The Initiation of Vesicant Skin Injury at a Single Cell Level

摘要 硫磺芥末(芥子气)和路易斯安特等军火库是化学战或恐怖分子的主要威胁。 探员们。尽管化学结构不同，但这两种药物都会导致相似的皮肤损伤，出现大水泡。 以及愈合缓慢的伤口，引发了共同致病机制的幽灵。在这项提案中，我们将 研究氧化性皮肤损伤在芥末和砷发泡剂损伤中的共同机制， 分别使用氮芥末和苯基氧化砷作为硫磺芥末和路易石的替代品。 虽然发泡剂的研究已经取得了重大进展，但我们仍然没有有效的对抗剂。 这可能部分是由于对发泡剂的发病机制缺乏充分的了解。最重要的假设 这一提议的核心是，在亚细胞和单细胞水平上定义发泡剂在体内的早期影响将得到完善 我们对氧化应激在发泡性皮肤损伤中的作用的理解，并导致新的治疗策略。 我们还将检验这样一种假设，即可比酰胺，一种高效和多功能的抗氧化剂，中和两者 活性氧和氮物种，将是有效的反作用剂和更好地了解 氧化应激在氮芥末和氧化苯砷诱导的发泡剂损伤中的作用。具体的 目的是：1)了解氮芥末和氧化苯砷致皮肤损伤的始动机制。 在单个细胞水平上。我们将使用体内荧光寿命成像(FLIM)和单细胞RNA测序 以期对发泡损伤的发生和氧化应激在发病机制中的作用有一个新的认识。2)至 确定可比酰胺对氮芥末和苯砷氧化物诱导的发泡剂的疗效 皮肤受伤。我们将测试可比酰胺对氮芥末和苯基砷氧化物诱导的效果。 发泡剂对小鼠皮肤的损伤作用。由于老鼠和人类皮肤的不同，我们还将研究发泡剂 移植的人类皮肤的机制。这些研究具有重要意义和创新性，因为它们使用了 ART工具来研究我们对氧化应激在芥末起始过程中的作用的认识上的差距 和砷发泡剂损伤，因为他们第一次测试了椰子酰胺，一种高效抗氧化剂 这对于活性氧和氮物种来说是双官能团。

Abstract Sulfur mustard (mustard gas) and arsenicals such as lewisite are major threats as chemical warfare or terrorist agents. Despite different chemical structures, both types of agents lead to similar skin damage with large blisters and wounds that heal slowly, raising the specter of shared pathogenic mechanisms. In this proposal, we will investigate the role of oxidant skin injury as a shared mechanism in mustard and arsenical vesicant injury, employing nitrogen mustard and phenylarsine oxide as surrogates for sulfur mustard and lewisite, respectively. Although major advances have been made in vesicant research, we still do not have effective counter agents. This may be in part due to lack of full understanding of the pathogenesis of vesicants. The overarching hypothesis of this proposal is that defining the early effects of vesicants in vivo at subcellular and single cell levels will refine our understanding of the role of oxidative stress in vesicant skin injury and lead to new therapeutic strategies. We will also test the hypothesis that cobinamide, a highly effective and versatile antioxidant that neutralizes both reactive oxygen and nitrogen species, will be effective as a counter-agent and as means to understand better the role of oxidative stress in nitrogen mustard- and phenylarsine oxide-induced vesicant injury. The specific aims are: 1) To understand the initiation of nitrogen mustard- and phenylarsine oxide-induced skin injury at a single cell level. We will use in vivo fluorescence lifetime imaging (FLIM) and single cell RNA sequencing to gain a new view on the initiation of vesicant injury and the role of oxidative stress in the pathogenesis. 2) To define the efficacy of cobinamide against nitrogen mustard- and phenylarsine oxide-induced vesicant skin injury. We will test the efficacy of cobinamide against nitrogen mustard- and phenylarsine oxide-induced vesicant skin injury in mice. Due to differences between mouse and human skin, we will also investigate vesicant mechanisms in transplanted human skin. These studies are significant and innovative because they use state of the art tools to investigate a gap in our knowledge about the role of oxidative stress in the initiation of mustard and arsenical vesicant injury and because they test for the first time cobinamide, a highly effective antioxidant that is bifunctional for reactive oxygen and nitrogen species.