Nanoparticle Coupled Antioxidants for Respiratory Illness in Veterans

描述（由申请人提供）： 呼吸系统疾病影响了部署到伊拉克或阿富汗的三分之二以上的退伍军人。事实上，呼吸道疾病是持久自由行动（OEF）和伊拉克自由行动（OIF）退伍军人中最常见的医疗条件之一，这是一个一致的发现，影响军事人员，无论年龄，军衔，性别或现役/国民警卫队身份。近23%的OEF/OIF退伍军人认为呼吸系统疾病对部队效力产生了负面影响1。据了解，即使是轻微的上呼吸道感染也会对军事行动产生重大影响，2人们对部署到2011年的人员中呼吸道感染和过敏的频率可能增加表示关切。我们的退伍军人经历了更多的环境暴露，可能有助于这些炎症性呼吸道疾病，包括烟草烟雾和柴油废气。近一半的OEF/OIF退伍军人在部署期间开始或重新开始吸烟。此外，暴露于柴油废气颗粒（DEP）发生在军用车辆和设备中广泛使用柴油发动机。烟草烟雾和DEP都是已知的空气传播毒素，由于自由基而引起广泛的氧化炎症，并可导致过敏性鼻炎恶化3。自由基的快速降解导致活性氧（ROS）4-6。代谢ROS的抗氧化酶因此用作潜在的治疗剂。这些抗氧化剂包括超氧化物歧化酶（SOD），其将超氧化物（最具毒性的ROS之一）转化为活性低得多的过氧化物，以及过氧化氢酶（CAT），其将过氧化物转化为水和氧气。抗氧化剂的治疗用途的一个问题是确保局部施用的药剂保持与呼吸道粘膜接触足够长的时间以有效。正常的粘膜纤毛清除在15-20分钟内将局部药物从呼吸道中清除，此时更希望保持粘膜接触数小时至数天。我们建议使用可生物降解的聚乳酸（PLA）纳米颗粒（NPs）实现SOD和CAT（抗氧化剂）在呼吸道上皮细胞上的靶向递送和保留。我们将同时用SOD和/或CAT和粘蛋白1（MUC 1）的靶向抗体（Ab）包被NP，该抗体与气道上皮细胞的管腔表面结合。我们的初步研究表明，HSNECs在调节对环境刺激的炎症反应中起着关键作用，并且用抗氧化剂靶向HSNECs可以有利地影响随后的APC和T细胞反应。我们的中心假设是OEF/OIF退伍军人经历的烟雾和DEP暴露通过ROS影响HSNEC-APC通讯，并且这种炎症反应可以通过使用靶向呼吸道上皮的抗氧化剂纳米颗粒来改善。具体目标1：确定抗体-抗氧化剂-NP的结合特性和上皮毒性。SOD和CAT将与抗MUC 1 Ab同时共价连接到PLA NP。将制备具有不同比例的SOD、CAT和抗MUC 1的NP，并且将使用HSNEC培养物在体外评估SOD：CAT：抗MUC 1比例对结合特性、功能和毒性的影响。具体目的2：确定抗氧化剂NP在抑制烟雾和DEP诱导的炎症中的体外功效。我们将使用目标1中鉴定的优化的抗MUC 1-SOD/CAT-NP来确定在HSNEC和APC中预防烟雾和DEP诱导的体外炎症反应的功效和能力，作为免疫反应的初始步骤。具体目的3：确定抗氧化剂NP对烟雾和DEP加重的鼻窦炎的体内功效。我们将使用过敏性真菌性鼻窦炎的小鼠模型，通过烟雾或DEP暴露来确定抗MUC 1-SOD/CAT-NP的功效。

DESCRIPTION (provided by applicant): Respiratory illnesses affect over two-thirds of Veterans deployed to Iraq or Afghanistan. The fact that respiratory tract disorders are among the most common medical conditions in Operation Enduring Freedom (OEF) and Operation Iraqi Freedom (OIF) Veterans has been a consistent finding that affects military personnel regardless of age, rank, gender or active duty/National Guard status. Nearly 23% of OEF/OIF Veterans felt unit effectiveness was negatively affected by respiratory illnesses1. Even minor upper respiratory tract infections are known to have a significant impact on military operations2 and concern has been raised about a possible increase in frequency of respiratory tract infections and allergy among personnel deployed to Iraq1. Our Veterans experience an increased number of environmental exposures that likely contribute to these inflammatory respiratory conditions, to include tobacco smoke and diesel exhaust. Nearly half of OEF/OIF Veterans started or re-started smoking during deployment. Additionally, exposure to diesel exhaust particles (DEP) occurs from the extensive use of diesel engines in military vehicles and equipment. Both tobacco smoke and DEP are known airborne toxins that cause extensive oxidative inflammation due to free radicals and can lead to exacerbation of allergic rhinitis3. Quick degradation of free radicals results in reactive oxygen species (ROS)4-6. Antioxidant enzymes that metabolize ROS, thus serve as potential therapeutic agents. These antioxidants include superoxide dismutase (SOD), which converts superoxide, one of the most toxic ROS, to much less reactive peroxide, and catalase (CAT), which converts peroxide into water and oxygen. One problem with therapeutic use of antioxidants is ensuring that topically administered agents remain in contact with the respiratory mucosa long enough to be effective. Normal mucociliary clearance removes topical medications from the respiratory tract within 15-20 minutes when it would be more desirable to maintain mucosal contact for hours to days. We propose to achieve targeted delivery to and retention of SOD and CAT (antioxidants) on respiratory epithelium using biodegradable polylactic acid (PLA) nanoparticles (NPs). We will simultaneously coat NPs with SOD and/or CAT and a targeting antibody (Ab) to mucin 1 (MUC1), which binds to the luminal surface of airway epithelial cells. Our preliminary studies demonstrate that HSNECs play a critical role in modulating the inflammatory response to environmental stimuli, and that targeting HSNECs with antioxidants can favorably impact the subsequent APC and T cell responses. Our central hypothesis is that smoke and DEP exposure experienced by OEF/OIF Veterans impacts HSNEC-APC communication via ROS and that this inflammatory response can be ameliorated through the use of antioxidant nanoparticles targeted at the respiratory epithelium. This hypothesis will be tested by the following specific aims: Specific Aim 1: To determine binding characteristics and epithelial toxicity of antibody-antioxidant- NPs. SOD and CAT will be covalently attached to PLA NPs simultaneously with antiMUC1 Ab. NPs with different ratios of SOD, CAT and antiMUC1 will be prepared and effects of the SOD:CAT:antiMUC1 ratios upon binding characteristics, function and toxicity will be evaluated in vitro using HSNEC cultures. Specific Aim 2: To determine in vitro efficacy of antioxidant NPs in inhibiting smoke and DEP induced inflammation. We will use optimized antiMUC1-SOD/CAT-NPs identified in Aim 1 to determine efficacy and ability to prevent smoke and DEP induced in vitro inflammatory response in HSNECs and APCs as the initial steps in the immune response. Specific Aim 3: To determine in vivo efficacy of antioxidant NPs on smoke and DEP exacerbated rhinosinusitis. We will use a murine model of allergic fungal rhinosinusitis with exacerbation by smoke or DEP exposure to determine the efficacy of antiMUC1-SOD/CAT-NPs.