Down-regulation of oxidant-induced airway inflammation though modulation of NRF2

描述(由申请者提供)：本提案概述了为期3年的K23指导下的以患者为导向的研究职业发展奖(K23)，旨在完善候选人作为内科科学家的培训，并为专注于开发抗呼吸道氧化应激疗法的多学科、转化性研究计划做准备。这项建议的目标是在候选人将基础科学和临床研究的经验应用于开发独立的转化性研究项目时，在药物发现/开发方面提供支持、培训，并在基于干预的概念验证研究方面提供指导。最近来自该小组和其他人的证据强调了细胞内抗氧化酶在加重臭氧诱导的呼吸道炎症中的作用。缺乏抗氧化酶谷胱甘肽S转移酶MU1的健康志愿者在舱内暴露于百万分之0.4的臭氧后，中性粒细胞炎症增加。GSTM1和许多其他II相抗氧化酶(NQO1、GSTP1、HO-1)受主要转录因子NF-E2相关因子2(NRF2)的调节。小鼠模型和对慢性阻塞性肺疾病患者的研究表明，NRF2的缺陷与氧化剂介导的肺损伤有关。因此，NRF2是一个强有力的候选者，可以调节对无处不在的吸入氧化剂如臭氧引起的呼吸道炎症的保护作用。这里的目的是使用萝卜硫素(SFN)，一种从特殊培育的西兰花中提取的抗氧化化合物，被发现可以上调NRF2和NRF2调节的II相酶(GSTM1、GSTP1、HO1和NQO1)的表达，以检验口服SFN是否可以减少正常志愿者中由臭氧诱导的呼吸道炎症。其次，来自轻、中度持续性过敏性哮喘患者的已分化鼻腔上皮细胞将被用来检测SFN对这些上皮细胞的治疗是否改变了臭氧诱导的炎症反应。应聘者将通过拟议的教学课程获得药物发现过程中的经验，这些药物可以针对氧化应激过程，并在指导团队的协助下完成这些目标。指导团队由三位在指导年轻科学家和开发翻译研究项目方面拥有丰富经验的人员组成：David Peden博士，翻译科学家，具有环境污染、氧化应激和首席导师方面的专业知识；Wesley Burks博士，翻译研究员，专注于针对食物过敏的I/II期临床试验干预；以及Angela Kashuba博士，北卡罗来纳大学临床药理学研究员，在临床药代动力学和药效学研究方面拥有丰富经验。 与公共卫生相关：臭氧引起的呼吸道并发症在美国和世界范围内是一个重大的公共卫生负担。尽管有令人信服的证据表明，肺部氧化损伤会导致全身有害的健康后果，但环境引起的氧化应激的治疗选择有限，在改善儿童、老年人和过敏性哮喘患者等易感人群的健康方面没有取得重大变化。这项研究有可能确定一条可以作为预防或治疗易感人群因吸入环境氧化剂引起的呼吸道炎症的靶点的途径。鉴于哮喘等呼吸道疾病给公众健康造成的巨大负担，以及缺乏针对氧化剂引起的呼吸道炎症的靶向治疗，潜在的影响是巨大的。

DESCRIPTION (provided by applicant): This proposal outlines a 3 year K23 Mentored Patient-Oriented Research Career Development Award (K23) designed to refine the candidate's training as a physician scientist and to prepare for a multidisciplinary, translational research program focused on the development of therapies against airway oxidative stress. The goal of this proposal is to provide support, training in drug discovery/development, and guidance in intervention-based proof of concept studies as the candidate applies experience in basic science and clinical research to the development of an independent translational research project. Recent evidence from this group and others has emphasized the role of intracellular antioxidant enzymes in exacerbation of O3-induced airway inflammation. Healthy volunteers lacking the antioxidant enzyme, Glutathione S Transferase Mu 1 (GSTM1), suffered from increased neutrophilic airway inflammation after chamber exposure to 0.4 parts per million (ppm) ozone (O3). GSTM1 and numerous other phase II antioxidant enzymes (NQO1, GSTP1, HO-1) are regulated by the master transcription factor NF-E2- related factor 2 (NRF2). Murine models and studies of patients with chronic obstructive pulmonary disease suggest that defects in NRF2 are associated with oxidant-mediated lung injury. Therefore, NRF2 is a strong candidate to modulate in protection against airway inflammation caused by ubiquitous inhaled oxidants such as O3. The intent here is to use sulforaphane (SFN), an antioxidant compound derived from specially bred broccoli that was found to upregulate expression of NRF2 and NRF2-regulated Phase II enzymes (GSTM1, GSTP1, HO1, and NQO1), to examine if NRF2 induction with oral SFN supplementation will reduce O3-induced airway inflammation in normal volunteers. Second, cultured differentiated nasal epithelial cells derived from mild-moderate persistent allergic asthmatics will be used to examine if SFN treatment of these epithelial cells modifies O3-induced inflammatory responses. The candidate will acquire experience in the drug discovery process of pharmacologic agents that can target oxidative stress processes through the proposed didactic coursework as well as with the completion of these aims with the assistance of the mentoring team. The mentoring team consists of three individuals with extensive experience in mentoring young scientists and in developing translational research programs: Dr. David Peden, a translational scientist with expertise on environmental pollution, oxidative stress, and lead mentor; Dr. Wesley Burks, a translational researcher focused on phase I/II clinical trial interventions against food allergy; and Dr. Angela Kashuba, the director f the clinical pharmacology fellowship at UNC with extensive experience in clinical pharmacokinetic and pharmacodynamic studies. PUBLIC HEALTH RELEVANCE: Respiratory complications caused by O3 represent a significant public health burden in the United States and worldwide. Despite compelling evidence that pulmonary oxidative insults lead to systemic deleterious health outcomes, treatment options for environmentally-induced oxidative stress are limited and no significant changes have been made in improving the health of susceptible populations such as children, the elderly and allergic asthmatics. This research has the potential to identify a pathway that could be targeted by prevention or treatment of respiratory inflammation caused by exposure to inhaled environmental oxidants in susceptible individuals. Given the significant public health burden of respiratory diseases such as asthma and the lack of targeted therapeutics for oxidant-induced airway inflammation, the potential impact is great.