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: 8544981
• 负责人: Mohammad Athar
• 金额: $ 36.63万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8544981
• 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 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; chemokine; cytokine; in vivo; interest; keratinocyte; lewisite; macrophage; mast cell; novel; occludin; prevent; prospective; public health relevance; 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）：Lewisite已被确定为可以在化学战中使用的潜在威胁化学物质。局部暴露于路易斯特岩会导致皮肤起泡和炎症，这可能严重而痛苦。在此应用中，我们将检验以下假设：刘易斯铁矿通过穿透皮肤，破坏皮肤屏障的功能，这是由于破坏与紧密连接和水/甘油传输相关的蛋白质的破坏。这些作用是通过河马信号通路，FAK激酶，泛素连接酶，蛋白酶（例如组织蛋白酶和其他溶菌酶蛋白酶）的激活来介导的。急性炎症是通过激活未折叠的蛋白质反应（UPR）信号传导来介导的，该信号传导是由砷依赖性的活性氧（ROS）产生和DNA损伤响应信号传导激活的激活而触发的。这些错综复杂的信号通路之间的串扰导致疼痛水泡和炎症的发病机理。通过小分子阻止这些分子靶标可能会截断这些作用。 AIM 1将通过研究表皮中的紧密连接蛋白（例如Claudins，occludin，Zonula occludens（ZO）等）来研究Lewisite对皮肤屏障功能的破坏和起泡的影响。此外，将研究参与水/甘油转运的调节的蛋白质，即水通道蛋白。路易斯特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，并在其峰值上评估增生和炎症细胞浸润。然后，我们将使用Lewisite处理过的PTCH1+/ - /SKH-1无毛小鼠的皮肤活检，分析促炎性介体IL-6，Interferon-？，Prostaglandin E2，ROS，RN（包括NO）等。我们将确定Lewisite调节的UPR信号是否调节这些促炎作用。在AIM 2中，我们将筛选饮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。从一系列饮食和合成化学物质中选择的两种最有效的药物基于其已知的潜在阻断参与屏障功能/泡沫/炎症的分子靶标的潜力，将进一步评估此目标。在定义了治疗窗口后，将评估这些精选药物的分子变化和潜在的皮肤炎症/起泡的能力，将评估这些精选药物的潜在炎症/起泡的能力。因此，该建议将采用一种新型的鼠模型来揭示路易斯特岩的分子机制，以开发基于机制的解毒剂/治疗。小分子候选铅化合物可以通过提交前瞻性应用（R01或U01），最终根据其已知的毒性概况和其他条件使用，轻松地进行进一步的鼠研究。在大规模人口暴露于战争威胁化学物质的情况下，拟议研究的结果可能会对人类健康保护产生重大影响。