Understanding Mustard Vesicants Distribution and Toxicity in the Eye Using In Vivo and In Silico Models

项目摘要 芥子气起泡剂硫芥子气（SM）是历史上使用最广泛的战争剂， 战争和恐怖主义的潜在代理人眼是SM及其类似物氮芥最敏感的器官 (NM)暴露，导致毁灭性的双相损伤称为芥子气角膜病变。剂量和时间- 相关的症状严重程度和这种双相损伤的机制，包括对最受影响的损伤 角膜和可能的其它眼组织的影响还没有很好的了解。此外，芥末在多大程度上 水疱剂在眼睛的各种组织中的分布、运输和持续存在在很大程度上是未知的，这可能进一步 确定组织特异性毒性作用、损伤进展和修复过程。这些研究可以帮助 确定有效的治疗策略，以对抗组织特异性急性和长期眼部并发症 芥末疱剂暴露造成的，这是难以捉摸的为了解决这一知识差距，我们将使用一个协同 体内-计算机模拟方法，以确定芥子气起泡剂的眼部分布和随后的角膜损伤， 修复机制我们将利用CFD研究公司现有的电子兔眼模型， 用于表征各种化学威胁剂的分布相关毒性。根据公布的数据， 初步的计算机模拟研究显示NM穿透远超过角膜。我们假设 协同新颖体内-计算机模拟方法将有助于理解芥子类水疱剂如何在健康 眼睛和眼损伤的发展，持续，恢复或复发的机制，作为长期 并发症为了支持这一假设，提出的具体目标将首先计算模型NM（SM 实验室环境中使用的替代物）相关的眼损伤，然后将其转化为模拟SM暴露。目的 1：确定各种暴露情况下氮芥的眼部分布。计算 模型将根据家兔体内研究的NM毒代动力学（TK）曲线进行验证，然后外推至 实验测试范围，以获得非测试NM暴露场景的TK特征。目标2：确定 NM诱导的角膜损伤进展和修复的相关生物标志物、机制和途径。 时间和剂量依赖性转录组学和生物标志物评价以及NM TK的组合 根据目标1提出的分析，可能有助于确定早期和 与水疱性双相角膜病变相关的晚期角膜损伤发展、进展和修复。目标3： 翻译计算机模拟NM兔眼模型以模拟SM暴露。初步评估 可以评估Silico SM角膜损伤模型与NM的比较，并作为次要的 变更或设计动物研究，以便在未来研究中进行额外验证。结合计算机模拟 体内研究可以作为一种更有效和更有洞察力的新方法来评估 芥子类水疱剂眼部病理学的组织特异性毒性机制，并确定 治疗策略，可以改善临床芥末角膜病变症状，促进伤口愈合。

Project Summary Mustard vesicating agent sulfur mustard (SM) is the most extensively used warfare agent in history and remains a potential agent of warfare and terrorism. Eye is the most sensitive organ to SM and its analog nitrogen mustard (NM) exposure, resulting in devastating biphasic injury called Mustard Gas Keratopathy. The dose- and time- related severity of symptoms and the mechanism of this biphasic injury involving damage to the most affected cornea and possibly to other ocular tissues is not well understood. In addition, the extent to which mustard vesicants distribute, transport, and persist in various tissues of the eye is largely unknown, which could further identify tissue specific toxicity effects, injury progression and repair process. Such studies could aid in the identification of effective treatment strategies to counter tissue specific acute and long-term ocular complications from mustard vesicants’ exposure, which are elusive. To address this knowledge gap, we will use a synergistic in vivo-in silico approach to determine mustard vesicant ocular distribution and the ensuing corneal injury and repair mechanisms. We will leverage CFD Research’s existing in silico rabbit ocular model that is currently being used to characterize the distribution related toxicity of various chemical threat agents. Using the published data, preliminary in silico modeling studies show that NM penetrates well beyond the cornea. We hypothesize that a synergistic novel in vivo-in silico approach will assist in understanding how mustard vesicants distribute in healthy eyes and the mechanism by which the ocular injury develops, persists, and recovers or reappears as long-term complications. To support this hypothesis, the proposed specific aims will first computationally model NM (SM surrogate used in laboratory settings) related ocular injury and then translate this to simulate SM exposure. Aim 1: Determine Ocular Distribution of Nitrogen Mustard for Various Exposure Scenarios. The computational model will be validated on NM toxicokinetic (TK) profiles from in vivo rabbit studies and then extrapolated within the experimentally tested range to obtain TK profiles for non-tested NM exposure scenarios. Aim 2: Identify Relevant Biomarkers, Mechanisms and Pathways of NM-Induced Corneal Injury Progression and Repair. The combination of time- and dose-dependent transcriptomics and biomarker evaluation, as well as the NM TK analysis proposed under Aim 1, may contribute to determine the pathological mechanisms underlying early and late corneal injury development, progression, and repair related to vesicant bi-phasic keratopathy. Aim 3: Translation of the In Silico NM Rabbit Ocular Model to Simulate SM Exposure. The preliminary evaluations could assess comparisons of in Silico SM corneal injury model with NM, and serve as a vital step for minor alterations or in designing animal studies for additional validation under future studies. A combination of in silico and in vivo studies could serve as a more efficient and potentially more insightful novel approach to evaluate tissue-specific toxicity mechanisms underlying the mustard vesicants’ ocular pathology, and to identify therapeutic strategies that can improve the clinical mustard keratopathy symptoms and enhance wound healing.