Limits...
Extracellular microRNAs are dynamic non-vesicular biomarkers of muscle turnover.

Roberts TC, Godfrey C, McClorey G, Vader P, Briggs D, Gardiner C, Aoki Y, Sargent I, Morgan JE, Wood MJ - Nucleic Acids Res. (2013)

Bottom Line: Expression of the myogenic miRNA, miR-206 and the myogenic transcription factor myogenin in the tibialis anterior muscle were found to positively correlate with serum dystromiR levels, suggesting that extracellular miRNAs are indicators of the regenerative status of the musculature.Only a minority of serum dystromiRs were found in extracellular vesicles, whereas the majority were protected from serum nucleases by association with protein/lipoprotein complexes.In conclusion, extracellular miRNAs are dynamic indices of pathophysiological processes in skeletal muscle.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3QX, UK, Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037, USA, Dubowitz Neuromuscular Centre, UCL Institute of Child Health, 30 Guildford Street, London, WC1N 1EH, UK and Nuffield Department of Obstetrics and Gynaecology, University of Oxford, Level 3, Women's Centre, John Radcliffe Hospital, Oxford, OX3 9DU, UK.

ABSTRACT
Extracellular microRNAs (miRNAs) are promising biomarkers of the inherited muscle wasting condition Duchenne muscular dystrophy, as they allow non-invasive monitoring of either disease progression or response to therapy. In this study, serum miRNA profiling reveals a distinct extracellular miRNA signature in dystrophin-deficient mdx mice, which shows profound dose-responsive restoration following dystrophin rescue. Extracellular dystrophy-associated miRNAs (dystromiRs) show dynamic patterns of expression that mirror the progression of muscle pathology in mdx mice. Expression of the myogenic miRNA, miR-206 and the myogenic transcription factor myogenin in the tibialis anterior muscle were found to positively correlate with serum dystromiR levels, suggesting that extracellular miRNAs are indicators of the regenerative status of the musculature. Similarly, extracellular dystromiRs were elevated following experimentally-induced skeletal muscle injury and regeneration in non-dystrophic mice. Only a minority of serum dystromiRs were found in extracellular vesicles, whereas the majority were protected from serum nucleases by association with protein/lipoprotein complexes. In conclusion, extracellular miRNAs are dynamic indices of pathophysiological processes in skeletal muscle.

Show MeSH

Related in: MedlinePlus

Serum dystromiR abundance correlates with expression of regeneration factors in the TA. TA muscles from mdx and Pip6e-PMO-treated mdx mice were harvested at various time points, and the expression of the myogenic transcription factors (A) Myog, (B) Myod1 and (C) Myf5 determined by RT-qPCR normalized to the geometric mean of the housekeeping genes Actb, Tbp, Rplp0 and Rpl10. Similarly, the same tissues were analysed for expression of (D) miR-1, (E) miR-133a and (F) miR-206 by small RNA TaqMan RT-qPCR normalized to miR-16 expression. All values are mean + SEM, n = 3–4. (G) Correlation matrix comparing the pattern of expression across all comparable time points of all myogenic transcription factors, dystromiRs and novel miRNA biomarkers between in the TA, diaphragm and in serum. Scale bars indicate Spearman correlation coefficients. Red indicates a positive correlation, blue indicates a negative correlation and white indicates no correlation.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3814379&req=5

gkt724-F4: Serum dystromiR abundance correlates with expression of regeneration factors in the TA. TA muscles from mdx and Pip6e-PMO-treated mdx mice were harvested at various time points, and the expression of the myogenic transcription factors (A) Myog, (B) Myod1 and (C) Myf5 determined by RT-qPCR normalized to the geometric mean of the housekeeping genes Actb, Tbp, Rplp0 and Rpl10. Similarly, the same tissues were analysed for expression of (D) miR-1, (E) miR-133a and (F) miR-206 by small RNA TaqMan RT-qPCR normalized to miR-16 expression. All values are mean + SEM, n = 3–4. (G) Correlation matrix comparing the pattern of expression across all comparable time points of all myogenic transcription factors, dystromiRs and novel miRNA biomarkers between in the TA, diaphragm and in serum. Scale bars indicate Spearman correlation coefficients. Red indicates a positive correlation, blue indicates a negative correlation and white indicates no correlation.

Mentions: Given that both the previously described dystromiRs and the novel candidate miRNA biomarkers identified here showed complex patterns of serum abundance over time, which did not follow the pattern of dystrophin restoration, we reasoned that these miRNAs might reflect underlying pathophysiological processes occurring in muscle. As the dystromiRs of interest have established roles in the control of muscle development, we measured the expression of the myogenic transcription factors myogenin (Myog), Myod1 and Myf5 and the dystromiRs themselves by RT-qPCR in the TA (Figure 4A–F) and diaphragm (Supplementary Figure S3A–F) muscles, harvested from the same mice used in the time course study. To investigate the relationship between patterns of expression in the tissues and in serum, correlation analyses were performed between mRNAs/miRNAs over all comparable time points (Figure 4G, Supplementary Table S1).Figure 4.


