Dexamethasone Treatment at the Myoblast Stage Enhanced C2C12 Myocyte Differentiation

Han, Der‐Sheng; Yang, Wei-Shiung; Kao, Tung‐Wei

doi:10.7150/ijms.18427

“…www.nature.com/scientificreports www.nature.com/scientificreports/ differentiation 58 . Finally, dexamethasone administration to myotubes produces an atrophic effect with increased expression of atrogin-1 and a decreased protein content of MyHC [58][59][60] . Reconciling the conclusions of different reports is made difficult by the heterogeneity of GCs activity especially when tested on different cell types, primary or stable cell lines.…”

Section: Scientific Reports |mentioning

confidence: 99%

Janus effect of glucocorticoids on differentiation of muscle fibro/adipogenic progenitors

Perpetuini

¹

,

Giuliani

²

,

Reggio

³

et al. 2020

Sci Rep

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Muscle resident fibro-adipogenic progenitors (FAPs), support muscle regeneration by releasing cytokines that stimulate the differentiation of myogenic stem cells. However, in non-physiological contexts (myopathies, atrophy, aging) FAPs cause fibrotic and fat infiltrations that impair muscle function. We set out to perform a fluorescence microscopy-based screening to identify compounds that perturb the differentiation trajectories of these multipotent stem cells. From a primary screen of 1,120 FDA/EMA approved drugs, we identified 34 compounds as potential inhibitors of adipogenic differentiation of FAPs isolated from the murine model (mdx) of Duchenne muscular dystrophy (DMD). The hit list from this screen was surprisingly enriched with compounds from the glucocorticoid (GCs) chemical class, drugs that are known to promote adipogenesis in vitro and in vivo. To shed light on these data, three GCs identified in our screening efforts were characterized by different approaches. We found that like dexamethasone, budesonide inhibits adipogenesis induced by insulin in sub-confluent FAPs. However, both drugs have a pro-adipogenic impact when the adipogenic mix contains factors that increase the concentration of cAMP. Gene expression analysis demonstrated that treatment with glucocorticoids induces the transcription of Gilz/Tsc22d3, an inhibitor of the adipogenic master regulator PPARγ, only in anti-adipogenic conditions. Additionally, alongside their anti-adipogenic effect, GCs are shown to promote terminal differentiation of satellite cells. Both the anti-adipogenic and pro-myogenic effects are mediated by the glucocorticoid receptor and are not observed in the presence of receptor inhibitors. Steroid administration currently represents the standard treatment for DMD patients, the rationale being based on their anti-inflammatory effects. The findings presented here offer new insights on additional glucocorticoid effects on muscle stem cells that may affect muscle homeostasis and physiology.In muscular dystrophies the degeneration of muscle tissue is initially compensated by efficient regeneration that neutralize muscle loss 1 . However, over time, the regenerative potential in muscles of patients affected by myopathies is impaired and myofibers repair is curbed by the formation of fibrotic scars and fat infiltrations, ultimately leading to decreased muscle function 2 . Muscle fibro-adipogenic progenitors play an important role in these processes. FAPs are muscle mesenchymal stem cells residing in the muscle fibers interstitial space. FAPs express the SCA1, CD34 and PDGFRα (CD140a) antigens while they are negative for the hematopoietic and endothelial markers, CD45 and CD31, and for the satellite marker α7 integrin (ITGA7) 2-4 . FAPs contribute to muscle regeneration by secreting IGF-1 and IL-6 3 , by facilitating the clearance of necrotic debris 5 and by promoting the formation of extra-cellular matrix 6 . In addition to this pro-regenerative roles, FAPs are responsible for the formation of ectopic tissue infiltrations ...

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“…In addition and relevant for their impact on treatment of muscle disorders glucocorticoids inhibit myogenesis by inducing the expression of the glucocorticoid-induced leucine zipper (Gilz) which in turn inhibits MyoD function 62 . On the other hand, somewhat in contrast, the exposure of C2C12 myoblasts to dexamethasone causes an increase in proliferation rate and terminal myogenic differentiation 63 . Finally, dexamethasone administration to myotubes produces an atrophic effect with increased expression of atrogin-1 and a decreased protein content of MyHC [64][65][66] .…”

Section: Discussionmentioning

confidence: 92%

“…On the other hand, somewhat in contrast, the exposure of C2C12 myoblasts to dexamethasone causes an increase in proliferation rate and terminal myogenic differentiation 63 . Finally, dexamethasone administration to myotubes produces an atrophic effect with increased expression of atrogin-1 and a decreased protein content of MyHC [64][65][66] .…”

