Unknown,Transcriptomics,Genomics,Proteomics

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Gene Expression Profiles in Thermally Injured Human Skin: A Temporal Microarray Analysis

ABSTRACT: Thermal injury incites inflammatory responses that often transcend the local environment and lead to structural deficiencies in skin that give way to scar formation. We hypothesized that extensive perturbations within burned skin following thermal insult and during subsequent events of wound repair induce vast alterations in gene expression that likely serve as a wound and systemic healing deterrent. A high-throughput microarray experiment was designed to analyze genetic expression patterns and identify potential genes to target for therapeutic augmentation or silencing. The study compares gene expression from burn wound margins at various times following thermal injury to expression observed in normal skin. Utilizing this design, we report that the totality of gene expression alterations is indeed enormous. Further, we observed that the differential expression of many inflammatory and immune response genes appear to be continually up-regulated in burn wound margins seven days or more after initial thermal insult. As it is well established that the inflammatory process must abate for wound healing to proceed, the finding of ongoing local inflammation is cause for further investigation. To our knowledge, this is the first report of the gene expression alterations induced by thermal injury of human skin. As such, it provides a wealth of data to mine with the ultimate goal of better understanding the local pathophysiologic changes at the site of thermal injury that not only affect wound healing capacity, but may also contribute to systemic derangements within the burn patient. Keywords: time course, disease state analysis The study compares gene expression from burn wound margins at various times following thermal injury to expression observed in normal skin. All skin specimens were obtained in the operating room within minutes of being removed from the patient. Burn specimens were taken from wound margins. Harvested tissue at the burn wound margin maximized the capture of viable cells from the multiple lineages important to the healing process and minimized the inclusion of non-viable cells destroyed by full-thickness injury. After isolation, RNA samples were pooled equally by mass as to contain RNA from 5 tissue specimens for each array replicate.

ORGANISM(S): Homo sapiens

SUBMITTER: Lillian Nanney

PROVIDER: E-GEOD-8056 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Negative pressure wound therapy (NPWT) has been used to promote wound healing in a variety of settings, including as an adjunct to silver-impregnated dressings in the acute management of paediatric burns. Fluid aspirated by the NPWT system represents a potentially insightful research matrix for understanding the burn wound microenvironment and the biochemical mechanisms of action of NPWT. The aim of this study was to characterise the proteome of wound fluid collected using NPWT from children with small-area thermal burns. Samples were obtained as part of a randomised controlled trial investigating the clinical efficacy of adjunctive NPWT. They were compared with blister fluid specimens from paediatric burn patients matched according to demographic and injury characteristics. Protein identification and quantification were performed via liquid chromatography tandem mass spectrometry (LC-MS/MS) and sequential window acquisition of all theoretical mass spectra (SWATH) data-independent acquisition. Proteins and biological pathways which were unique or enriched in negative pressure fluid samples were evaluated using principal components, partial least squares-discriminant, and gene ontology enrichment analyses. Eight viable samples of NPWT fluid were collected and analysed with eight matched blister fluid samples. A total of 502 proteins were quantitatively profiled in the NPWT fluid, of which 444 (88.4%) were shared with blister fluid. Several proteins exhibited significant abundance differences between fluid types, with NPWT fluid showing a higher abundance of proteases matrix metalloprotease-9 and arginase-1, immune cell surface molecule low affinity immunoglobulin gamma Fc region receptor III-A, actin-binding protein filamin-A, plasma protein alpha-2-macroglobulin, and haemoglobin subunit alpha. The results lend support to the hypothesis that NPWT augments wound healing through the modulation of factors involved in the inflammatory response, granulation tissue synthesis, and extracellular matrix maintenance.

Dataset Information

Gene Expression Profiles in Thermally Injured Human Skin: A Temporal Microarray Analysis

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