Regulation of Ozone Induced Neutrophilic Lung Inflammation by ATP Synthase Complex V

炎症是对宿主环境的物理或化学扰动的一种保护性反应。臭氧是一种有毒的环境污染物，可导致中性粒细胞肺部炎症和损伤。虽然肺部通常有中性粒细胞驻留，但肺内的暴发性中性粒细胞反应与肺泡毛细血管屏障的破坏和肺功能受损有关。中性粒细胞募集的基本机制及其在臭氧诱导的肺部炎症中的作用尚不完全清楚。因此，我的研究的主要目的是利用一系列体外、体内和体内的细胞和分子方法来了解中性粒细胞激活的基本机制及其在肺部炎症中的作用。我最近开发了一个复杂的低剂量(0.05ppm)臭氧诱导C57BL/6NJ品系小鼠肺部炎症的模型，并对其进行了表征。这种臭氧诱导的肺部炎症的特征是显著的中性粒细胞和嗜酸性粒细胞死亡，形成核细胞外陷阱(Net)，血小板聚集和肺泡收缩减少。我们对三磷酸腺苷合成酶复合体V(ATPS-V)在臭氧暴露小鼠肺血管和肺泡间隔中的表达进行了新的观察。但ATPS-V在肺部炎症，特别是中性粒细胞迁移和肺泡通透性中的基础作用尚不清楚。考虑到已有证据表明ATPS-V在中性粒细胞迁移中的作用，我的短期目标是利用这个模型来解决以下基本问题：1.ATPS-V是否促进臭氧诱导的急性肺中性粒细胞迁移和激活？2.ATPS-V是否恢复臭氧诱导的肺泡收缩动力学？3.ATPS-V是否保护臭氧诱导的肺泡屏障受损？我将利用我的专业知识，使用先进的方法，如显微镜使能细胞表型，计算机断层扫描(CT)，同步辐射X射线多图像照相(MIR)，活体肺成像和微正电子发射断层扫描(PET-CT)来解决这些基本问题。目前的拨款建议使用跨学科的自然科学，以揭示与空气一起吸入的臭氧的基本影响。从拟议的实验中获得的数据将加深我们对低剂量臭氧暴露引起的肺部炎症机制的理解。这些发现将阐明迄今未知的ATPS-V在肺部炎症中的作用，特别是中性粒细胞迁移、肺泡毛细血管通透性和肺泡收缩功能。这些研究将进一步为器官生理学的研究提供新的成像和活体工具。在某个时候，这些基本数据可能会影响加拿大的环境政策。最后，在加拿大唯一的同步加速器工厂，使用先进和独特的成像工具为HQP培训做出了贡献。

Inflammation is a protective response against physical or chemical perturbation of the host environment. Ozone is a toxic environmental pollutant and is attributed to cause neutrophilic lung inflammation and injury. Although lungs are usually patrolled by resident neutrophils, a fulminant neutrophil response in the lungs is associated with disruption of the alveolar capillary barrier and impaired lung function. The fundamental mechanisms of neutrophil recruitment and their role in ozone-induced lung inflammation are not fully understood. Thus, the primary thrust of my research is to understand the fundamental mechanisms of neutrophil activation and their role in lung inflammation using an array of in vitro, intravital and in vivo cell and molecular methods. I have recently developed and characterized a sophisticated model of lung inflammation induced with low dose (0.05 ppm) ozone in C57BL/6NJ strain mice. This ozone-induced lung inflammation is characterized by marked neutrophil and eosinophil cell death, formation of nuclear extracellular traps (NETs), platelet clumping and reduced alveolar contraction. We made a novel observation of expression of the ß subunit of ATP synthase complex V (ATPS-V) in the lung vasculature and alveolar septa of ozone-exposed mice. But the fundamental role of ATPS-V in lung inflammation specifically neutrophil migration and alveolar permeability is not known. Considering that there is previous evidence of role of ATPS-V in neutrophil migration, my short term objectives are to use this model to address the following fundamental questions: 1. Does ATPS-V enhance ozone induced acute lung neutrophil migration and activation? 2. Does ATPS-V recover ozone induced alveolar contractile dynamics? 3. Does ATPS-V protect against ozone induced compromised alveolar barrier? I will use my expertise in the use of advanced methods such as microscopy enabled cell phenotyping, computed tomography (CT), synchrotron x-ray multiple image radiography (MIR), intravital lung imaging and micro positron emission tomography (PET-CT) to address these fundamental questions. The current grant proposes the utilization of cross-disciplinary natural sciences in order to reveal fundamental effects of ozone inhaled with the air. The data obtained from the proposed experiments will enhance our understanding of mechanisms of lung inflammation induced with low dose exposure to ozone. The findings will elaborate on the hitherto unknown role of ATPS-V in lung inflammation specifically neutrophil migration, alveolar capillary permeability and alveolar contractile function. The studies will further provide new imaging and in vivo tools to study organ physiology. The fundamental data may, at some point, influence environmental policies in Canada. Lastly, there is contribution to the HQP training using advanced and unique imaging tools at Canada's only Synchrotron facility.