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RNA sequencing reveals a depletion of collagen targeting microRNAs in Dupuytren's disease.

Riester SM, Arsoy D, Camilleri ET, Dudakovic A, Paradise CR, Evans JM, Torres-Mora J, Rizzo M, Kloen P, Julio MK, van Wijnen AJ, Kakar S - BMC Med Genomics (2015)

Bottom Line: RT-qPCR confirmed the decreased expression of microRNA targeted collagens in control palmar fascia tissues.Control palmar fascia show decreased expression of mRNAs encoding collagens that are preferentially targeted by microRNAs enriched in non-diseased fascia.Dupuytren's fascia and healthy palmar fascia can be distinguished by unique microRNA profiles, which are predicted to preferentially target collagens and other extracellular matrix proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, MN, 55905, USA. riester.scott@mayo.edu.

ABSTRACT

Background: Dupuytren's disease is an inherited disorder in which patients develop fibrotic contractures of the hand. Current treatment strategies include surgical excision or enzymatic digestion of fibrotic tissue. MicroRNAs, which are key posttranscriptional regulators of genes expression, have been shown to play an important regulatory role in disorders of fibrosis. Therefore in this investigation, we apply high throughput next generation RNA sequencing strategies to characterize microRNA expression in diseased and healthy palmar fascia to elucidate molecular mechanisms responsible for pathogenic fibrosis.

Methods: We applied high throughput RNA sequencing techniques to quantify the expression of all known human microRNAs in Dupuytren's and control palmar fascia. MicroRNAs that were differentially expressed between diseased and healthy tissue samples were used for computational target prediction using the bioinformatics tool ComiR. Molecular pathways that were predicted to be differentially expressed based on computational analysis were validated by performing RT-qPCR on RNA extracted from diseased and non-diseased palmar fascia biopsies.

Results: A comparison of microRNAs expressed in Dupuytren's fascia and control fascia identified 74 microRNAs with a 2-fold enrichment in Dupuytren's tissue, and 32 microRNAs with enrichment in control fascia. Computational target prediction for differentially expressed microRNAs indicated preferential targeting of collagens and extracellular matrix related proteins in control palmar fascia. RT-qPCR confirmed the decreased expression of microRNA targeted collagens in control palmar fascia tissues.

Discussion: Control palmar fascia show decreased expression of mRNAs encoding collagens that are preferentially targeted by microRNAs enriched in non-diseased fascia. Thus alterations in microRNA regulatory networks may play an important role in driving the pathogenic fibrosis seen in Dupuytren's disease via direct regulatory effects on extracellular matrix protein synthesis.

Conclusion: Dupuytren's fascia and healthy palmar fascia can be distinguished by unique microRNA profiles, which are predicted to preferentially target collagens and other extracellular matrix proteins.

No MeSH data available.


Related in: MedlinePlus

A RT-qPCR screen for fibrosis associated mRNAs linked to Wnt and TGFβ signaling are shown. This analysis reveals a statistically significant upregulation of WNT5A in Dupuytren’s fascia, and a paradoxical upregulation of TGFBR2 in control fascia. WNT5A is predicted to be downregulated by microRNAs enriched in Dupuytren’s disease based on ComiR analysis, thus WNT5A upregulation is unlikely to be mediated solely by microRNA regulatory. In accordance with ComiR target predictions, we do not observe strong evidence for direct microRNA regulation of TGFβ and Wnt signaling pathways in Dupuytren’s disease
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Fig8: A RT-qPCR screen for fibrosis associated mRNAs linked to Wnt and TGFβ signaling are shown. This analysis reveals a statistically significant upregulation of WNT5A in Dupuytren’s fascia, and a paradoxical upregulation of TGFBR2 in control fascia. WNT5A is predicted to be downregulated by microRNAs enriched in Dupuytren’s disease based on ComiR analysis, thus WNT5A upregulation is unlikely to be mediated solely by microRNA regulatory. In accordance with ComiR target predictions, we do not observe strong evidence for direct microRNA regulation of TGFβ and Wnt signaling pathways in Dupuytren’s disease

Mentions: Previous studies have implicated alterations in Wnt and TGFβ signaling in the pathogenesis of Dupuytren’s disease [7, 74, 75]. To determine if microRNAs regulate either of these pathways, we evaluated the expression of mRNAs associated with these two signaling pathways using RT-qPCR (Fig. 8). TGFBR2 and WNT5A were the only two genes to show statistically significant differences in expression between the Dupuytren’s and control palmar fascia. TGFBR2 paradoxically showed increased expression in the control fascia, enrichment inconsistent with its known fibrosis promoting affects. WNT5A was the only gene to show a statistically significant enrichment in the Dupuytren’s fascia. However microRNA target prediction indicates that WNT5A is preferentially inhibited by microRNAs enriched in Dupuytren’s fascia, indicating that microRNA regulation is unlikely to be driving the increased expression of WNT5A in diseased palmar fascia. These findings suggest that microRNAs, which promote mRNA degradation, do not play a major role in post-transcriptional regulation of WNT and TGFβ signaling pathways in the terminal stages of Dupuytren’s disease.Fig. 8


RNA sequencing reveals a depletion of collagen targeting microRNAs in Dupuytren's disease.

