DNA methylation analysis of negative pressure therapy effect in diabetic foot ulcers

Abstract

ObjectiveNegative pressure wound therapy (NPWT) has been used to treat diabetic foot ulcerations (DFUs). Its action on the molecular level, however, is only partially understood. Some earlier data suggested NPWT may be mediated through modification of local gene expression. As methylation is a key epigenetic regulatory mechanism of gene expression, we assessed the effect of NPWT on its profile in patients with type 2 diabetes (T2DM) and neuropathic non-infected DFUs.MethodsOf 36 included patients, 23 were assigned to NPWT and 13 to standard therapy. Due to ethical concerns, the assignment was non-randomized and based on wound characteristics. Tissue samples were obtained before and 8 ± 1 days after therapy initiation. DNA methylation patterns were checked by Illumina Methylation EPIC kit.ResultsIn terms of clinical characteristics, the groups presented typical features of T2DM; however, the NPWT group had significantly greater wound area: 16.8 cm2 vs 1.4 cm2 (P = 0.0003). Initially only one region at chromosome 5 was differentially methylated. After treatment, 57 differentially methylated genes were found, mainly located on chromosomes 6 (chr6p21) and 20 (chr20p13); they were associated with DNA repair and autocrine signaling via retinoic acid receptor. We performed differential analyses pre treatment and post treatment. The analysis revealed 426 differentially methylated regions in the NPWT group, but none in the control group. The enrichment analysis showed 11 processes significantly associated with NPWT, of which 4 were linked with complement system activation. All but one were hypermethylated after NPWT.ConclusionThe NPWT effect on DFUs may be mediated through epigenetic changes resulting in the inhibition of complement system activation.

Negative pressure promotes macrophage M1 polarization after Mycobacterium tuberculosis infection via the lncRNA XIST/microRNA–125b–5p/A20/NF–κB axis

“…We found that negative pressure treatment downregulated XIST in MTB‐infected RAW264.7 cells and hMDMs by regulation of XIST promoter methylation. Previous work showed an association between negative pressure treatment and DNA methylation in diabetic foot ulcers 39 . We have now revealed that negative pressure treatment activates the NF‐κB pathway by downregulation of A20, which is miR‐125b‐5p‐targeted, through decreasing XIST expression.…”

“…Previous work showed an association between negative pressure treatment and DNA methylation in diabetic foot ulcers. 39 We have now revealed that negative pressure treatment activates the NF-κB pathway by downregulation of A20, which is miR-125b-5p-targeted, through decreasing XIST expression. The critical regulatory role of negative pressure wound therapy on plasma miRNA expression has been highlighted recently.…”

Effects of Negative Pressure Wound Therapy on Levels of Angiopoetin-2 and Other Selected Circulating Signaling Molecules in Patients with Diabetic Foot Ulcer

“…In our previous reports, we saw that NPWT altered the local gene expression involved with wound healing [4]. Moreover, we showed that NPWT's action is mediated through epigenetic alterations resulting mainly in the inhibition of complement system activation [5]. In a very recent study, NPWT increased the number of circulating endothelial progenitor cells in ischemic foot ulcers of diabetic patients which was attributed to the upregulation of systemic and local vascular endothelial growth factors and stromal cell-derived factor-1 α levels [28].…”

“…Negative pressure wound therapy (NPWT) has been used as an adjunct treatment for DFUs. Its potential influence on local gene expression and epigenetic methylation in the wound bed has been recently described [4, 5]. However, NPWT's novel mechanism of action may be associated with processes involving the entire organism where signals are transmitted via circulating molecules between the organism's tissues and organs.…”

A review of epigenetic regulation in wound healing: Implications for the future of wound care

“…When applied to wounds, closure is significantly accelerated, an effect that appears to be regenerative in nature through the activation of a plethora of neurodevelopmental genes 105 . Besides pharmacological modulation, even the application of negative pressure can induce local changes in wound bed methylation thereby activating the wound healing response 107 . Could methylation therefore be a useful target in the wound healing armamentarium?…”