Extracellular microRNAs are dynamic non-vesicular biomarkers of muscle turnover.

Roberts TC, Godfrey C, McClorey G, Vader P, Briggs D, Gardiner C, Aoki Y, Sargent I, Morgan JE, Wood MJ - Nucleic Acids Res. (2013)

Serum dystromiR abundance correlates with expression of regeneration factors in the TA. TA muscles from mdx and Pip6e-PMO-treated mdx mice were harvested at various time points, and the expression of the myogenic transcription factors (A) Myog, (B) Myod1 and (C) Myf5 determined by RT-qPCR normalized to the geometric mean of the housekeeping genes Actb, Tbp, Rplp0 and Rpl10. Similarly, the same tissues were analysed for expression of (D) miR-1, (E) miR-133a and (F) miR-206 by small RNA TaqMan RT-qPCR normalized to miR-16 expression. All values are mean + SEM, n = 3–4. (G) Correlation matrix comparing the pattern of expression across all comparable time points of all myogenic transcription factors, dystromiRs and novel miRNA biomarkers between in the TA, diaphragm and in serum. Scale bars indicate Spearman correlation coefficients. Red indicates a positive correlation, blue indicates a negative correlation and white indicates no correlation.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3814379&req=5

gkt724-F4: Serum dystromiR abundance correlates with expression of regeneration factors in the TA. TA muscles from mdx and Pip6e-PMO-treated mdx mice were harvested at various time points, and the expression of the myogenic transcription factors (A) Myog, (B) Myod1 and (C) Myf5 determined by RT-qPCR normalized to the geometric mean of the housekeeping genes Actb, Tbp, Rplp0 and Rpl10. Similarly, the same tissues were analysed for expression of (D) miR-1, (E) miR-133a and (F) miR-206 by small RNA TaqMan RT-qPCR normalized to miR-16 expression. All values are mean + SEM, n = 3–4. (G) Correlation matrix comparing the pattern of expression across all comparable time points of all myogenic transcription factors, dystromiRs and novel miRNA biomarkers between in the TA, diaphragm and in serum. Scale bars indicate Spearman correlation coefficients. Red indicates a positive correlation, blue indicates a negative correlation and white indicates no correlation.
Mentions: Given that both the previously described dystromiRs and the novel candidate miRNA biomarkers identified here showed complex patterns of serum abundance over time, which did not follow the pattern of dystrophin restoration, we reasoned that these miRNAs might reflect underlying pathophysiological processes occurring in muscle. As the dystromiRs of interest have established roles in the control of muscle development, we measured the expression of the myogenic transcription factors myogenin (Myog), Myod1 and Myf5 and the dystromiRs themselves by RT-qPCR in the TA (Figure 4A–F) and diaphragm (Supplementary Figure S3A–F) muscles, harvested from the same mice used in the time course study. To investigate the relationship between patterns of expression in the tissues and in serum, correlation analyses were performed between mRNAs/miRNAs over all comparable time points (Figure 4G, Supplementary Table S1).Figure 4.

Bottom Line: Expression of the myogenic miRNA, miR-206 and the myogenic transcription factor myogenin in the tibialis anterior muscle were found to positively correlate with serum dystromiR levels, suggesting that extracellular miRNAs are indicators of the regenerative status of the musculature.Only a minority of serum dystromiRs were found in extracellular vesicles, whereas the majority were protected from serum nucleases by association with protein/lipoprotein complexes.In conclusion, extracellular miRNAs are dynamic indices of pathophysiological processes in skeletal muscle.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford, OX1 3QX, UK, Department of Molecular and Experimental Medicine, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037, USA, Dubowitz Neuromuscular Centre, UCL Institute of Child Health, 30 Guildford Street, London, WC1N 1EH, UK and Nuffield Department of Obstetrics and Gynaecology, University of Oxford, Level 3, Women's Centre, John Radcliffe Hospital, Oxford, OX3 9DU, UK.

ABSTRACT
Extracellular microRNAs (miRNAs) are promising biomarkers of the inherited muscle wasting condition Duchenne muscular dystrophy, as they allow non-invasive monitoring of either disease progression or response to therapy. In this study, serum miRNA profiling reveals a distinct extracellular miRNA signature in dystrophin-deficient mdx mice, which shows profound dose-responsive restoration following dystrophin rescue. Extracellular dystrophy-associated miRNAs (dystromiRs) show dynamic patterns of expression that mirror the progression of muscle pathology in mdx mice. Expression of the myogenic miRNA, miR-206 and the myogenic transcription factor myogenin in the tibialis anterior muscle were found to positively correlate with serum dystromiR levels, suggesting that extracellular miRNAs are indicators of the regenerative status of the musculature. Similarly, extracellular dystromiRs were elevated following experimentally-induced skeletal muscle injury and regeneration in non-dystrophic mice. Only a minority of serum dystromiRs were found in extracellular vesicles, whereas the majority were protected from serum nucleases by association with protein/lipoprotein complexes. In conclusion, extracellular miRNAs are dynamic indices of pathophysiological processes in skeletal muscle.

Show MeSH
Related in: MedlinePlus