Project description:In this project, we analysed the proteome of skin wounds to investigate whether a vitamin A-deficient diet influences the wound proteome. To achieve this, we conducted quantitative, unbiased proteomic analysis using data-independent acquisition (DIA) mass spectrometr using skin wounds lysates.
Project description:In this analysis, we assessed the subtle changes in retinol metabolism during wound healing, by analyzing the proteome of normal and wounded skin in Vitamin A deficient -treated mice using targeted proteomics,analysis, SureQuant.
Project description:Diabetic foot ulcers remain a major clinical challenge as diabetes prevalence rises, emphasizing the need for improved therapeutics and relevant preclinical models. Common rodent wound healing models poorly recapitulate human skin anatomy and repair. Although porcine skin is comparable to human skin, many studies employ young, healthy pigs that do not reflect typical diabetic human wounds. Here, we evaluated wound healing in full-thickness skin wounds in non-diabetic and diabetic Yucatan minipigs. RNA sequencing identified key transcriptional differences in wounds of diabetic versus non-diabetic animals, including pathways linked to increased inflammation and oxidative stress, as well as decreased metabolism and extracellular matrix organization, known hallmarks of diabetic wounds. These findings support this preclinical model as a powerful approach for discovery and therapeutic testing in diabetic wounds and provide a novel data set for further mining of potential gene targets for diabetic wound intervention.
Project description:Fibroblasts are the primary cells responsible for depositing extracellular matrix proteins during wound repair and fibrosis. Progress towards the development of treatments aimed at reducing scars has been hampered by our limited understanding of specific fibroblast populations responsible for fibrotic versus regenerative healing. Using a four-site wounding model in mice, we observed that neural crest-derived facial wounds heal with less scarring than cephalic mesoderm-derived scalp wounds, lateral plate mesoderm-derived ventral wounds, and paraxial mesoderm-derived dorsal wounds. Single-cell RNA sequencing identified increased expression of Robo2 and downstream Eid1 in neural-crest derived facial fibroblasts compared to fibroblasts from other sites. Subsequent fibroblast transplantation experiments showed that Robo2 and Eid1 are required for facial fibroblasts’ intrinsic reduced fibrotic potential. This is maintained by the EID1-mediated inhibition of EP300 histone acetyltransferase, leading to a more transcriptionally silent chromatin landscape, including around extracellular matrix genes. Mimicking EID1’s endogenous activity, both small-molecule and transgenic repression of EP300 in dorsal wounds promoted facial-like healing with reduced scarring. Overall, these data highlight the importance of the ROBO2-EID1-EP300 signaling axis in facial wound healing, and demonstrate our ability to modulate the embryologically determined fibrogenic potential of fibroblasts as a therapeutic approach to minimize scar formation.
Project description:When compared to skin, oral mucosal wounds heal rapidly and with reduced scar formation. This study used an Affymetrix microarray platform to compare the transcriptomes of oral mucosa and skin wounds in order to identify critical differences in the healing response at these two sites. Using microarrays, we explored the differences in gene expression in skin and oral mucosal wound healing in a murine model of paired equivalent-sized wounds. Samples were examined from day 0 to day 10 and spanned all stages of the wound healing process. Unwounded matched tissue was used as a control. Tissue samples collected at each post-wounding time point, as well as control samples, were represented by 3 biological replicates.
Project description:In order to clarify the human response of re-epithelialization, we biopsied split-thickness skin graft donor site wounds immediately before and after harvesting, as well as during the healing process 3 and 7 days thereafter. Altogether 25 biopsies from 8 patients qualified for the study. All samples were analysed by genome-wide microarrays. Here we identified the genes associated with normal skin re-epithelialization on time-scale, and organized them by similarities according to their induction or suppression patterns during wound healing. Overall 25 samples were analyzed
Project description:Diabetic foot ulcer (DFU) is a common complication of diabetes characterized by increased inflammation and a slowed healing process for wounds. Interleukin-37 (IL-37) may act as an alarm to alert the immune system when released by epithelial barrier tissues during trauma or infection, exerting a broad range of protective effects in several diseases. The objective of this study was to examine the regenerative capabilities of IL-37 in improving the healing of diabetic wounds. Using streptozotocin (STZ)-induced diabetic mice, we found that diabetic IL-37Tg mice showed a significantly accelerated healing process. In addition, IL-37 strongly suppressed MAPK signaling pathway by inhibiting phosphorylation of the P38 and ERK. Moreover, IL-37 reduced the expression of Nod-like receptor protein-3 (NLRP3) and mature IL-1β. These results thus indicated that IL-37 inhibition of IL-1β production is mediated by suppressing the initial priming step and by inhibiting the NLRP3 inflammasome activation. Taken together, our findings demonstrated the promising regulatory activity of IL-37 against IL-1β production and indicated that IL-37 has the potential to be effective as a novel therapeutic agent for treatment of wound. Our data indicate a beneficial effect of IL-37 in diabetic wounds, suggesting a therapeutic potential for this cytokine in diabetic ulcer management.
Project description:Impaired skin wound healing is a significant global health issue, especially among the elderly. Wound healing is a well-orchestrated process involving the sequential phases of inflammation, proliferation, and tissue remodeling. Although wound healing is a highly dynamic and energy-requiring process, the role of metabolism remains largely unexplored. By combining transcriptomics and metabolomics of human skin biopsy samples, we mapped the core bioenergetic and metabolic changes in normal acute as well as chronic wounds in elderly subjects. We found upregulation of glycolysis, the tricarboxylic acid cycle, glutaminolysis, and β-oxidation in the later stages of acute wound healing and in chronic wounds. To ascertain the role of these metabolic pathways on wound healing, we targeted each pathway in a wound healing assay as well as in a human skin explant model using metabolic inhibitors and stimulants. Enhancement or inhibition of glycolysis and, to a lesser extent, glutaminolysis had a far greater impact on wound healing than similar manipulations of oxidative phosphorylation and fatty acid β-oxidation. These findings increase the understanding of wound metabolism and identify glycolysis and glutaminolysis as potential targets for therapeutic intervention.