Effective Therapies for Ocular Injuries by Vesicating Agents

描述（由申请人提供）：尽管暴露于起泡剂硫芥（SM）、氮芥（NM）和砷起泡剂路易氏剂（LEW）会造成迫在眉睫的威胁和毁灭性的眼部损伤，但在大规模伤亡的情况下，有效的治疗方法仍然难以实现。这主要是由于缺乏成熟的动物损伤模型，对眼部病变的发病机制和相关机制的研究不足。这种研究的另一个实际限制是缺乏起泡剂和在实验环境中使用它们的适当设施。因此，本申请的首要目标是开发与建立发泡剂（NM、SM和LEW）诱导的眼损伤模型相关的综合策略并定义相关机制。一旦实现，这将有助于我们使用机制驱动的治疗药物进行疗效研究;这是本申请的第二个主要目标。在兔角膜器官培养中的初步研究支持了我们的两个目标，显示NM暴露增加上皮厚度、凋亡细胞死亡、上皮-基质分离、VEGF、考克斯-2和MMP-9的水平;而批准的处方药抗炎药地塞米松和抗生素强力霉素（MMP抑制剂）以及天然试剂水飞蓟宾显著降低NM诱导的上皮厚度、上皮-基质分离以及VEGF、考克斯-2和MMP-9水平，尽管是在不同的层面上。总体而言，基于上述基本原理和初步数据，本申请是可交付成果驱动的，旨在检验以下假设：发泡剂NM、SM和LEW引起眼损伤的大部分机制遵循相似的途径，通过靶向这些途径，我们将能够开发针对发泡剂诱导的眼损伤的有效和广谱疗法。我们的具体目标是：1.使用起泡剂充分表征并建立眼损伤模型。具体而言，我们将评估生物学改变并确定相关机制，包括与炎症，水疱和新血管形成相关的机制。2.评价机制靶向药物（单独或联合）和水飞蓟宾（一种对起泡剂诱导的皮肤损伤具有较强治疗效果的多效性药物）治疗眼部疾病的疗效。 伤害NM 3.评估并确定目标2中发现的有效治疗NM诱导的眼损伤的药物的有效性，以挽救SM和LEW诱导的兔眼损伤。我们预计，拟议的研究将建立有用的眼损伤模型与水疱剂，并增加我们的理解的机制/S的NM-，SM-和LEW诱导的眼损伤。此外，这些全面的研究工作将确定有效的基于广谱机制的药物，这些药物单独或组合可提供有效的治疗方法，以对抗人类中由起泡剂诱导的眼损伤。 公共卫生相关性：尽管作为战争和恐怖分子试剂的使用的威胁迫在眉睫，但针对起泡化学试剂、硫芥（SM）-、氮芥（NM）-和路易氏剂（LEW）-诱导的毁灭性眼损伤的有效疗法仍然难以捉摸。这主要是由于缺乏已建立的损伤模型，以及对眼部病变的发病机制和相关机制的研究不足;在实验环境中使用这些药物的工程设施不适当是另一个问题。因此，本申请的目标是：a）开发NM-、SM-和LEW-诱导的眼损伤模型，并定义相关的机制，和B）用机制驱动的治疗剂进行功效研究。我们的目标得到了兔角膜器官培养的初步研究的支持，这些研究表明NM-引起炎症、水疱和血管生成反应的增加，这些反应通过使用批准的抗炎和抗生素药物以及天然黄烷酮水飞蓟宾治疗而显著降低。总的来说，本申请将测试这样的假设，即由起泡剂引起的眼损伤中涉及的大多数机制遵循类似的途径，并且通过靶向这些途径，我们将能够开发针对起泡剂诱导的眼损伤的有效且广谱的疗法。我们预计，这里的研究结果将增加我们对NM、SM和LEW诱导的眼损伤的作用机制的理解，并确定基于有效机制的广谱药物，这些药物单独或联合使用，可为人类起泡剂诱导的眼损伤提供安全有效的治疗。

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.