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TNF-α and IGF1 modify the microRNA signature in skeletal muscle cell differentiation.

Meyer SU, Thirion C, Polesskaya A, Bauersachs S, Kaiser S, Krause S, Pfaffl MW - Cell Commun. Signal (2015)

Bottom Line: Results reveal that i) TNF-α and IGF1 regulate miRNA expression during skeletal muscle cell differentiation in vitro, ii) microRNA targets can mediate the negative effect of TNF-α on fusion capacity of skeletal myoblasts by targeting genes associated with axon guidance, MAPK signalling, focal adhesion, and neurotrophin signalling pathway, iii) inhibition of miR-155 in combination with overexpression of miR-503 partially abrogates the inhibitory effect of TNF-α on myotube formation, and iv) MAPK/ERK inhibition might participate in modulating the effect of TNF-α and IGF1 on miRNA abundance.The inhibitory effects of TNF-α or the growth promoting effects of IGF1 on skeletal muscle differentiation include the deregulation of known muscle-regulatory miRNAs as well as miRNAs which have not yet been associated with skeletal muscle differentiation or response to TNF-α or IGF1.This study indicates that miRNAs are mediators of the inhibitory effect of TNF-α on myoblast differentiation.

View Article: PubMed Central - PubMed

Affiliation: Physiology Weihenstephan, ZIEL Research Center for Nutrition and Food Sciences, Technische Universität München, Weihenstephaner Berg 3, D-85354, Freising, Germany. meyers@wzw.tum.de.

ABSTRACT

Background: Elevated levels of the inflammatory cytokine TNF-α are common in chronic diseases or inherited or degenerative muscle disorders and can lead to muscle wasting. By contrast, IGF1 has a growth promoting effect on skeletal muscle. The molecular mechanisms mediating the effect of TNF-α and IGF1 on muscle cell differentiation are not completely understood. Muscle cell proliferation and differentiation are regulated by microRNAs (miRNAs) which play a dominant role in this process. This study aims at elucidating how TNF-α or IGF1 regulate microRNA expression to affect myoblast differentiation and myotube formation.

Results: In this study, we analyzed the impact of TNF-α or IGF1 treatment on miRNA expression in myogenic cells. Results reveal that i) TNF-α and IGF1 regulate miRNA expression during skeletal muscle cell differentiation in vitro, ii) microRNA targets can mediate the negative effect of TNF-α on fusion capacity of skeletal myoblasts by targeting genes associated with axon guidance, MAPK signalling, focal adhesion, and neurotrophin signalling pathway, iii) inhibition of miR-155 in combination with overexpression of miR-503 partially abrogates the inhibitory effect of TNF-α on myotube formation, and iv) MAPK/ERK inhibition might participate in modulating the effect of TNF-α and IGF1 on miRNA abundance.

Conclusions: The inhibitory effects of TNF-α or the growth promoting effects of IGF1 on skeletal muscle differentiation include the deregulation of known muscle-regulatory miRNAs as well as miRNAs which have not yet been associated with skeletal muscle differentiation or response to TNF-α or IGF1. This study indicates that miRNAs are mediators of the inhibitory effect of TNF-α on myoblast differentiation. We show that intervention at the miRNA level can ameliorate the negative effect of TNF-α by promoting myoblast differentiation. Moreover, we cautiously suggest that TNF-α or IGF1 modulate the miRNA biogenesis of some miRNAs via MAPK/ERK signalling. Finally, this study identifies indicative biomarkers of myoblast differentiation and cytokine influence and points to novel RNA targets.