Section: Discussionmentioning

confidence: 92%

“…In addition, several GCs such as dexamethasone and prednisolone, can exert positive or negative effects on myogenic cell lines, depending on the stage of administration. The exposure of C2C12 myoblast to dexamethasone causes an increase in terminal differentiation 61 . Similarly, we observed that budesonide treatment of primary mdx satellite cells also promotes terminal differentiation.…”

Section: Discussionmentioning

confidence: 99%

“…It was proposed that steroids could exert their beneficial effects on DMD patients by inhibiting muscle proteolysis 67 . However, dexamethasone administration to myotubes produces an atrophic effect with increased expression of atrogin-1 [62][63][64] and muscle atrophy is one of the main side effects of prolonged GCs treatment 68 .…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Janus effect of glucocorticoids on differentiation of muscle fibro/adipogenic progenitors

Perpetuini

¹

,

Reggio

²

,

Cerretani

³

et al. 2019

Preprint

View full text Add to dashboard Cite

Muscle resident fibro-adipogenic progenitors (FAPs), support muscle regeneration by releasing cytokines that stimulate the differentiation of myogenic stem cells. However, in non-physiological contexts (myopathies, atrophy and aging) FAPs cause fibrotic and fat infiltrations that impair muscle function. We set out to perform a fluorescence microscopy-based screening to identify compounds that perturb the differentiation trajectories of these multipotent stem cells. From a primary screening of 1120 FDA/EMA approved drugs, we identified 34 compounds as potential inhibitors of adipogenic differentiation of FAPs isolated from the murine model (mdx) of Duchenne muscular dystrophy (DMD). To our surprise we found the hit list enriched for the glucocorticoid chemical class (GCs). Glucorticoids are often described as drugs that promote adipogenesis in vitro and in vivo. To shed light on conflicting reports we further characterized by different approaches three GCs identified in our screening focusing on budesonide a potent inhibitor of adipogenesis. We found that, similarly to dexamethasone, budesonide inhibits adipogenesis induced by insulin on subconfuent FAPs. However, both drugs have a proadipogenic impact when the adipogenic mix contains factors that increase the concentration of cAMP. By gene expression analysis we observed that treatment with glucocorticoids induces the transcription of Gilz/Tsc22d3, an inhibitor of the adipogenic master regulator PPARγ, only in antiadipogenic conditions. Finally, we noticed that alongside their anti-adipogenic effect, GCs promote terminal differentiation of satellite cells. Both the pro-adipogenic and pro-myogenic effects are mediated by the glucocorticoid receptor as they are not observed in the presence of receptor inhibitor. Steroid administration currently represents the standard treatment of DMD patients, the rationale being based on their antiinflammatory effects. The findings presented here offer new insights on additional glucocorticoid effects on muscle stem cells that may affect muscle homeostasis and physiology.

show abstract

Skeletal Muscle Constructs Engineered from Human Embryonic Stem Cell Derived Myogenic Progenitors Exhibit Enhanced Contractile Forces When Differentiated in a Medium Containing EGM‐2 Supplements

Xu

¹

,

Zhang

²

,

Perlingeiro

³

2019

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Three-dimensional skeletal muscle constructs engineered from myogenic progenitors derived from human pluripotent stem cells (hPSCs) have a wide range of applications, but to date such constructs generate lower specific tetanic force than adult human muscles. Methods enhancing functional muscle differentiation and force generation of these constructs are highly desirable. The finding of this study is that addition of the supplements in the EGM-2 medium to the myogenic differentiation medium could substantially enhance contractile force generation. For constructs differentiated for four weeks, addition of the EGM-2 supplements in the first two weeks leads to 10-fold and 7-fold increases in twitch and tetanic forces, respectively. The specific tetanic force generated by these constructs is 33 mN mm-2 , which is significantly higher than previously reported. These constructs show wider myotubes and higher gene expression levels for all skeletal muscle-specific myosin heavy chain isoforms, suggesting that a more mature differentiation stage of the cells underlies the greater contractile force generation. The constructs exposed to these supplements for four weeks do not generate as high contractile forces, suggesting that prolonged treatment is not beneficial. These results suggest that temporal conditioning with the EGM-2 supplements assists functional development of hPSC-derived skeletal muscle constructs.

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Dexamethasone Treatment at the Myoblast Stage Enhanced C2C12 Myocyte Differentiation

Cited by 32 publications

References 27 publications

Janus effect of glucocorticoids on differentiation of muscle fibro/adipogenic progenitors

Janus effect of glucocorticoids on differentiation of muscle fibro/adipogenic progenitors

Janus effect of glucocorticoids on differentiation of muscle fibro/adipogenic progenitors

Skeletal Muscle Constructs Engineered from Human Embryonic Stem Cell Derived Myogenic Progenitors Exhibit Enhanced Contractile Forces When Differentiated in a Medium Containing EGM‐2 Supplements

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