeneration and impaired satellite cell self-renewal. Endocrine 62, 129–135 (2018). https://doi.org/10.1007/s12020-018-1606-4

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