Identifying Molecular Phenotype of Normal and Asthmatic Bronchial Smooth Muscle

哮喘是一种常见病，有几种有效的治疗方法，包括吸入 类固醇和兴奋剂。尽管如此，一小群患者具有严重的 与气道重塑相关的持续病程。气道的一个重要特征 重塑是支气管平滑肌（BSM）表型的改变， 典型的特征是细胞数量和大小的扩张，以及超 对特异性和非特异性激动剂的反应性。虽然哮喘的基础 发病机制及其临床表现，缺乏有关基础知识， 错乱的BSM表型是该领域中正在进行的、未解决的问题。这是 表现为缺乏关于潜在的分子信号的信息， 支气管高反应性和缺乏专门针对逆转 哮喘BSM表型。造成这种状况的一个主要因素是， 缺乏支持高保真分离纯BSM细胞的工具/方法， 哮喘肺进行分析。为了克服这一点，我们开发了一种独特的转基因 BSM单独表达绿色荧光蛋白（hrGFP）小鼠， 血管平滑肌表达绿色荧光蛋白（hrGFP）和红色荧光蛋白（dsRed）; 从而第一次提供了一种可靠的方法， 两个平滑肌细胞群，以及来自其他肺细胞， 流式细胞仪使用这种独特的小鼠，我们的计划是进行全面的mRNA 和BSM RNA的miRNA谱分析，以测试以下广泛基础的假设：1）BSM 表达不同的基因签名，2）该签名的改变介导 哮喘BSM表型。我们的计划是使用分析数据来生成 正常人和哮喘患者中完整和差异表达的mRNA和miRNA BSM。去调控的miRNAs、mRNA表达与肿瘤细胞特性之间的关系 哮喘BSM中的活性信号通路将通过生物信息学和 功能研究。在这项工作结束时，我们将启动一个进程， 标志着哮喘领域的知识空白，并将建立一个基础 未来的各种研究。

Asthma is a common disease with several effective treatments, including inhaled steroids and ¿-agonists. Despite this, a small subpopulation of patients has a severe unrelenting course associated with airway remodeling. A central feature of airway remodeling is alteration in bronchial smooth muscle (BSM) phenotype, which is classically characterized by expansion of cell number and size, and increased hyper- reactivity to specific and non-specific agonists. While fundamental to asthma pathogenesis and its clinical manifestations, a lack of knowledge regarding the basis for a deranged BSM phenotype is an ongoing, unresolved, issue in the field. This is manifested by the paucity of information regarding the molecular signals underlying bronchial hyper-reactivity and by the lack of treatments directed specifically at reversing the asthmatic BSM phenotype. One major contributing factor to this state-of-affairs is the lack of tools/methodologies that support the high fidelity isolation of pure BSM cells from asthmatic lungs for analysis. To overcome this, we developed a unique transgenic mouse in which BSM singly express a green fluorescent protein (hrGFP) whereas vascular smooth muscle express green (hrGFP) and red fluorescent proteins (dsRed); thereby providing for the first time a reliable methodology for separating each of these two smooth muscle cell populations from one another, and from other lung cells using flow cytometry. Using this unique mouse, our plan is to perform comprehensive mRNA and miRNA profiling of BSM RNA to test the following broad based hypothesis: 1) BSM express a distinct genetic signature and 2) alterations in this signature mediate asthmatic BSM phenotypes. Our plan is to use the profiling data to generate lists of complete and differentially expressed mRNAs and miRNAs in normal and asthmatic BSM. Relationships between deregulated miRNAs, mRNA expression, and the identity of active signaling pathways in asthmatic BSM will be examined by bioinformatic and functional studies. At the end of this work, we will have initiated a process to fill a marked knowledge void in the asthma field and will have established a foundation for a variety of future studies.