Effects of ozone exposure on expression and function of surfactant protein D

臭氧暴露对表面活性蛋白D表达和功能的影响

• 批准号: 7941812
• 负责人: ANGELA HACZKU
• 金额: $ 50万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-28 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7941812
• 关键词: Air Pollutants; Air Pollution; Allergens; Antibodies; Appearance; Area; Asthma; Binding; Biological Assay; Biotin; Bone Marrow; Breathing; C-terminal; CCL17 gene; Carbohydrates; Cell Culture Techniques; Cell surface; Cells; Chronic; Chronic Obstructive Airway Disease; Collagen; Collagen Receptors; Complex; Dendritic Cells; Dependovirus; Disease; Distal; Environmental Exposure; Epithelial; Gene Expression; Genetic; Homeostasis; Human; Immune; Immune system; In Vitro; Individual; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Laboratories; Liquid substance; Lung; Lymphoid; Mediating; Mediator of activation protein; Medical; Mission; Modeling; Morbidity - disease rate; Mus; Myelogenous; N-terminal; NF-kappa B; National Institute of Environmental Health Sciences; Nitrogen; Outcome; Oxygen; Ozone; PTPNS1 gene; Pathology; Patients; Phagocytes; Phase; Post-Translational Protein Processing; Predisposition; Production; Property; Protective Agents; Public Health; Pulmonary Surfactant-Associated Protein D; Receptor Signaling; Regulation; Resolution; Role; Signal Transduction; Structure of parenchyma of lung; TNF gene; Testing; Therapeutic; Thymus Gland; Time; Translational Research; airway hyperresponsiveness; airway inflammation; allergic airway inflammation; asthmatic airway; autocrine; base; calreticulin; chemokine; cytokine; gel electrophoresis; gene replacement; in vivo; knockout gene; lung development; lung injury; lymph nodes; migration; mortality; mutant; novel strategies; oxidative damage; ozone exposure; prevent; public health relevance; receptor binding; response

DESCRIPTION (provided by applicant): This application is submitted in response to the broad challenge area of Translational Science (15-ES-101*) and the specific challenge topic of "Effects of environmental exposures on phenotypic outcomes using non- human models." Chronic inflammatory diseases of the airways such as asthma or chronic obstructive pulmonary disease (COPD) arise from a complex interaction between genetic factors and environmental exposures. Our laboratory has established murine models that mimic the pathology of ozone (O3)-induced exacerbation of airway inflammation. The current application will utilize this model together with gene knockout and conditional over expressor mice to investigate the importance in changes in gene expression and post translational modifications of surfactant protein D, an immunoprotective component of the proximal and distal airway lining. Our studies showed that inhalation of O3, a common air pollutant, induced a significant exacerbation of the asthmatic changes in allergen sensitized mice together with the appearance of abnormal, trimeric SP-D and activated, myeloid dendritic cells. We hypothesize that SP-D acts as a "master switch" between the onset and resolution of airway inflammation: Native SP-D is a suppressor of activation of the proinflammatory dendritic cells and acts through the C-terminal by binding to the negative signal receptor SIRP. In contrast, a de-oligomerized (trimeric) SP-D that has undergone oxidative damage by O3, activates dendritic cells through the exposed N-terminal by ligating the collagen receptor, calreticulin/CD91. The asthmatic airways are particularly susceptible to O3-induced changes in the SP-D molecule because of the presence of mature myeloid dendritic cells carrying the calreticulin/CD91. Aim 1 is to investigate the hypothesis that O3-induced exacerbation of allergic airway inflammation is associated with the appearance of SP-D trimers with an active N-terminal domain. Aim 2 is to study the hypothesis that native SP-D binds to SIRP? and delivers inhibitory signals while trimeric SP-D binds to calreticulin/CD91 and induces proinflammatory activation of NF-kB, TNF and TARC (Ccl17) release by dendritic cells. Aim 3 is to investigate whether the C- terminal suppresses and the trimeric SP-D activates epithelial migration of bone marrow derived, proinflammatory dendritic cells. In vitro bone-marrow derived dendritic cell cultures and in vivo conditional SP-D expressing mice and adeno-associated virus gene replacement of wild-type and mutant SP-D (mimicking either the C-terminal of the molecule or the SP-D trimer) will be studied for maturation and migration of dendritic cells. Results of this project will yield clinically and scientifically significant information on the role of SP-D in regulation of the innate immune system during O3-induced airway changes and on potential therapeutic utilization of this naturally occurring immuno-protective agent in the lung. PUBLIC HEALTH RELEVANCE: Air pollution, particularly O3, induces exacerbations of asthma that substantially worsen morbidity and mortality. In healthy individuals inhalation of O3 elicits mechanisms such as homeostatic increase in SP-D synthesis that protect the lung from development of chronic damage. Currently neither the mechanism of O3-induced SP-D production nor the consequent alterations of the pulmonary immune system is understood. Results from this application will define the implications of SP-D in O3-induced exacerbation of asthma and will provide novel approaches to manipulate the pulmonary immune system.