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Principal component analyses of miRNA in early skeletal myoblast differentiation and TNF-α or IGF1 treatment. Principal component analyses of murine miRNA expression profiling data after 24 h of induction differentiation and TNF-α or IGF1 treatment. Principal component analysis reveals separation of treatment groups for (A) microarray and (B) qPCR data. Dynamic principal component analysis (group selection myoblasts) identifies the most relevant subset of miRNAs which can describe the treatment effects and separate the effects by principal components for (C) microarray and (D) qPCR data. Axes depict principal component 1 (PC 1), principal component 2 (PC 2), and principal component 3 (PC 3).
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Fig2: Principal component analyses of miRNA in early skeletal myoblast differentiation and TNF-α or IGF1 treatment. Principal component analyses of murine miRNA expression profiling data after 24 h of induction differentiation and TNF-α or IGF1 treatment. Principal component analysis reveals separation of treatment groups for (A) microarray and (B) qPCR data. Dynamic principal component analysis (group selection myoblasts) identifies the most relevant subset of miRNAs which can describe the treatment effects and separate the effects by principal components for (C) microarray and (D) qPCR data. Axes depict principal component 1 (PC 1), principal component 2 (PC 2), and principal component 3 (PC 3).

Mentions: The regulation of miRNA expression by TNF-α and IGF1 treatment of differentiating murine skeletal muscle cells appears in separation of treatment groups by hierarchical cluster analysis (Additional file 6A and B) or principal component analysis (Figure 2A,B). The distance between treatment groups depended on the profiling platform used as well as the clustering approach. We applied dynamic principal component analysis to identify the most relevant miRNAs explaining our observations (Figure 2C,D). However, separation of treatment groups became less clear or even disappeared between control myotubes and TNF-α or IGF1-treated myotubes, whereas myoblasts always separated from the differentiation effects. Five miRNAs which are in the subset of miRNAs derived from dynamic principal component analysis were identified by the miRNA microarray as well as the qPCR-platform: mmu-miR-133b-3p, mmu-miR-188-3p, mmu-miR-206-3p, mmu-miR-335-3p, mmu-miR-351-3p (Table 2). In conclusion, variable selection by dynamic principal component analysis revealed distinct miRNAs identified on different platforms which might be indicative biomarkers for myoblast differentiation.Figure 2


TNF-α and IGF1 modify the microRNA signature in skeletal muscle cell differentiation.

Meyer SU, Thirion C, Polesskaya A, Bauersachs S, Kaiser S, Krause S, Pfaffl MW - Cell Commun. Signal (2015)

Principal component analyses of miRNA in early skeletal myoblast differentiation and TNF-α or IGF1 treatment. Principal component analyses of murine miRNA expression profiling data after 24 h of induction differentiation and TNF-α or IGF1 treatment. Principal component analysis reveals separation of treatment groups for (A) microarray and (B) qPCR data. Dynamic principal component analysis (group selection myoblasts) identifies the most relevant subset of miRNAs which can describe the treatment effects and separate the effects by principal components for (C) microarray and (D) qPCR data. Axes depict principal component 1 (PC 1), principal component 2 (PC 2), and principal component 3 (PC 3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4325962&req=5

Fig2: Principal component analyses of miRNA in early skeletal myoblast differentiation and TNF-α or IGF1 treatment. Principal component analyses of murine miRNA expression profiling data after 24 h of induction differentiation and TNF-α or IGF1 treatment. Principal component analysis reveals separation of treatment groups for (A) microarray and (B) qPCR data. Dynamic principal component analysis (group selection myoblasts) identifies the most relevant subset of miRNAs which can describe the treatment effects and separate the effects by principal components for (C) microarray and (D) qPCR data. Axes depict principal component 1 (PC 1), principal component 2 (PC 2), and principal component 3 (PC 3).
Mentions: The regulation of miRNA expression by TNF-α and IGF1 treatment of differentiating murine skeletal muscle cells appears in separation of treatment groups by hierarchical cluster analysis (Additional file 6A and B) or principal component analysis (Figure 2A,B). The distance between treatment groups depended on the profiling platform used as well as the clustering approach. We applied dynamic principal component analysis to identify the most relevant miRNAs explaining our observations (Figure 2C,D). However, separation of treatment groups became less clear or even disappeared between control myotubes and TNF-α or IGF1-treated myotubes, whereas myoblasts always separated from the differentiation effects. Five miRNAs which are in the subset of miRNAs derived from dynamic principal component analysis were identified by the miRNA microarray as well as the qPCR-platform: mmu-miR-133b-3p, mmu-miR-188-3p, mmu-miR-206-3p, mmu-miR-335-3p, mmu-miR-351-3p (Table 2). In conclusion, variable selection by dynamic principal component analysis revealed distinct miRNAs identified on different platforms which might be indicative biomarkers for myoblast differentiation.Figure 2

