Cellular interactions in asthma

描述（由申请人提供） 摘要：哮喘是一种肺部疾病，它会导致气道收缩，以应对花粉，霉菌，病毒感染，甚至冷空气等触发因素。在严重的情况下，哮喘的气道阻塞可能是致命的。哮喘特别影响大量儿童，并且该疾病在全世界范围内的流行率正在增长。在大多数哮喘患者中，存在炎性细胞对肺部的慢性浸润。这些炎性细胞已被证明对疾病病理学做出重要贡献，但目前对该疾病的治疗是不够的。 在确定哮喘患者肺部积聚的特定类型的炎性细胞方面已经取得了相当大的进展。大量的研究已经针对通过基因消融或通过药理学试剂抑制特定细胞相关分子来测试这些单个细胞类型的作用，然后测量对肺功能和炎症的影响。然而，虽然这种实验方法已经确定了构成哮喘之谜的许多关键部分，但它未能提供这些部分如何组合在一起导致疾病的图片。事实上，我们对体内导致早期过敏性致敏或随后的慢性哮喘的细胞-细胞相互作用知之甚少。 本研究提出应用尖端的显微镜技术来表征哮喘小鼠模型中的细胞相互作用。这项研究的第一步将是确定通过双光子显微镜观察肺部炎症细胞的方法。感兴趣的特定细胞类型是肥大细胞、嗜碱性粒细胞、嗜酸性粒细胞、嗜中性粒细胞、巨噬细胞、单核细胞、树突细胞、T细胞和B细胞。这些细胞将通过以谱系特异性方式表达荧光蛋白而特异性地可视化，或 用荧光染料或抗体标记。在某些情况下，这项研究将能够利用现有的转基因小鼠品系来实现这一目的。本研究的第二步将是通过双光子显微镜来表征这些细胞之间的相互作用。首先将确定相互作用的特定细胞类型，然后将评估这些接触的频率和持续时间。这些接触的性质将通过遗传和药理学扰动来探测。本研究的第三步是确定这些细胞相互作用在肺内的位置，并确定促进这些相互作用的组织微环境的特征。一个特别的重点领域将是炎症细胞与肺的基质成分，包括上皮细胞和平滑肌细胞的相互作用。总之，这项研究将为炎症细胞如何相互沟通以及肺部驻留细胞如何导致哮喘发病机制提供新的见解。这些发现可能为理解哮喘提供一个新的范例，最终可能导致新的治疗干预措施。 公共卫生相关性：这项研究旨在提高我们对炎症如何导致哮喘的理解。这项研究的总体目标是确定肺部哪些炎性细胞相互作用，以及它们如何沟通导致疾病。这项工作的结果可能为针对这些细胞相互作用的新型哮喘治疗提供基础。

DESCRIPTION (Provided by the applicant) Abstract: Asthma is a lung disease that causes constriction of the airways in response to triggers such as pollen, mold, viral infection, or even cold air. In severe cases, the obstruction f the airways in asthma can be fatal. Asthma particularly affects a large number of children and the disease is growing in prevalence worldwide. In the majority of asthma patients, chronic infiltration of the lungs with inflammatory cells is present. These inflammatory cells have been shown to make an important contribution to disease pathology, yet current treatments for the disease are inadequate. Considerable progress has been made in defining the specific types of inflammatory cells that accumulate in the lungs of asthma patients. A large number of studies have been directed at testing the roles of these individual cell types by genetic ablation or the inhibition of particular cell-associated molecules by pharmacological agents, followed by measurement of the effects on lung function and inflammation. However, while this experimental approach has identified many of the key pieces that make up the asthma puzzle, it has failed to provide a picture of how these pieces fit together to cause disease. Indeed, little is known about the cell-cell interactions in vivo that lead to early allergic sensitization or subsequent chronic asthma. This study proposes to apply cutting-edge microscopy techniques to the characterization of cellular interactions in asthma using mouse models. The first step of this study will be to identify methods to visualize inflammatory cells in the lungs by two-photon microscopy. The particular cell types of interest will be mast cells, basophils, eosinophils, neutrophils, macrophages, monocytes, dendritic cells, T cells, and B cells. These cells will be specifically visualized by the expression of fluorescent proteins in a lineage-specific fashion, or by labeling with fluorescent dyes or antibodies. In several cases, this study will be able to draw upon existing lines of transgenic mice for this purpose. The second step of this study will then be to characterize the interactions among these cells by two-photon microscopy. The particular cell types that interact with each other will first be identified, and then the frequency and duration of these contacts will be evaluated. The nature of these contacts will then be probed by genetic and pharmacological perturbations. The third step in this study will be to identify the locations of these cellular interactions within the lung and define the features of the tissue microenvironment that promote these interactions. A particular area of emphasis will be the interaction of inflammatory cells with stromal components of the lung, including epithelial cells and smooth muscle cells. Taken together, this study will provide new insights into how inflammatory cells communicate with each other and lung-resident cells leading to asthma pathogenesis. These findings are likely to provide a new paradigm for the understanding of asthma, which may ultimately lead to novel therapeutic interventions. Public Health Relevance: This study aims to improve our understanding of how inflammation contributes to asthma. The overall goal of this study is to determine which inflammatory cells in the lungs interact with each other and how they communicate to cause the disease. The results of this work may provide a foundation for new types of asthma therapeutics that target these cellular interactions.