Limits...
MicroRNA 181b regulates decorin production by dermal fibroblasts and may be a potential therapy for hypertrophic scar.

Kwan P, Ding J, Tredget EE - PLoS ONE (2015)

Bottom Line: Lower decorin levels were found in hypertrophic scar as compared to normal skin, and in deep as compared to superficial dermis.By blocking miR-181b with an antagomiR, it was possible to increase decorin protein expression in dermal fibroblasts.This suggests miR-181b is involved in the differential expression of decorin in skin and wound healing.

View Article: PubMed Central - PubMed

Affiliation: Division of Plastic Surgery, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada; Wound Healing Research Group, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.

ABSTRACT
Hypertrophic scarring is a frequent fibroproliferative complication following deep dermal burns leading to impaired function and lifelong disfigurement. Decorin reduces fibrosis and induces regeneration in many tissues, and is significantly downregulated in hypertrophic scar and normal deep dermal fibroblasts. It was hypothesized that microRNAs in these fibroblasts downregulate decorin and blocking them would increase decorin and may prevent hypertrophic scarring. Lower decorin levels were found in hypertrophic scar as compared to normal skin, and in deep as compared to superficial dermis. A decorin 3' un-translated region reporter assay demonstrated microRNA decreased decorin in deep dermal fibroblasts, and microRNA screening predicted miR- 24, 181b, 421, 526b, or 543 as candidates. After finding increased levels of mir-181b in deep dermal fibroblasts, it was demonstrated that TGF-β1 stimulation decreased miR-24 but increased miR-181b and that hypertrophic scar and deep dermis contained increased levels of miR-181b. By blocking miR-181b with an antagomiR, it was possible to increase decorin protein expression in dermal fibroblasts. This suggests miR-181b is involved in the differential expression of decorin in skin and wound healing. Furthermore, blocking miR-181b reversed TGF-β1 induced decorin downregulation and myofibroblast differentiation in hypertrophic scar fibroblasts, suggesting a potential therapy for hypertrophic scar.

No MeSH data available.


Related in: MedlinePlus

Regulation of miRNA expression by TGF-β1 in SF and DF.Cells were cultured in DMEM + 2% FBS with the indicated treatment protocols and total RNA extracted for RT-qPCR. (a) Dose-response curve showing relative expression of miR-24 for SF and DF cultured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3). (b) Dose-response curve showing relative expression of miR-181b for SF and DF culutured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3, * P < 0.05, ** P < 0.01). (c) Time-response curve showing relative expression of miR-181b for SF and DF at fixed concentrations of TGF-β1 (SF 10 ng/mL, DF 20 ng/mL) (mean ± SEM, n = 3, * P < 0.03).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4383602&req=5

pone.0123054.g003: Regulation of miRNA expression by TGF-β1 in SF and DF.Cells were cultured in DMEM + 2% FBS with the indicated treatment protocols and total RNA extracted for RT-qPCR. (a) Dose-response curve showing relative expression of miR-24 for SF and DF cultured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3). (b) Dose-response curve showing relative expression of miR-181b for SF and DF culutured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3, * P < 0.05, ** P < 0.01). (c) Time-response curve showing relative expression of miR-181b for SF and DF at fixed concentrations of TGF-β1 (SF 10 ng/mL, DF 20 ng/mL) (mean ± SEM, n = 3, * P < 0.03).

Mentions: Since TGF-β1 is a key profibrotic cytokine in HSc development [44], its effects on miR-24 and miR-181b in SF and DF were examined using RT-qPCR. miR-24 was downregulated by TGF-β1 in SF and DF in a dose-dependent manner (Fig 3A), in keeping with findings in myoblasts [45]. In contrast, miR-181b was significantly upregulated by a low concentration of TGF-β1 in SF (10 ng/mL) and DF (20 ng/mL) with a return to baseline by a high concentration of TGF-β1 (40 ng/mL) (Fig 3B), and the upregulation of miRNA-181b by TGF-β1 in SF (10 ng/mL) and DF (20 ng/mL) was a time-dependent manner (Fig 3C), similar to observations in hepatocytes [46]. A similar experiment using CTGF stimulation did not show changes in miR-181b expression. Based on these results miR-181b was selected for further investigation.


