Effective Therapies for Ocular Injuries by Vesicating Agents

描述(由申请人提供)：尽管接触发泡剂硫芥酸(SM)、氮芥酸(NM)和砷发泡剂路易威特(LOW)会造成迫在眉睫的威胁和毁灭性的眼部损伤，但在发生大规模伤亡的情况下，有效的治疗方法仍然难以找到。这主要是由于缺乏已建立的动物损伤模型，对眼部损伤的发病机制和相关机制的研究也不充分/S。此类研究的另一个实际限制也是缺乏可用的发泡剂和适当的设施在实验环境中使用它们。因此，这项应用的首要目标是开发一种与建立发泡剂(NM、SM和LEW)诱导的眼损伤模型相关的综合策略，并确定相关的机制。一旦实现，这将有助于我们进行机制驱动的治疗剂的疗效研究；这是这项应用的第二个主要目标。我们的两个目标都得到了兔角膜器官培养的初步研究的支持，研究表明，NM暴露增加了角膜上皮厚度、细胞凋亡、上皮-间质分离、血管内皮生长因子、环氧合酶-2和基质金属蛋白酶-9的水平；批准的处方药地塞米松和抗生素多西环素(一种基质金属蛋白酶抑制剂)以及天然药物水飞蓟宾显著减少了近视眼诱导的上皮厚度、上皮-间质分离以及血管内皮生长因子、环氧合酶-2和基质金属蛋白酶-9的水平，尽管水平不同。总体而言，基于上述基本原理和初步数据，这项应用是可交付成果驱动的，并建议测试这样一种假设，即发泡剂NM、SM和LEW引起的眼损伤涉及的大多数机制遵循相似的途径，并且通过针对这些途径，我们将能够开发针对发泡剂引起的眼损伤的有效和广谱治疗。我们的具体目标是：1.充分表征和建立具有起泡剂的眼损伤模型。具体地说，我们将评估生物变化并确定相关机制，包括与炎症、水泡和新生血管有关的机制。2.评价机制靶向药物(单独或联合使用)和水飞蓟宾(一种对起泡剂引起的皮肤损伤有很强疗效的多效剂)治疗眼部的疗效 NM造成的伤害。3.评价和确定在Aim 2中发现的治疗NM诱导的眼损伤的有效药物/S对SM和LEW诱导的眼损伤的抢救效果。我们期望所提出的研究将建立有用的发泡剂眼损伤模型，并加深我们对NM-、SM-和LEW诱导的眼损伤作用机制的理解。此外，这些全面的研究工作将确定有效的广谱机制制剂/S，单独或联合提供有效的治疗方法来对抗发泡剂引起的人类眼损伤。 与公共卫生相关：尽管被用作战争和恐怖分子毒剂的威胁迫在眉睫，但针对硫芥子气(SM)、氮芥子气(NM)和路易氏剂(LOW)引起的毁灭性眼损伤的有效治疗方法仍然难以找到。这主要是由于缺乏已建立的损伤模型，以及对眼部损伤的发病机制和相关机制的研究不足/S；不适当的工程设施在实验环境中使用这些制剂是另一个令人担忧的问题。因此，这项应用的目标是：a)建立NM、SM和LEW诱导的眼损伤模型，并定义相关的机制，以及b)使用机制驱动的治疗剂进行疗效研究。我们的目标得到了兔角膜器官培养的初步研究的支持，研究表明，NM-导致炎症、水飞蓟素和血管生成反应的增加，而批准的抗炎和抗生素药物以及天然黄烷酮水飞蓟宾的治疗显著降低了这些反应。总体而言，本申请将检验以下假设，即发泡剂引起的眼部损伤的大多数机制遵循相似的途径，通过针对这些途径，我们将能够开发有效和广谱的治疗方法来对抗发泡剂引起的眼部损伤。我们期望本文的研究结果将加深我们对NM、SM和LEW诱导的眼损伤作用机制/S的理解，并确定有效的基于机制的广谱制剂/S，单独或联合使用，提供安全有效的治疗方法来对抗发泡剂诱导的人类眼损伤。

DESCRIPTION (provided by applicant): Despite the imminent threat and devastating ocular injuries by exposure to vesicating agents sulfur mustard (SM), nitrogen mustard (NM) and arsenical vesicant, lewisite (LEW), effective therapies in case of a mass casualty remain elusive. This is mainly due to the lack of established animal injury models and inadequate study on the pathogenesis of eye lesions and associated mechanism/s. An additional practical limitation for such studies has also been a lack of availability of vesicating agents and appropriate facilities to use them in experimental settings. Accordingly, the first and foremost goal of this application is to develop a comprehensive strategy related to establishing vesicant agents (NM, SM and LEW)-induced ocular injury models and define the associated mechanisms. Once achieved, this will help us perform efficacy studies with mechanism-driven therapeutic agents; this is the second major goal of this application. Both our goals are supported by preliminary studies in rabbit corneal organ culture showing that NM exposure increases epithelial thickness, apoptotic cell death, epithelial-stromal separation, levels of VEGF, COX-2 and MMP-9; and that approved prescription agents anti-inflammatory drug dexamethasone and antibiotic doxycycline (an MMP inhibitor) as well as a natural agent silibinin significantly reduce NM-induced epithelial thickness, epithelial-stromal separation, and VEGF, COX-2 and MMP-9 levels, albeit at different levels. Overall, based on above rationale and preliminary data, this application is deliverables driven and proposes to test the hypothesis