Project description:Currently there is growing concern with respect to scenarios where people are likely to be presented with radiation exposure along with many kinds of other injuries such as trauma and infection. The potential for such scenarios was brought to reality with the events and aftermath of the Fukushima nuclear disaster in Japan. As such medical complications arising from such exposures would be poorly dealt with as no evidence-based guidelines exist for their rehabilitation or recovery. Our research intends to differentially characterize combined radiation and burn injuries and identify novel pathways and biomarkers. Such findings will lead to better medical practices in the diagnosis, care and rehabilitation of affected individuals. The study includes four groups of mice: 1) Control sham mice group (n=4), 2) Skin burn injury mice group (n=6), 3) Radiation injury mice group (n=6), 4) Combined radiation and burn injury mice group (n=6). We propose to characterize the effects of combined radiation and burn injuries using microRNA microarray analysis. Our primary aim is to identify novel molecular pathways and biomarkers specific to whole blood samples (serum) from mice exposed to combined radiation and burn injuries. B6D2F1/J female mice will be used. 30 days following combined radiation and burn injuries arterial blood will be harvested from euthanized mice. 200ul of serum from whole blood samples will be used for microRNA microarray experiments (Affymetrix).
Project description:Currently there is growing concern with respect to scenarios where people are likely to be presented with radiation exposure along with many kinds of other injuries such as trauma and infection. The potential for such scenarios was brought to reality with the events and aftermath of the Fukushima nuclear disaster in Japan. As such medical complications arising from such exposures would be poorly dealt with as no evidence-based guidelines exist for their rehabilitation or recovery. Our research intends to differentially characterize combined radiation and burn injuries and identify novel pathways and biomarkers. Such findings will lead to better medical practices in the diagnosis, care and rehabilitation of affected individuals. The study includes four groups of mice: 1) Control sham mice group (n=4), 2) Skin burn injury mice group (n=6), 3) Radiation injury mice group (n=6), 4) Combined radiation and burn injury mice group (n=6).
Project description:Full thickness and deep partial thickness burn injuries heal by scarring. There are several mechanisms thought to be essential for the development of burn scars, but a challenge to studying the skin response to burn injury is that there are few animal models of burn scarring that are either clinically similar to human burn scars or are practical for most investigators to use. The purpose of this study was to examine the changes in RNA expression in human skin to burn injury. This was done by comparing pre-injury tissue from otherwise healthy adults undergoing aesthetic scarification created by branding with a hot metal object to serial samples of untreated wounds in the same subjects.
Project description:Burn injuries are devastating traumas, often leading to life-long consequences that extend beyond the observable burn scar. Burn injury patients commonly develop chronic neurological disorders but the long-lasting impacts of burn injuries on neurons and glia in the brain is unknown. Whole transcriptome RNA-sequencing from cortical excitatory neurons, inhibitory neurons, astrocytes and microglia showed very few changes to the expression of genes with known functions five weeks following a non-severe burn injury in adult mice. However, genes related to GABA-A receptors in excitatory neurons and several cellular functions in microglia was found to be to differentially expressed in burn injured mice. These findings shed light on the long-term effect of burn injuries on the brain and may help identify potential therapeutic targets and windows to prevent neurological dysfunction in burn patients.
Project description:Investigation into murine dermal burn wound. Mouse thermal injury induced, and skin excised at 0 hours, 2 hours, 3 days and 14 days post-injury. Transcription profiling of skin excised from thermal injured mouse to investigate the molecular mechanism of murine dermal burn wound.
Project description:Keloid radiotherapy is clinically effective but has an incomplete underlying molecular mechanism and limited accessibility; we used X-ray exposure to identify effector pathways replicating its antifibrotic benefits without further radiation. After collecting patient keloid samples, primary human keloid fibroblasts and normal skin fibroblasts, non-targeted metabolomics and RNA-seq were performed, identifying phytosphingosine (PHS) and KIF20A as key mediators of fibrosis and cell death in keloid radiotherapy. X-ray irradiation increased PHS release in keloid fibroblasts; exogenous PHS and X-ray irradiation independently induced cell death and reduced fibrosis, their combination exerted radiosensitization by boosting G2/M arrest and apoptosis while lowering fibrosis progression, and both PHS and irradiation reduced KIF20A whose loss induced cell cycle arrest and apoptosis. Our findings reveal that X-ray irradiation enhances keloid fibroblast PHS secretion that downregulates KIF20A to exert keloid therapeutic effects, and topical PHS or KIF20A-targeting agents are immediately translatable alternatives for radiotherapy-ineligible patients.