MicroRNA 181b regulates decorin production by dermal fibroblasts and may be a potential therapy for hypertrophic scar.

Kwan P, Ding J, Tredget EE - PLoS ONE (2015)

Regulation of miRNA expression by TGF-β1 in SF and DF.Cells were cultured in DMEM + 2% FBS with the indicated treatment protocols and total RNA extracted for RT-qPCR. (a) Dose-response curve showing relative expression of miR-24 for SF and DF cultured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3). (b) Dose-response curve showing relative expression of miR-181b for SF and DF culutured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3, * P < 0.05, ** P < 0.01). (c) Time-response curve showing relative expression of miR-181b for SF and DF at fixed concentrations of TGF-β1 (SF 10 ng/mL, DF 20 ng/mL) (mean ± SEM, n = 3, * P < 0.03).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383602&req=5

pone.0123054.g003: Regulation of miRNA expression by TGF-β1 in SF and DF.Cells were cultured in DMEM + 2% FBS with the indicated treatment protocols and total RNA extracted for RT-qPCR. (a) Dose-response curve showing relative expression of miR-24 for SF and DF cultured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3). (b) Dose-response curve showing relative expression of miR-181b for SF and DF culutured in increasing concentrations of TGF-β1 for 48 hours (mean ± SEM, n = 3, * P < 0.05, ** P < 0.01). (c) Time-response curve showing relative expression of miR-181b for SF and DF at fixed concentrations of TGF-β1 (SF 10 ng/mL, DF 20 ng/mL) (mean ± SEM, n = 3, * P < 0.03).
Mentions: Since TGF-β1 is a key profibrotic cytokine in HSc development [44], its effects on miR-24 and miR-181b in SF and DF were examined using RT-qPCR. miR-24 was downregulated by TGF-β1 in SF and DF in a dose-dependent manner (Fig 3A), in keeping with findings in myoblasts [45]. In contrast, miR-181b was significantly upregulated by a low concentration of TGF-β1 in SF (10 ng/mL) and DF (20 ng/mL) with a return to baseline by a high concentration of TGF-β1 (40 ng/mL) (Fig 3B), and the upregulation of miRNA-181b by TGF-β1 in SF (10 ng/mL) and DF (20 ng/mL) was a time-dependent manner (Fig 3C), similar to observations in hepatocytes [46]. A similar experiment using CTGF stimulation did not show changes in miR-181b expression. Based on these results miR-181b was selected for further investigation.

Bottom Line: Lower decorin levels were found in hypertrophic scar as compared to normal skin, and in deep as compared to superficial dermis.By blocking miR-181b with an antagomiR, it was possible to increase decorin protein expression in dermal fibroblasts.This suggests miR-181b is involved in the differential expression of decorin in skin and wound healing.

View Article: PubMed Central - PubMed

Affiliation: Division of Plastic Surgery, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada; Wound Healing Research Group, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.

ABSTRACT
Hypertrophic scarring is a frequent fibroproliferative complication following deep dermal burns leading to impaired function and lifelong disfigurement. Decorin reduces fibrosis and induces regeneration in many tissues, and is significantly downregulated in hypertrophic scar and normal deep dermal fibroblasts. It was hypothesized that microRNAs in these fibroblasts downregulate decorin and blocking them would increase decorin and may prevent hypertrophic scarring. Lower decorin levels were found in hypertrophic scar as compared to normal skin, and in deep as compared to superficial dermis. A decorin 3' un-translated region reporter assay demonstrated microRNA decreased decorin in deep dermal fibroblasts, and microRNA screening predicted miR- 24, 181b, 421, 526b, or 543 as candidates. After finding increased levels of mir-181b in deep dermal fibroblasts, it was demonstrated that TGF-β1 stimulation decreased miR-24 but increased miR-181b and that hypertrophic scar and deep dermis contained increased levels of miR-181b. By blocking miR-181b with an antagomiR, it was possible to increase decorin protein expression in dermal fibroblasts. This suggests miR-181b is involved in the differential expression of decorin in skin and wound healing. Furthermore, blocking miR-181b reversed TGF-β1 induced decorin downregulation and myofibroblast differentiation in hypertrophic scar fibroblasts, suggesting a potential therapy for hypertrophic scar.

No MeSH data available.


Related in: MedlinePlus