Oxidized lipids and microvesicle particles as effectors for chemical threats to skin

氧化脂质和微泡颗粒作为皮肤化学威胁的效应器

基本信息

批准号：
10648739
负责人：
Ravi PRAKASH Sahu
金额：
$ 18.75万
依托单位：
WRIGHT STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-10 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10648739
关键词：
Address Agonist Alkylating Agents Antidotes Antineoplastic Agents Antioxidants Applications Grants Arsenic Arsenicals Binding Burn injury Cell Line Cells Chemical Warfare Chemicals Cisplatin Coupled Cutaneous Cytoplasm Dacarbazine Development Dose Enzymes Epithelial Cells Ethanol Etoposide Exposure to Functional disorder Generations Genetic Goals Human Immunosuppression In Vitro Inflammation Inflammatory Response Injury Ionizing radiation Knockout Mice Knowledge Laboratories Mechlorethamine Mediating Melphalan Membrane Modeling Molecular Morbidity - disease rate Mus Mustard Gas Nitrogen Nuclear Null Lymphocytes Organ Outcome Oxides PUVA Photochemotherapy Pathology Pathway interactions Pilot Projects Platelet Activating Factor Platelet Inhibitors Pre-Clinical Model Process Production Reactive Oxygen Species Receptor Signaling Role Signal Pathway Signal Transduction Site Skin Skin injury Stimulus Techniques Testing Therapeutic Tissues Toxic effect Toxicant exposure Translating Travel UVB induced Ultraviolet B Radiation Vesicants Vesicle acid sphingomyelinase antagonist body system cell injury chemical threat chemotherapeutic agent chemotherapy cigarette smoke cytokine cytotoxic gemcitabine genetic approach in vivo inhibitor insight keratinocyte lipid mediator microvesicles mortality mouse model novel novel strategies oxidized lipid particle pharmacologic platelet activating factor receptor prevent response stressor systemic toxicity tool toxicant transmission process ultraviolet

项目摘要

Abstract The threats to chemical warfare-associated agents, including arsenicals and nitrogen mustards are increasing exceedingly, and no direct antidote is currently available to mitigate the deleterious cutaneous and systemic responses to prevent mortality. Though the associated cytotoxic effects of most of these agents are mediated due to their ability to act as alkylating agents, a significant knowledge gap exits in the understanding of detailed molecular mechanisms of how these vesicants cause cutaneous and systemic toxic effects, and thus, the development of antidotes. The current proposal is built upon the scientific premise that exposure to various pro- oxidative stressors, including alkylating chemotherapeutic agents, and thermal burn injuries (TBI) when coupled with ethanol (ETOH) produce the potent bioactive lipid mediator, Platelet-activating factor (PAF) by both enzymatically and non-enzymatically via reactive oxygen species (ROS). Studies, including ours, have shown that these PAF agonists induce local inflammation, as well as multi-system organ dysfunction (MOD). Importantly, recent studies have indicated that small membrane-bound vesicles known as microvesicle particles (MVP), generated via acid sphingomyelinase (aSMase) enzyme, are released from cells in response to various stressors. These MVP can act as potent signaling agents due to their ability to carry nuclear and cytoplasmic components. More importantly, the current proposal is built upon our discovery that chemotherapeutic agents, and TBI+ETOH via their ability to damage keratinocytes, generate PAF agonists which travel via MVP to induce the local (cutaneous) and systemic responses. Using antioxidants and PAFR-expressing/null cell lines and pharmacologic/genetic inhibition of aSMase enzyme, our studies have implicated the involvement of the PAFR signaling in aSMase activation resulting in PAF-laden MVP release. Based upon these compelling evidences, we hypothesize that chemical warfare-associated agents via their ability to generate ROS, produce PAF agonists and MVP from human and murine keratinocytes in a PAFR-aSMase-dependent manner, which mediate the cutaneous cytotoxic as well as systemic MOD effects. Two aims are proposed to test our hypothesis. Aim 1 will use validated in vitro and ex vivo models and pharmacologic agents to determine the roles of the PAFR, downstream signaling pathways, and aSMase enzyme in chemical warfare-associated agents-induce PAF agonists and MVP generation. Agents to be tested are nitrogen/sulfur mustards and an arsenical. Aim 2 will use PAFR/aSMase-expressing and deficient mouse models, as well as pharmacological inhibitors of PAFR and aSMase to determine the role of PAF-laden MVP generation in the local and systemic responses by topical nitrogen mustargen exposure. Successful completion of this project will (i) fill important mechanistic gaps in and validate novel tools to allow the modulation of PAF-laden MVP generation to mitigate chemical warfare- associated agents’ effects; and ii) address functional roles of PAFR-aSMase in nitrogen mustard-induced local and systemic MOD effects, to provide novel treatment approaches.

抽象的对砷和氮芥等化学战相关制剂的威胁正在增加非常严重，目前没有直接的解毒剂可以减轻对皮肤和全身的有害影响尽管大多数这些药物的相关细胞毒性作用是介导的。由于它们具有充当烷化剂的能力，在详细了解它们方面存在着巨大的知识差距。这些起泡剂如何引起皮肤和全身毒性作用的分子机制，因此，目前的建议是建立在接触各种亲的科学前提之上的。氧化应激源，包括烷基化化疗药物和热烧伤（TBI）与乙醇 (ETOH) 一起产生有效的生物活性脂质介质，血小板激活因子 (PAF) 包括我们在内的研究表明，通过酶促和非酶促活性氧 (ROS)。这些 PAF 激动剂会引起局部炎症以及多系统器官功能障碍 (MOD)。重要的是，最近的研究表明，称为微泡颗粒的小膜结合囊泡 (MVP) 通过酸性鞘磷脂酶 (aSMase) 产生，响应各种环境而从细胞中释放出来这些 MVP 由于具有携带细胞核和细胞质的能力，可以充当有效的信号剂。更重要的是，当前的提议是建立在我们发现化疗药物的基础上的。 TBI+ETOH 通过其损伤角质形成细胞的能力，产生 PAF 激动剂，PAF 激动剂通过 MVP 传播，诱导使用抗氧化剂和 PAFR 表达/无效细胞系进行局部（皮肤）和全身反应。 aSMase 酶的药理学/遗传抑制，我们的研究表明 PAFR 的参与 aSMase 激活中的信号传导导致含有 PAF 的 MVP 释放基于这些令人信服的证据，我们通过化学战相关制剂产生 ROS、产生 PAF 激动剂的能力来追踪它们和 MVP 以 PAFR-aSMase 依赖性方式从人和鼠角质形成细胞中提取，介导提出了两个目标来检验我们的假设。使用经过验证的体外和离体模型和药物制剂来确定 PAFR 的作用，下游信号通路和化学战相关制剂中的 aSMase 酶诱导 PAF 激动剂和 MVP 生成剂是氮/硫芥子气和砷。 PAFR/aSMase 表达和缺陷小鼠模型，以及 PAFR 和 PAFR 的药理学抑制剂 aSMase 通过局部确定负载 PAF 的 MVP 生成在局部和全身反应中的作用氮芥暴露的成功完成该项目将（i）填补重要的机制空白和验证新颖的工具，以允许调制含有 PAF 的 MVP 生成，以减轻化学战- 相关药物的作用；和 ii) 解决 PAFR-aSMase 在氮芥诱导的局部中的功能作用和全身MOD效应，提供新的治疗方法。