Riester SM, Arsoy D, Camilleri ET, Dudakovic A, Paradise CR, Evans JM, Torres-Mora J, Rizzo M, Kloen P, Julio MK, van Wijnen AJ, Kakar S - BMC Med Genomics (2015)

A RT-qPCR screen for fibrosis associated mRNAs linked to Wnt and TGFβ signaling are shown. This analysis reveals a statistically significant upregulation of WNT5A in Dupuytren’s fascia, and a paradoxical upregulation of TGFBR2 in control fascia. WNT5A is predicted to be downregulated by microRNAs enriched in Dupuytren’s disease based on ComiR analysis, thus WNT5A upregulation is unlikely to be mediated solely by microRNA regulatory. In accordance with ComiR target predictions, we do not observe strong evidence for direct microRNA regulation of TGFβ and Wnt signaling pathways in Dupuytren’s disease
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4597401&req=5

Fig8: A RT-qPCR screen for fibrosis associated mRNAs linked to Wnt and TGFβ signaling are shown. This analysis reveals a statistically significant upregulation of WNT5A in Dupuytren’s fascia, and a paradoxical upregulation of TGFBR2 in control fascia. WNT5A is predicted to be downregulated by microRNAs enriched in Dupuytren’s disease based on ComiR analysis, thus WNT5A upregulation is unlikely to be mediated solely by microRNA regulatory. In accordance with ComiR target predictions, we do not observe strong evidence for direct microRNA regulation of TGFβ and Wnt signaling pathways in Dupuytren’s disease
Mentions: Previous studies have implicated alterations in Wnt and TGFβ signaling in the pathogenesis of Dupuytren’s disease [7, 74, 75]. To determine if microRNAs regulate either of these pathways, we evaluated the expression of mRNAs associated with these two signaling pathways using RT-qPCR (Fig. 8). TGFBR2 and WNT5A were the only two genes to show statistically significant differences in expression between the Dupuytren’s and control palmar fascia. TGFBR2 paradoxically showed increased expression in the control fascia, enrichment inconsistent with its known fibrosis promoting affects. WNT5A was the only gene to show a statistically significant enrichment in the Dupuytren’s fascia. However microRNA target prediction indicates that WNT5A is preferentially inhibited by microRNAs enriched in Dupuytren’s fascia, indicating that microRNA regulation is unlikely to be driving the increased expression of WNT5A in diseased palmar fascia. These findings suggest that microRNAs, which promote mRNA degradation, do not play a major role in post-transcriptional regulation of WNT and TGFβ signaling pathways in the terminal stages of Dupuytren’s disease.Fig. 8

Bottom Line: RT-qPCR confirmed the decreased expression of microRNA targeted collagens in control palmar fascia tissues.Control palmar fascia show decreased expression of mRNAs encoding collagens that are preferentially targeted by microRNAs enriched in non-diseased fascia.Dupuytren's fascia and healthy palmar fascia can be distinguished by unique microRNA profiles, which are predicted to preferentially target collagens and other extracellular matrix proteins.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, MN, 55905, USA. riester.scott@mayo.edu.

ABSTRACT

Background: Dupuytren's disease is an inherited disorder in which patients develop fibrotic contractures of the hand. Current treatment strategies include surgical excision or enzymatic digestion of fibrotic tissue. MicroRNAs, which are key posttranscriptional regulators of genes expression, have been shown to play an important regulatory role in disorders of fibrosis. Therefore in this investigation, we apply high throughput next generation RNA sequencing strategies to characterize microRNA expression in diseased and healthy palmar fascia to elucidate molecular mechanisms responsible for pathogenic fibrosis.

Methods: We applied high throughput RNA sequencing techniques to quantify the expression of all known human microRNAs in Dupuytren's and control palmar fascia. MicroRNAs that were differentially expressed between diseased and healthy tissue samples were used for computational target prediction using the bioinformatics tool ComiR. Molecular pathways that were predicted to be differentially expressed based on computational analysis were validated by performing RT-qPCR on RNA extracted from diseased and non-diseased palmar fascia biopsies.

Results: A comparison of microRNAs expressed in Dupuytren's fascia and control fascia identified 74 microRNAs with a 2-fold enrichment in Dupuytren's tissue, and 32 microRNAs with enrichment in control fascia. Computational target prediction for differentially expressed microRNAs indicated preferential targeting of collagens and extracellular matrix related proteins in control palmar fascia. RT-qPCR confirmed the decreased expression of microRNA targeted collagens in control palmar fascia tissues.

Discussion: Control palmar fascia show decreased expression of mRNAs encoding collagens that are preferentially targeted by microRNAs enriched in non-diseased fascia. Thus alterations in microRNA regulatory networks may play an important role in driving the pathogenic fibrosis seen in Dupuytren's disease via direct regulatory effects on extracellular matrix protein synthesis.

Conclusion: Dupuytren's fascia and healthy palmar fascia can be distinguished by unique microRNA profiles, which are predicted to preferentially target collagens and other extracellular matrix proteins.

No MeSH data available.


Related in: MedlinePlus