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

项目总结 硫磺芥子(SM)仍然对平民和军人构成重大威胁。皮肤暴露于SM诱导 红斑，紧随其后的是受影响区域的水肿和大水泡，愈合时间延长。该序列 而这种损伤中细胞死亡和脱落的方式仍未解决，部分原因是体外培养不完全和 临床前模型。由于体外3D皮肤模型的目标是复制真实的解剖结构和 天然皮肤的生理学，迫切需要开发具有多种细胞类型和 适当的生理潜力。生物打印皮肤作为体外有机物(器官组织)的评估 等价物)已经证明它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，皮肤有机化合物显示出类似于在 正常的人类皮肤。这些令人鼓舞的结果表明，这种皮肤结构是对人体器官的概括。 我们建议使用通过这一补充提供的资金来进一步扩大生物打印的人类皮肤 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接触这种已知化学物质的情况 发泡剂硫磺芥末(SM)，最终目的是显示生物打印皮肤在阐明 生物化学和病理生理途径，以努力发现生物标记物和医学对策。 基于我们到目前为止产生的数据，我们的中心假设是体外生物打印的皮肤将 化学发泡剂硫磺芥末暴露的临床病理效应模型。总目标 这一补充建议的目的是产生主要数据的证据，表明生物打印的皮肤将产生反应 对化学物质的侮辱，其方式与SM的已知影响一致。我们的研究将验证剂量 和暴露时间，以模拟人体对SM的体外反应，包括组织学、qRT-PCR、 和伊莱扎的分析。这些研究将集中于证明生物打印的皮肤的行为方式是 与特定目标1一致，将描述硫芥子气暴露的病理后果。 特定目标2将寻求确定SM诱导的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毒性的可行的体外模型，这将使高精度分析成为可能， 包括实时监测和组学研究硫芥子皮的序列和机制 受伤。此外，还表明生物印花皮肤可以发展成为人类的通用替代品。 皮肤病理生理学对研究其他类型的侮辱很有用。最终，我们相信这个系统将识别 暴露和治疗效果的生物标志物，以及作为一种有用的发现和 优化针对一系列化学和生物武器的医学对策。

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.