Therapeutic Intervention of Lewisite-Mediated Cutaneous Blistering-Inflammation

路易氏剂介导的皮肤起泡炎症的治疗干预

• 批准号: 8927140
• 负责人: Mohammad Athar
• 金额: $ 7.35万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-18 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8927140
• 关键词: Acute; Amines; Animal Model; Animals; Antidotes; Arsenic; Arsenicals; Attenuated; Biological; Biological Assay; Biological Markers; Biopsy; Blood capillaries; British; Bulla; Carrier Proteins; Cathepsins; Cell Degranulation; Cells; Chemical Agents; Chemical Warfare; Chemicals; Clinical Trials; Cutaneous; DNA Alkylation; DNA Damage; Data; Dermal; Dermatology; Development; Diet; Digestion; Dinoprostone; Edema; Effectiveness; Eicosanoids; Epidermis; Erythema; Event; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Exposure to; Extravasation; Filament; Focal Adhesion Kinase 1; Future; Glutathione; Glycerol; Hair follicle structure; Health; Health protection; Human; Hyperplasia; In Vitro; Inbred HRS Mice; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Intercept; Interferons; Interleukin-6; Kinetics; Lead; Leukocytes; Liquid substance; Mediating; Modeling; Molecular; Molecular Biology; Molecular Target; Mus; Mustard Agent; Outcome; Oxidative Stress; Pain; Pathogenesis; Penetration; Peptide Hydrolases; Phosphotransferases; Poison; Population; Process; Production; Protective Agents; Proteins; Reaction; Reactive Nitrogen Species; Reactive Oxygen Species; Regulation; Regulatory Pathway; Relative (related person); Research; Research Proposals; Rupture; Series; Signal Pathway; Signal Transduction; Skin; Subcutaneous Tissue; System; Testing; Therapeutic; Therapeutic Intervention; Thick; Tight Junctions; Toxic effect; Toxicology; Translational Research; United States National Institutes of Health; Vesicants; War; Water; World War I; base; capillary; chemical threat; chemokine; cytokine; in vivo; interest; keratinocyte; lewisite; macrophage; mast cell; novel; occludin; prevent; prospective; response; skin lesion; small molecule; ubiquitin ligase; water channel

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.

描述（由申请人提供）：路易氏剂已被确定为一种可用于化学战的潜在威胁化学品。局部接触路易斯会导致皮肤起泡和炎症，可能会很严重且疼痛。在此应用中，我们将测试以下假设：刘易斯通过穿透皮肤，破坏与紧密连接和水/甘油运输相关的蛋白质，从而破坏皮肤屏障功能。这些作用是通过激活 hippo 信号通路、FAK 激酶、泛素连接酶、组织蛋白酶等蛋白酶和其他溶菌酶来介导的。急性炎症是通过激活未折叠蛋白反应（UPR）信号介导的，而该信号由砷依赖性活性氧（ROS）的产生和DNA损伤反应信号的激活所触发。这些复杂的信号通路之间的串扰导致了疼痛性水疱和炎症的发病机制。用小分子阻断这些分子靶标可能会阻止这些效应。目标 1 将通过研究刘易斯对表皮中紧密连接蛋白（如紧密连接蛋白、闭合蛋白、闭合小带 (ZO) 等）的影响，研究刘易斯对皮肤屏障功能破坏和起泡的影响。此外，还将研究参与水/甘油运输调节的蛋白质，即水通道蛋白。还将通过确定炎症反应的动力学并在其峰值时评估增生和炎症细胞浸润来研究路易斯对急性皮肤炎症反应的影响。然后，我们将使用路易斯处理的Ptch1+/-/SKH-1无毛小鼠的皮肤活检来测定促炎介质、IL-6、干扰素-β、前列腺素E2、ROS、RNS（包括NO）等。我们将确定路易斯调节的 UPR 信号是否调节这些促炎作用。在目标 2 中，我们将筛选饮食和合成测试剂对描述屏障功能、水泡和炎症的生物标志物的有效性。在此目的中，将根据已知的阻断涉及屏障功能/起泡/炎症的分子靶点的潜力，从一系列膳食和合成化学品中选择两种最有效的药物。确定治疗窗后，将评估这些选定药物的作用动力学、逆转路易斯诱导的分子变化的能力以及潜在的皮肤炎症/水疱。因此，该提案将采用一种新型小鼠模型来揭示路易斯的分子机制，以开发基于机制的解毒剂/疗法。通过提交前瞻性申请（R01 或 U01），可以轻松地将小分子候选先导化合物用于进一步的小鼠研究，并最终根据其已知的毒性特征和用于其他条件的情况进行临床试验。如果大量人口接触战争威胁化学品，拟议研究的结果可能会对人类健康保护产生重大影响。