that most of the mechanisms involved in ocular injuries by vesicating agents NM, SM and LEW follow similar pathways, and that , by targeting those pathways, we would be able to develop effective and broad-spectrum therapies against vesicants-induced ocular injuries. Our specific aims are: 1. Fully characterize and establish ocular injury models with vesicating agents. Specifically, we will assess biological alterations and identify associated mechanisms including those related to inflammation, vesication and neovascularization. 2. Evaluate the efficacy of mechanism-targeted agents (alone or in combination) and silibinin (a pleiotropic agent with strong therapeutic efficacy against blistering agents-induced skin injuries) in treating the ocular injuries by NM. 3. Assess and establish the efficacy of the agent/s found effective in treating NM-induced ocular injuries in Aim 2, for the rescue of SM- and LEW-induced ocular injuries in rabbit. We anticipate that the proposed studies will establish useful ocular injury models with vesicants, and add to our understanding of the mechanism/s of action of NM-, SM- and LEW-induced ocular injuries. In addition, these comprehensive study efforts will identify effective broad spectrum mechanism-based agent/s that alone or in combination provide effective therapies against vesicating agents-induced ocular injuries in humans. PUBLIC HEALTH RELEVANCE: Despite the imminent threat of use as warfare and terrorist agents, effective therapies against vesicating chemical agents, sulfur mustard (SM)-, nitrogen mustard (NM)- and lewisite (LEW)-induced devastating ocular injuries, remain elusive. This is mainly due to the lack of established injury models, and inadequate study on the pathogenesis of eye lesions and associated mechanism/s; inappropriate engineering facilities to use these agents in experimental settings are an additional concern. Accordingly, the goals of this application are: a) develop NM-, SM- and LEW-induced ocular injury models, and define the associated mechanisms, and b) perform efficacy studies with mechanism-driven therapeutic agents. Our goals are supported by preliminary studies in rabbit corneal organ culture showing that NM-causes an increase in the inflammatory, vesication, and angiogenic responses, which are significantly reduced by the treatment with approved anti-inflammatory and antibiotic drugs as well as a natural flavanone, silibinin. Overall, the present application will test the hypothesi that most of the mechanisms involved in ocular injuries by vesicating agents follow similar pathways, and that, by targeting those pathways, we would be able to develop effective and broad-spectrum therapies against vesicants-induced ocular injuries. We anticipate that the study outcomes here will add to our understanding of the mechanism/s of action of NM-, SM- and LEW-induced ocular injuries, and identify effective mechanism-based broad-spectrum agent/s that, alone or in combination, provide safe and effective therapies against vesicating agents-induced ocular injuries in humans.