Project description:The study was designed to determine the differential gene expression between burn eschar- and normal skin-derived pericytes. A comparison was also made to determine the gene expression between normal skin pericytes and normal skin fibroblasts and (2) comparison of differential gene expression between burn eschar pericytes and normal normal skin fibroblasts
Project description:Neutrophils accumulate early in burn wounds, and their activation is associated with more severe burns. Understanding their functionality would facilitate the development of a more targeted therapeutic strategy for healing. However, we still lack a view of the cellular and functional heterogeneity in neutrophils involved in thermal injuries. Here, we establish the use of larval zebrafish for understanding neutrophil responses in burn. Zebrafish model allows for linking neutrophil states and their functions in real time through genetic modifications and live imaging. We performed single-cell transcriptional (scRNA-Seq) mapping of myeloid cells during a 3-day time course in burn and unwounded conditions. We identified transcriptionally distinct states in myeloid cells that form a consistent population structure across time points and conditions. By comparing burn and unwounded conditions, we found subtype-specific enrichment of biological processes and differential usage of gene regulatory networks. Pseudotime and RNA velocity analyses predict distinct branched trajectories for neutrophils, with one branch resembling the process of human neutrophil maturation. The other branch is not transcriptionally conserved with humans and is highly associated with leukocyte migration functionality, suggesting its engagement at the wound site. Transcriptional network analysis identified RAR/RXR family transcription factors as potential upstream factors driving this trajectory divergence in neutrophils. Furthermore, we characterized the transcriptional dynamics of cell-cell interactions in both conditions by time. Among burn-induced signaling pathways, we found il6-il6r signaling as a time-point-specific macrophage-neutrophil interaction, suggesting the importance of timing in innate immune response in burn. Finally, to test the translational value of our fish model, we examined zebrafish neutrophil state signatures in human burn patient samples. We found homolog expression of immature neutrophils positively correlates with the degree of total body surface area (TBSA) in patients. Flow cytometry confirmed the presence of neutrophils carrying these signatures in patient's blood. Our findings demonstrate the potential of using zebrafish as a model to identify early innate immune response signatures that could inform a timely treatment of burn in humans. This work builds the molecular foundation and a comparative single-cell genomic framework to guide future identification of actionable pathways to burn wound healing in patients.
Project description:Severe burns are a major global health concern, and are associated with long-term physical and psychological impairments, multi-organ dysfunction, and substantial morbidity and mortality. While burn injuries in adults trigger systemic immuno-metabolic alterations—characterized by white adipose tissue browning, elevated resting energy expenditure, widespread catabolism and inflammation—these adaptive responses are considerably impaired in older adults, with molecular mechanisms behind these differences remaining largely unclear. As a key regulator of systemic metabolism, investigating the pathological role of adipose tissue (AT) post-burn may reveal novel targets that could potentially improve patient outcomes. In this study, we conducted bulk mRNA sequencing and analysis of AT from adult and aged mice, to elucidate the transcriptomic changes underlying the distinct post-burn responses in these populations. After examining differentially expressed genes in the adult and aged burn mice, the top six upregulated genes in adults (Ucp1, Lgr6, Dio2, Lncbate10, Fabp3, Kng2) were primarily associated with thermogenesis, whereas those in the aged mice (Car6, Spata25, Gm128, Btbd16, Lipm, Abca13) were linked to inflammation, tissue repair, and lipid metabolism. Furthermore, our gene co-expression and enrichment map analysis identified burn-associated modules related to thermogenesis and fatty acid oxidation in adults and leukocyte migration and tumor necrosis factor production in aged mice. Notably, Ppara and Sfpi1 emerged as potential master regulators of co-expressed genes in burn AT of adult and aged mice, respectively. Our findings highlight age-specific differences in burn-induced AT responses and uncover potential molecular regulators that may inform targeted therapeutic strategies to mitigate the post-burn stress response.