miR-200c inhibition and catalase accelerate diabetic wound healing
Marco D'Agostino1, Sara Sileno1, Daniela Lulli2
1Molecular Regenerative Medicine Laboratory, Istituto Dermopatico dell'Immacolata (IDI-IRCCS), Rome, Italy.
Background:
Reactive oxygen species (ROS) are increased in diabetic conditions and play a causal role in diabetic foot ulcers (DFU). We previously showed that ROS up-regulate miR-200c expression, that in turns causes apoptosis, senescence, ROS upregulation and nitric oxide decrease, leading to endothelial disfunction.
Methods:
The aim of this study is to dissect miR-200c role in DFU and to explore the potential role of anti-miR-200c and antioxidant catalase (CAT) in promoting wound healing (WH). miR-200c inhibition and CAT treatment were performed either in immortalized keratinocytes (HaCaT) or in primary fibroblasts (FBs) and keratinocytes (KCs) deriving from diabetic patients (pts) undergoing amputations. Primary cells deriving from pts undergoing saphenectomies were used as controls. The miR-200c blockade was performed either via lentiviral particles bearing an anti-miR-200c sequence or locked nucleic acid (LNA) anti-miR-200c oligos. Equine CAT was administered on cell medium. The WH assay was performed in vivo on diabetic (db/db) mice by a topical treatment with CAT and LNA anti-miR-200c on wounds dissolved in a Pluronic gel mixture, administered every three days.
Results:
We found that miR-200c levels were increased by different stimuli known to induce ROS, such as ultraviolet radiation (UV), hydrogen peroxide (H2O2), and high glucose in HaCaT. miR-200c was also upregulated in skin biopsies, in FBs and KCs isolated from pts with DFU vs controls. Forced miR-200c expression induced ROS in both FBs and KCs, and CAT reduced it. miR-200c inhibition improved WH in HaCaT, both under basal conditions and after UV and H2O2 treatment, and the simultaneous treatment with CAT accelerated it. miR-200c inhibition accelerated WH in KCs of DFU pts, increasing its protein targets: sirtuin 1 (SIRT1), the transcription factors FOXO1 and ZEB1 and decreasing p66Shc phosphorylation at Ser-36, that is induced by ROS, and the co-treatment with CAT showed synergistic effects in reducing ROS and cytotoxicity. Interestingly, CAT treatment decreased miR-200c expression in FBs and KCs of DFU pts. Topical administration of anti-miR-200c and CAT in a WH model of diabetic mice accelerated closure.
Conclusions:
Anti-miR-200c and CAT could be considered a novel treatment for DFU and, possibly, for other types of non-diabetic skin ulcers.
Insights
Targeting miR-200c and using antioxidant catalase (CAT) shows promise for treating diabetic foot ulcers (DFU). This approach enhances wound healing (WH) by reducing reactive oxygen species (ROS) and improving cellular function.
Area of Science:
- Biomedical Science
- Molecular Biology
- Dermatology
Background:
- Diabetic conditions elevate reactive oxygen species (ROS), contributing to diabetic foot ulcers (DFU).
- Elevated ROS upregulate miR-200c, leading to apoptosis, senescence, and endothelial dysfunction.
- Previous research indicated a causal role for ROS in DFU pathogenesis.
Purpose of the Study:
- To investigate the role of miR-200c in diabetic foot ulcer (DFU) development.
- To evaluate the therapeutic potential of anti-miR-200c and antioxidant catalase (CAT) for wound healing (WH).
Main Methods:
- miR-200c inhibition and CAT treatment were applied to human keratinocytes and fibroblasts from diabetic patients and controls.
- In vivo studies utilized a diabetic mouse model (db/db) with topical application of anti-miR-200c and CAT.
- Wound healing assays were conducted using lentiviral particles, locked nucleic acid (LNA) oligos, and equine CAT.
Main Results:
- miR-200c levels were increased by ROS-inducing stimuli and were higher in DFU patient cells.
- miR-200c inhibition and CAT treatment reduced ROS, improved wound healing in vitro and in vivo.
- Co-treatment with anti-miR-200c and CAT demonstrated synergistic effects in reducing ROS and cytotoxicity.
Conclusions:
- Anti-miR-200c and catalase represent a potential novel therapeutic strategy for diabetic foot ulcers (DFU).
- This combined approach may also be beneficial for other non-diabetic skin ulcer types.
- Targeting miR-200c and ROS offers a promising avenue for enhancing wound healing.
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