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.

项目概要 硫芥 (SM) 仍然对平民和军人构成重大威胁。皮肤接触 SM 会导致 红斑，随后受影响区域出现水肿和大水疱，愈合时间较长。顺序 这种损伤中细胞死亡和脱离的方式仍未解决，部分原因是体外和 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和 根据天然皮肤的生理学，非常需要开发具有多种细胞类型的生物工程皮肤， 适当的生理潜力。评估生物打印皮肤作为离体类器官（器官组织 等）已证明其在体外 2 个月以上仍能保持其层状结构。评估 通过 RNA 和蛋白质分析，皮肤类器官显示出与皮肤类器官相似的生理反应 正常人类皮肤。这些令人鼓舞的结果表明，这种皮肤结构正在重现人体器官 我们建议使用通过本补充提供的资金进一步扩大生物打印人体皮肤 建立化学烧伤模型。我们将使用生物打印皮肤来模拟接触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。 根据我们迄今为止生成的数据，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效应模型。总体目标 该补充提案的目的是生成主要数据证明，表明生物打印皮肤会做出反应 以与 SM 的已知效果一致的方式来应对化学损伤。我们的研究将验证剂量 和暴露时间，用于模拟人体对 SM 的体外反应，包括组织学、qRT-PCR、 和 ELIZA 分析。这些研究将重点证明生物打印皮肤的行为方式是 与特定目标 1 一致，将描述硫芥暴露的病理后果。 具体目标 2 将寻找受 SM 诱导损伤影响的细胞途径。这些研究将验证 使用生物打印皮肤作为 SM 毒性的可行离体模型，这将实现高精度分析， 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，它表明生物打印的皮肤可以发展成人类皮肤的一般替代品 皮肤病理生理学可用于研究其他类型的损伤。最终，我们相信该系统将识别 暴露和治疗效果的生物标志物，以及作为发现和治疗效果的有用方式 优化一系列化学和生物武器的医疗对策。