描述（由申请人提供）：本申请是针对转化科学（15-ES-101*）的广泛挑战领域和“使用非人类模型的环境暴露对表型结果的影响”的特定挑战主题提交的。“呼吸道的慢性炎症性疾病，如哮喘或慢性阻塞性肺病（COPD），是遗传因素和环境暴露之间复杂的相互作用引起的。我们的实验室已经建立了小鼠模型，模拟臭氧（O3）诱导的气道炎症恶化的病理学。本申请将利用该模型连同基因敲除和条件性过表达小鼠来研究表面活性蛋白D的基因表达和翻译后修饰的变化的重要性，表面活性蛋白D是近端和远端气道衬里的免疫保护组分。 我们的研究表明，吸入O3（一种常见的空气污染物）会导致过敏原致敏小鼠的哮喘变化显着加剧，同时出现异常的三聚体SP-D和活化的骨髓树突状细胞。我们假设SP-D在气道炎症的发作和消退之间起“主开关”的作用：天然SP-D是促炎性树突状细胞活化的抑制剂，并通过与负信号受体SIRP结合而通过C-末端起作用。相比之下，经历了O3氧化损伤的去寡聚化（三聚体）SP-D通过连接胶原蛋白受体钙网蛋白/CD 91通过暴露的N-末端激活树突状细胞。 哮喘气道对O3诱导的SP-D分子变化特别敏感，因为存在携带钙网蛋白/CD 91的成熟髓样树突状细胞。目的1是研究假设O3诱导的过敏性气道炎症加重与SP-D三聚体的出现与活性N-末端结构域。 目的二是研究天然SP-D与SIRP结合的假说。 并传递抑制信号，同时三聚体SP-D结合钙网蛋白/CD 91并诱导树突状细胞释放的NF-kB、TNF和TARC（Ccl 17）的促炎活化。目的3是研究C-末端是否抑制和三聚体SP-D激活骨髓来源的促炎树突状细胞的上皮迁移。将研究体外骨髓来源的树突状细胞培养物和体内条件性SP-D表达小鼠以及野生型和突变型SP-D的腺相关病毒基因置换（模拟分子的C末端或SP-D三聚体）对树突状细胞成熟和迁移的影响。该项目的结果将产生有关SP-D在O3诱导的气道变化期间调节先天免疫系统的作用以及这种天然免疫保护剂在肺中的潜在治疗利用的临床和科学重要信息。 公共卫生相关性：空气污染，特别是O3，可诱发哮喘恶化，从而大大加重发病率和死亡率。 在健康个体中，吸入O3可增强机制，如SP-D合成的稳态增加，保护肺免受慢性损伤的发展。目前，O3诱导SP-D产生的机制和随之而来的肺免疫系统的改变都不清楚。本申请的结果将定义SP-D在O3诱导的哮喘急性发作中的意义，并将提供操纵肺免疫系统的新方法。