Bottom Line: Results reveal that i) TNF-α and IGF1 regulate miRNA expression during skeletal muscle cell differentiation in vitro, ii) microRNA targets can mediate the negative effect of TNF-α on fusion capacity of skeletal myoblasts by targeting genes associated with axon guidance, MAPK signalling, focal adhesion, and neurotrophin signalling pathway, iii) inhibition of miR-155 in combination with overexpression of miR-503 partially abrogates the inhibitory effect of TNF-α on myotube formation, and iv) MAPK/ERK inhibition might participate in modulating the effect of TNF-α and IGF1 on miRNA abundance.The inhibitory effects of TNF-α or the growth promoting effects of IGF1 on skeletal muscle differentiation include the deregulation of known muscle-regulatory miRNAs as well as miRNAs which have not yet been associated with skeletal muscle differentiation or response to TNF-α or IGF1.This study indicates that miRNAs are mediators of the inhibitory effect of TNF-α on myoblast differentiation.

View Article: PubMed Central - PubMed

Affiliation: Physiology Weihenstephan, ZIEL Research Center for Nutrition and Food Sciences, Technische Universität München, Weihenstephaner Berg 3, D-85354, Freising, Germany. meyers@wzw.tum.de.

ABSTRACT

Background: Elevated levels of the inflammatory cytokine TNF-α are common in chronic diseases or inherited or degenerative muscle disorders and can lead to muscle wasting. By contrast, IGF1 has a growth promoting effect on skeletal muscle. The molecular mechanisms mediating the effect of TNF-α and IGF1 on muscle cell differentiation are not completely understood. Muscle cell proliferation and differentiation are regulated by microRNAs (miRNAs) which play a dominant role in this process. This study aims at elucidating how TNF-α or IGF1 regulate microRNA expression to affect myoblast differentiation and myotube formation.

Results: In this study, we analyzed the impact of TNF-α or IGF1 treatment on miRNA expression in myogenic cells. Results reveal that i) TNF-α and IGF1 regulate miRNA expression during skeletal muscle cell differentiation in vitro, ii) microRNA targets can mediate the negative effect of TNF-α on fusion capacity of skeletal myoblasts by targeting genes associated with axon guidance, MAPK signalling, focal adhesion, and neurotrophin signalling pathway, iii) inhibition of miR-155 in combination with overexpression of miR-503 partially abrogates the inhibitory effect of TNF-α on myotube formation, and iv) MAPK/ERK inhibition might participate in modulating the effect of TNF-α and IGF1 on miRNA abundance.

Conclusions: The inhibitory effects of TNF-α or the growth promoting effects of IGF1 on skeletal muscle differentiation include the deregulation of known muscle-regulatory miRNAs as well as miRNAs which have not yet been associated with skeletal muscle differentiation or response to TNF-α or IGF1. This study indicates that miRNAs are mediators of the inhibitory effect of TNF-α on myoblast differentiation. We show that intervention at the miRNA level can ameliorate the negative effect of TNF-α by promoting myoblast differentiation. Moreover, we cautiously suggest that TNF-α or IGF1 modulate the miRNA biogenesis of some miRNAs via MAPK/ERK signalling. Finally, this study identifies indicative biomarkers of myoblast differentiation and cytokine influence and points to novel RNA targets.

Show MeSH
Related in: MedlinePlus