Proteomics of flavorings-induced airway disease

调味品引起的气道疾病的蛋白质组学

• 批准号: 8700968
• 负责人: Matthew Wolf Foster
• 金额: $ 23.35万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8700968
• 关键词: Proteomics; flavorings; induced; airway; disease

DESCRIPTION (provided by applicant): The overall goal of this application is to improve the diagnosis and treatment of bronchiolitis obliterans (BO) that occurs in the workplace as a result of exposure to artificial flavors. Workers in the food manufacturing industry are at significant rik for occupational airway disease due to exposure to commonly used artificial flavorings. In particular, diacetyl (DA) and pentanedione (PD), components used in butter flavoring, have been linked to the development of bronchiolitis obliterans (BO), an irreversible airway fibrosis. As a result, NIOSH NORA objectives include strategic goals focused on work-related airway diseases, specifically targeting studies of "diacetyl and other potentially harmful artificial flavorings" to improve workplace risk assessment (Goal 5.2.2) and define mechanisms of toxicity (Goal 5.1.3). This proposal directly addresses the NIOSH objectives for the Manufacturing Sector and Respiratory Disease Cross-sector. It is also consistent with the research-to-practice (r2p) goals by targeting an area prioritized as a major occupational health issue facing manufacturing workers. A primary goal of this project is to identify novel biomarkers of early lung injury after exposure to the artificial flavors DA and PD. Our overall hypothesis is that the protein signature from normal human bronchial epithelial (NHBE) cells exposed to DA or PD will provide a useful discovery platform for human biomarker development. By subsequently validating the most overexpressed proteins in a pre-clinical rodent BO model, we identify those targets that can best be translated into relevant blood or sputum tests for risk assessment in chemical flavoring exposed workers. As a secondary goal, we will use bioinformatic pathway analysis of the protein expression pattern of the NHBE cells in response to DA or PD to better define mechanisms of flavoring cellular toxicity, and determine which cellular pathways are activated in response to these agents. In preliminary studies in support of this application we have established that treatment of NHBE with DA or PD in concentrations up to 40 mm results in production of the cytokine interleukin (IL)-8 in a dose dependent manner, with minimal effects on cell viability. We demonstrated parallel increases in IL-8 in the lung flui in DA or PD-induced in vivo models of occupational BO in rodents. In the current application we will extend this preliminary data and 1) quantify the full spectrum of secreted proteins from cultured NHBE in response to DA or PD using state-of-the-art proteomic analysis technology, and apply bioinformatics analysis to determine pathways dysregulated by flavoring chemicals and 2) measure identified candidate proteins in the blood and lung fluid DA or PD-induced rodent models of occupational BO to prioritize selection of biomarkers for workplace studies. Our proposed unbiased discovery based approach is likely to deliver novel biomarkers that could improve early recognition of risk in the workplace and to provide new insights into the mechanisms of chemical flavoring toxicity that could suggest new actionable targets for BO prevention or treatment.

描述（由申请人提供）：本申请的总体目标是改善因人造口味而导致在工作场所中发生的细支气管炎的诊断和治疗。食品制造业的工人因接触常用的人造调味剂而对职业气道疾病的重大RIK处于重要状态。特别是，在黄油调味剂中使用的成分二乙酰基（DA）和五丹二酮（PD）与不可逆的气道纤维化（BO）（BO）（BO）的发展有关。结果，NIOSH NORA目标包括针对与工作相关的气道疾病的战略目标，特别是针对“二乙酰基和其他潜在有害的人工调味剂”的研究，以改善工作场所的风险评估（目标5.2.2）和固定毒性机制（目标5.1.3）。该提案直接解决了制造业和呼吸系统疾病跨部门的NIOSH目标。这也与练习对目标（R2P）的目标一致，该目标是针对优先作为制造工人面临的主要职业健康问题的领域。该项目的主要目的是鉴定暴露于人造口味DA和PD后的早期肺损伤的新型生物标志物。我们的总体假设是，暴露于DA或PD的正常人支气管上皮（NHBE）细胞的蛋白质特征将为人类生物标志物发育提供有用的发现平台。通过随后验证临床前啮齿动物BO模型中最过表达的蛋白质，我们确定了那些最能将其转化为相关的血液或痰液测试的靶标，以评估化学调味剂暴露的工人的风险评估。作为次要目标，我们将使用对DA或PD响应NHBE细胞的蛋白质表达模式的生物信息学途径分析，以更好地定义调味细胞毒性的机制，并确定哪些细胞途径响应这些药物而激活。在支持此应用的初步研究中，我们确定以高达40 mM的DA或PD处理NHBE，以剂量依赖性的方式导致细胞因子白介素（IL）-8的产生，对细胞生存能力的影响很小。我们证明了DA或pd诱导的啮齿动物的体内模型中的肺氟中IL-8的平行增加。 In the current application we will extend this preliminary data and 1) quantify the full spectrum of secreted proteins from cultured NHBE in response to DA or PD using state-of-the-art proteomic analysis technology, and apply bioinformatics analysis to determine pathways dysregulated by flavoring chemicals and 2) measure identified candidate proteins in the blood and lung fluid DA or PD-induced rodent models of occupational BO to prioritize selection of工作场所研究的生物标志物。我们提出的基于发现的基于发现的方法可能会提供新颖的生物标志物，这些标志物可以提高工作场所中风险的早期认识，并提供有关化学调味毒物毒性机制的新见解，这可能暗示了可行​​的预防或治疗的新目标。