Osteologie 2021; 30(04): 339
DOI: 10.1055/s-0041-1736723
Abstract

Inhibition of Foxo3 during myogenic differentiation

Authors

  • E Rodenwaldt

    1   Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie der Universitätsmedizin Göttingen, Göttingen

  • B Gellhaus

    1   Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie der Universitätsmedizin Göttingen, Göttingen

  • M Gsaenger

    1   Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie der Universitätsmedizin Göttingen, Göttingen

  • R Kosinsky

    2   Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN, USA

  • O K Böker

    1   Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie der Universitätsmedizin Göttingen, Göttingen

  • F A Schilling

    1   Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie der Universitätsmedizin Göttingen, Göttingen

  • D Saul

    1   Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie der Universitätsmedizin Göttingen, Göttingen
    3   Mayo Clinic, Division of Endocrinology, Diabetes and Nutrition, Rochester, MN, USA
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Introduction

Patients suffering from disease-related (secondary) sarcopenia have been associated with an enhanced level of the transcription factor Foxo3 in skeletal muscle [1] [2] [3].

Sarcopenia is a progressive and generalized muscle disorder characterized by a decline in muscle mass and strength [4]. If left untreated, it can lead to increased falls, fractures [5] [6], mortality [7], reduced quality of life [8], as well as increased hospitalization rates and cost of care [9].

Foxo3 is one of several transcription factors of the highly conserved Forkhead-Box-Protein family [10]. As a downstream target of the PI3K/AKT pathway [11], Foxo3 plays an important role in protein turnover and muscle wasting [12]. Foxo3 therefore could pose to be a potential target of treatment for secondary sarcopenia.

The focus of this study is to reduce the Foxo3-expression in murine myoblasts in vitro and to analyze changes in myogenic differentiation.


 

Material and Methods

Foxo3 knockdown was achieved via single and double lipofection of Foxo3-siRNA. Subsequently, we differentiated myoblasts to myotubes. Length and width of myotubes were measured via light microscopy. Foxo3 mRNA analysis was performed by RT-qPCR.


 

Results

A Foxo3-mRNA knockdown has been confirmed 1-8 days after siRNA transfection with additional decrease of Atrogin1-mRNA. Morphologically, treated myotubes show an increase in length and width. Live/dead cell viability assay showed no significant difference between treated and untreated cells.


 

Conclusion

We were able to reach a significant Foxo3-mRNA knockdown of up to 82% after lipofection. As downstream member of the Foxo3 pathway, a 32% decrease of Atrogin1-mRNA was measured after lipofection. Morphologically, Foxo3-inhibited myotubes showed an increase in length and width.


 

 

Conflict of Interest

The authors declare no conflict of interest.

  • References

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  • 2 Mercken et al. Aging Cell 2013 (12), 645–651
  • 3 Parolo et al. PLoS ONE 2018 (13), e0194225
  • 4 Cruz-Jentoft et al. Age and Ageing 2019 (48), 16–31
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  • 7 De Buyser et al. Age Ageing 2016 (45), 603–608
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  • 9 Cawthon et al. The Journals of Gerontology: Series A 2017 (72), 1383–1389
  • 10 Hannenhalli et al. Nat Rev Genet 2009 (10), 233–240
  • 11 Brunet et al. Cell 1999 (96), 857–868
  • 12 Sacheck et al. American Journal of Physiology-Endocrinology and Metabolism 2004 287, E591–E601

Publication History

Article published online:
04 November 2021

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  • References

  • 1 Kneppers et al. Journal of the American Medical Directors Association 2017 (18) 637.e1–637.e11
  • 2 Mercken et al. Aging Cell 2013 (12), 645–651
  • 3 Parolo et al. PLoS ONE 2018 (13), e0194225
  • 4 Cruz-Jentoft et al. Age and Ageing 2019 (48), 16–31
  • 5 Bischoff-Ferrari et al. Osteoporos Int 2015 (10), 10.1007/s00198-015-3194-y
  • 6 Schaap et al. The Journals of Gerontology: Series A 2018 (73), 1199–1204
  • 7 De Buyser et al. Age Ageing 2016 (45), 603–608
  • 8 Rosenberg. J. Nutr. 1997 (127), 990S–991S
  • 9 Cawthon et al. The Journals of Gerontology: Series A 2017 (72), 1383–1389
  • 10 Hannenhalli et al. Nat Rev Genet 2009 (10), 233–240
  • 11 Brunet et al. Cell 1999 (96), 857–868
  • 12 Sacheck et al. American Journal of Physiology-Endocrinology and Metabolism 2004 287, E591–E601