Characterization of an Ex Vivo Bioprinted Skin Model of Sulfur Mustard Injury

硫芥损伤离体生物打印皮肤模型的表征

• 批准号: 10228424
• 负责人: ADAM JORGENSEN
• 金额: $ 5万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-17 至 2021-11-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10228424
• 关键词: 3-Dimensional; Address; Adherence; Adipocytes; Affect; Anatomy; Animal Model; Animals; Area; Basement membrane; Biochemical; Biological; Biological Markers; Biomedical Engineering; Biomedical Research; Biomimetics; Bulla; Burn injury; CD14 gene; Cell Culture Techniques; Cell Death; Cells; Cessation of life; Chemical Models; Chemical Warfare Agents; Chemical Weapons; Chemicals; Cicatrix; Clinical; Collagen Type VII; Complex; Data; Dermal; Dermatologic; Development; Disease; Dose; Edema; Elements; Endothelial Cells; Epidermal Ridges; Epidermis; Erythema; Exposure to; Fibroblasts; Functional disorder; Funding; Gene Expression; Genetic Transcription; Goals; Histologic; Human; In Vitro; Inflammation; Inflammation Mediators; Injury; Investigation; Metalloproteases; Military Personnel; Modality; Modeling; Mustard Gas; Organ; Organoids; Pathologic; Pathway interactions; Physiological; Physiology; Pigmentation physiologic function; Poly(ADP-ribose) Polymerases; Population; Pre-Clinical Model; Protein Analysis; Quantitative Reverse Transcriptase PCR; RNA analysis; Research Institute; Scanning Electron Microscopy; Signaling Molecule; Skin; Skin injury; Structural Protein; Structure; System; Techniques; Testing; Time; Tissue Engineering; Tissues; Toxic effect; Treatment Efficacy; Validation; Vascularization; Vesicants; World War I; base; bioprinting; bioweapon; cell type; cytokine; cytotoxicity; extracellular; healing; improved; in vitro Model; keratinocyte; laminin-5; medical countermeasure; melanocyte; monocyte; novel; organizational structure; pre-clinical; protein expression; prototype; real time monitoring; research study; response; skin barrier; three dimensional structure; two-dimensional

PROJECT SUMMARY Sulfur Mustard (SM) remains a significant threat to civilian and military populations. Skin exposure to SM induces erythema, followed by edema and large blisters in the affected area, with prolonged healing times. The sequence and manner of cell death and detachment in this injury are still unresolved, in part due to incomplete in vitro and preclinical models. Since the goal of an ex vivo 3D skin model is to replicate the authentic anatomy and physiology of native skin, there is an immense need to develop bioengineered skin with multiple cell types and appropriate physiological potential. Assessment of bioprinted skin as an ex vivo organoid (organ tissue equivalent) has demonstrated that it maintains its layered structure for over 2 months in vitro. Assessment of skin organoids by RNA and protein analysis demonstrate a physiological response similar to those seen in normal human skin. These encouraging results suggest that this skin construct is recapitulating the human organ and we propose to use the funds available through this supplement to further expand the bioprinted human skin into a model of chemical burn injuries. We will use bioprinted skin to model exposure to the known chemical vesicant sulfur mustard (SM), with the ultimate goal of showing the full utility of the bioprinted skin in elucidating biochemical and pathophysiological pathways in an effort to discover biomarkers and medical countermeasures. Based on the data we have generated to date, our central hypothesis is that ex vivo bioprinted skin will model clinical pathological effects of exposure to the chemical vesicant sulfur mustard. The overall goal of this supplemental proposal is to generate proof of principal data showing that the bioprinted skin will respond to the chemical insult in a way that is consistent with the known effects of SM. Our study will validate the dose and time of exposure for modeling the human response to SM in vitro, and will include histological, qRT-PCR, and ELIZA analysis. These studies will focus on demonstrating that the bioprinted skin behaves in a way that is consistent with Specific Aim 1 will characterize the pathological consequences of sulfur mustard exposure. Specific Aim 2 will look to identify cellular pathways affected by SM induced injury. These studies will validate the use of the bioprinted skin as a viable ex vivo model of SM toxicity which will enable high precision analysis, including real-time monitoring and -omics to elucidate the sequence and mechanism of sulfur mustard skin injuries. Furthermore, it suggests that the bioprinted skin can be developed into a general surrogate of human skin pathophysiology useful in studying other types of insults. Ultimately, we believe that this system will identify biomarkers of exposure and therapeutic efficacy, as well as serving as a useful modality to discovery and optimize medical countermeasures for a range of chemical and biological weapons.

项目摘要 硫芥（SM）仍然是对平民和军事人口的重大威胁。皮肤暴露于SM诱导 红斑，随后是受影响区域的水肿和大水泡，愈合时间延长。序列 以及这种损伤中细胞死亡和脱离的方式仍然没有解决，部分原因是体外研究不完整， 临床前模型。由于离体3D皮肤模型的目标是复制真实的解剖结构， 由于天然皮肤的生理学特性，存在开发具有多种细胞类型的生物工程皮肤的巨大需求， 适当的生理潜力。生物打印皮肤作为离体类器官（器官组织）的评估 等同物）已经证明其在体外保持其层状结构超过2个月。评估 通过RNA和蛋白质分析的皮肤类器官显示出与在 正常人的皮肤这些令人鼓舞的结果表明，这种皮肤结构是重演人体器官 我们建议利用这笔资金来进一步扩大生物打印人类皮肤， 变成了化学烧伤的模型我们将使用生物打印的皮肤来模拟暴露于已知化学物质的情况 起泡剂硫芥（SM），最终目标是展示生物打印皮肤在阐明 生物化学和病理生理学途径，努力发现生物标志物和医学对策。 根据我们迄今为止产生的数据，我们的中心假设是，离体生物打印皮肤将 暴露于化学发泡剂硫芥的临床病理学效应模型。总目标 该补充提案的一个重要目的是生成主要数据的证据，表明生物打印的皮肤将响应 以与SM的已知效果一致的方式对化学损伤进行处理。我们的研究将验证 和暴露时间，用于体外模拟人体对SM的反应，并将包括组织学，qRT-PCR， ELIZA分析这些研究将集中在证明生物打印皮肤的行为方式， 与具体目标1一致，将描述硫芥暴露的病理后果。 具体目标2将着眼于确定受SM诱导损伤影响的细胞通路。这些研究将验证 使用生物打印的皮肤作为SM毒性的可行离体模型，这将使得能够进行高精度分析， 包括实时监测和组学，以阐明硫芥皮肤的序列和机制， 受伤此外，这表明生物打印皮肤可以发展成为人类的一般替代品。 皮肤病理生理学，可用于研究其他类型的侮辱。最终，我们相信这个系统将识别 暴露和治疗功效的生物标志物，以及作为一种有用的模式来发现和 优化针对一系列化学和生物武器的医疗对策。