Therapeutic Intervention of Lewisite-Mediated Cutaneous Blistering-Inflammation

描述（由申请人提供）：路易斯特已被确定为一种潜在的威胁化学品，可用于化学战。局部暴露于刘易斯导致皮肤起泡和炎症，这可能是严重和痛苦的。在这个应用中，我们将测试路易斯体通过穿透皮肤，破坏皮肤屏障功能的假设，这是与紧密连接和水/甘油运输相关的蛋白质被破坏的结果。这些作用是通过激活hippo信号通路、FAK激酶、泛素连接酶、蛋白酶（如Cathepsin）和其他溶菌酶来介导的。急性炎症是通过砷依赖性活性氧（ROS）产生引发的未折叠蛋白反应（UPR）信号的激活和DNA损伤反应信号的激活介导的。这些错综复杂的信号通路之间的相互作用导致了疼痛性水泡和炎症的发病机制。用小分子阻断这些分子靶点可能会阻断这些作用。目的1将通过研究路易斯体对表皮紧密连接蛋白（如claudin、occludin、zonula occludens （ZO）等）的影响，研究路易斯体对皮肤屏障功能破坏和起泡的影响。此外，还将研究参与调节水/甘油运输的蛋白质，即水通道蛋白。路易斯体对急性皮肤炎症反应的影响也将通过确定炎症反应的动力学来研究，并在其高峰期评估增生和炎症细胞浸润。然后，我们将检测促炎介质，IL-6，干扰素-？，前列腺素E2， ROS， RNS（包括NO）等，使用lewisite处理的Ptch1+/-/SKH-1无毛小鼠的皮肤活检。我们将确定路易斯体调节的UPR信号是否调节这些促炎作用。在目标2中，我们将筛选膳食和合成测试剂对描述屏障功能、起泡和炎症的生物标志物的有效性。根据已知阻断屏障功能/起泡/炎症的分子靶点的潜力，从一系列膳食和合成化学物质中选择两种最有效的药物将在此目的中进一步评估。在确定治疗窗口期后，将评估这些选定药物的作用动力学、逆转lewis诱导的分子变化和潜在皮肤炎症/起泡的能力。因此，本研究将采用一种新的小鼠模型来揭示路易斯体的分子机制，从而开发基于机制的解毒剂/治疗方法。小分子候选铅化合物可以很容易地通过提交前瞻性申请（R01或U01）进行进一步的小鼠研究，最后根据其已知的毒性特征和用于其他条件进行临床试验。拟议的研究结果很可能在大量人口接触战争威胁化学品的情况下对人类健康的保护产生重大影响。

DESCRIPTION (provided by applicant): Lewisite has been identified as a potential threat chemical which could be used in chemical warfare. Topical exposure to lewisite results in cutaneous blistering and inflammation, which may be severe and painful. In this application, we will test the hypothesis that lewisite, by penetrating skin, ruptures the cutaneous barrier functions as a consequence of disruption of proteins associated with tight junctions and water/glycerin transport. These effects are mediated via activation of hippo signaling pathway, FAK kinases, ubiquitin ligases, proteases such as Cathepsin, and other lysozymal proteases. The acute inflammation is mediated through the activation of unfolded protein response (UPR) signaling triggered by arsenic-dependent reactive oxygen species (ROS) production and activation of DNA damage response signaling. Crosstalk between these intricate signaling pathways results in the pathogenesis of painful blisters and inflammation. Blocking these molecular targets by small molecules may intercept these effects. Aim 1 will investigate the effects of lewisite on disruption of skin barrier function and blistering by studying its effects o tight junction proteins such as claudins, occludin, zonula occludens (ZO), etc. in epidermis. In addition, proteins involved in the regulation of water/glycerin transport, the aquaporins, will be studied. The effects of lewisite on acute cutaneous inflammatory response will also be investigated by determining the kinetics of inflammatory responses and, at their peak, assessing hyperplasia and inflammatory cell infiltration. Then, we will assay the pro-inflammatory mediators, IL-6, interferon-?, prostaglandin E2, ROS, RNS (including NO) etc. using skin biopsies from lewisite- treated Ptch1+/-/SKH-1 hairless mice. We will determine whether lewisite-modulated UPR signaling regulates these pro-inflammatory effects. In Aim 2, we will screen the effectiveness of dietary and synthetic test agents on biomarkers depicting barrier function, blistering, and inflammation. The two most efficacious agents chosen from a series of dietary and synthetic chemicals based on their known potential to block molecular targets involved in barrier function/blistering/inflammation will be evaluated further in this aim. After defining the therapeutic window, kinetics of action, ability to reverse lewisite-induced molecular changes and underlying cutaneous inflammation/blistering by these select agents will be assessed. Thus, this proposal will employ a novel murine model to unravel the molecular mechanism of lewisite in order to develop mechanism-based antidotes/therapy. Small molecule candidate lead-compounds can easily be taken to further murine studies by submitting a prospective application (R01 or U01) and finally to clinical trials based on their known toxicity profile and use for other conditions. The outcome of the proposed research is likely to have a significant impact on human health protection in the event of mass population exposure to war threat chemicals.