Extracellular vesicles as mediators of injury in inhaled exposures to toxic chemicals.

项目摘要 芥子气和氯再次成为对军事和平民的潜在威胁。 吸入硫芥（SM）和氯可引起急性肺损伤，可导致呼吸道疾病。 衰竭、多器官功能障碍和死亡。肺毒性导致全身损伤的机制 还不清楚我们对吸入卤素（氯和溴）的研究表明， 神经元损伤我们以前已经确定了循环卤素加合物，形成在肺床 并导致远距离器官损伤和疾病致病性。在我们的吸入性SM损伤模型中，我们 表明循环因子如从肺组织和细胞释放的核酸可以 显著地导致肺损伤，并且清除核酸可以减轻损伤和拯救 从死亡。因此，循环因子对肺和全身损伤至关重要。越来越多的证据 表明几种这样的因子作为外泌体（一种细胞外囊泡（EV））中的货物携带。研究 已经表明EV/外来体可能是致病的。我们的初步数据显示， 从支气管肺泡灌洗液（BALF）的氯暴露大鼠是不同的外泌体货物 从对照动物的BALF中获得。此外，我们证明了从支气管肺泡灌洗液中分离的外泌体， 暴露于CEES（2-氯乙基乙基硫醚，又名：半芥子气）（SM的类似物）的动物，当加入细胞时 在培养物中呈剂量依赖性增加的炎性细胞因子和促凝血基因，重要成分 SM诱导和CEES诱导损伤的发病机制。在我们的体内研究中， 含量与BALF蛋白（肺泡屏障渗漏的标志物）相关。这些研究让我们假设 有毒化学品暴露导致致病性外泌体的释放， 炎症和凝血途径以及阻断它们的生物发生或摄取可以减轻损伤， 防止急性发病率和死亡率。因此，我们将（a）表征来源于大肠杆菌的EV/外泌体。 暴露于Cl 2和CEES的大鼠的BALF和血浆，（B）鉴定来自这些的EV/外来体通过其表达的机制。 毒性化学品暴露的动物引起损伤，和（c）评估来自毒性化学品的EV/外来体的致病性 接触化学品的动物是测试功效或生物发生或吸收抑制剂。这些研究将描述 EV/外来体影响肺损伤/疾病严重程度和消退的机制， 确定外泌体是否可以作为潜在的治疗靶点，以减轻由以下原因引起的器官损伤： 吸入有毒化学物质

Project Summary Sulfur mustard (SM) and chlorine have reemerged as a potential threat to both military and civilian populations. Inhaled exposures to sulfur mustard (SM) and chlorine cause acute lung injury, which can lead to respiratory failure, multiorgan dysfunction and death. Mechanisms by which pulmonary toxicity contributes to systemic injury are not clear. Our studies with inhaled halogens (chlorine and bromine) have demonstrated serious cardiac and neuronal injury. We have previously identified circulating adducts of halogens that form in the pulmonary bed and contribute towards distant organ damage and disease pathogenicity. In our inhaled SM models of injury, we demonstrated that circulating factors such as nucleic acids released from pulmonary tissues and cells can contribute significantly to lung damage and that scavenging the nucleic acids can alleviate injury and rescue from mortality. Therefore, circulating factors are critical to pulmonary and systemic injuries. Growing evidence suggests that several such factors are carried as cargo in exosomes a type of extracellular vesicles (EV). Studies have shown that EV/exosomes could be pathogenic. Our preliminary data shows that the composition of cargoes from the bronchoalveolar lavage fluid (BALF) of chlorine exposed rats are distinct from the exosomal cargo obtained from the BALF of control animals. Further, we demonstrate that exosomes isolated from the BALF of animals exposed to CEES (2-chloroethyl ethylsulfide, aka: half mustard), an analog of SM, when added to cells in culture dose-dependently increased inflammatory cytokines and procoagulation genes, important components in the pathogenesis of SM-induced and CEES-induced injuries. In our in vivo studies the SM BALF exosome content correlated with the BALF protein, a marker of leaky alveolar barrier. These studies led us to hypothesize that toxic chemical exposures result in the release of pathogenic exosomes that causes activation of the inflammatory and coagulation pathways and that blocking their biogenesis or uptake can mitigate injury and protect from acute morbidity and mortality. Accordingly, we will (a) characterize EV/exosomes derived from the BALF and plasma of rats exposed to Cl2 and CEES, (b) identify mechanisms by which EV/exosomes from these toxic chemical-exposed animals cause injury, and (c) evaluate pathogenicity of EV/exosomes derived from toxic chemical-exposed animals are test efficacy or biogenesis or uptake inhibitors. These studies will delineate mechanisms by which EV/exosomes influence pulmonary injury/disease severity and resolution and also determine whether exosomes can serve as potential therapeutic targets in mitigating organ injury caused by inhaled toxic chemicals.