Molecular Characterization Core

项目总结 肺气肿和肺纤维化是肺部对长期吸烟暴露的两种反应，两者都是 这可能受线粒体功能障碍和几丁质酶途径的影响。基因表达谱有 成功识别与特发性肺纤维化(IPF)有关的多个基因；基因表达研究 在慢性阻塞性肺疾病(COPD)方面有更多的变数结果。细胞的光谱 肺中的异质性现在可以通过单细胞RNA测序来分析，这已经在 在人类IPF和最近在人类COPD肺组织中进行的研究，作为IPF研究中的比较。要支持所有 三个项目，分子表征核心将使用单细胞和标准RNA测序来测试 COPD和IPF的细胞类型、细胞亚群和细胞类型特异性基因表达变化的差异 在这个PPG收集的人类样本和将在项目1和2中使用的小鼠模型中。我们 将解决以下具体目标：(1)我们将在肺组织中进行单细胞RNA测序 博莱霉素和香烟烟雾暴露小鼠模型以确定与肺相关的细胞变化 纤维化和空域扩大。我们将定位ALL相关基因的细胞表达 三个项目。(2)我们将对来自香烟烟雾的小鼠肺中的单细胞RNA进行测序 用于识别与合并的空域扩大和气道纤维化相关的细胞变化的模型 模特。我们将定位所有三个项目相关基因的细胞表达。(3)我们将批量执行和 支气管镜下采集的呼吸道上皮细胞和肺泡巨噬细胞的单细胞RNA测序 临床生物信息库核心。我们将确定细胞亚群和基因表达的差异 COPD、IPF和控制吸烟者。这个新的核心将与关于小鼠的项目1和2密切互动 模型和用于人体研究的项目3和核心C(临床生物库)。关于大尺度的分析 在这个核心中产生的数据将需要与核心B(呼吸计算发现)合作。

PROJECT SUMMARY Emphysema and pulmonary fibrosis are two responses of the lung to chronic cigarette smoke exposure, both of which may be affected by mitochondrial dysfunction and chitinase pathways. Gene expression profiling has successfully identified numerous genes involved in idiopathic pulmonary fibrosis (IPF); gene expression studies in chronic obstructive pulmonary disease (COPD) have had more variable results. The spectrum of cellular heterogeneity in the lung can now be assayed by single cell RNA sequencing, which has been performed in human IPF and recently in human COPD lung tissue, serving as a comparison in an IPF study. To support all three projects, the Molecular Characterization core will use single cell and standard RNA sequencing to test for differences in cell types, cell subpopulations, and cell-type specific gene expression changes in COPD and IPF in human samples collected in this PPG and in the mouse models which will be used in Projects 1 and 2. We will address the following Specific Aims: (1) We will perform single cell RNA-sequencing in lung tissues from murine models of bleomycin and cigarette smoke exposure to identify cellular changes associated with lung fibrosis and airspace enlargement, respectively. We will localize cellular expression of relevant genes from all three projects. (2) We will perform single cell RNA-sequencing in mouse lungs from the cigarette smoke-poly I:C model to identify cellular changes associated with the combined airspace enlargement and airway fibrosis in this model. We will localize cellular expression of relevant genes from all three projects. (3) We will perform bulk and single cell RNA-sequencing on airway epithelial cells and alveolar macrophages collected at bronchoscopy in the Clinical Biorepository Core. We will identify cell subpopulations and gene expression differences between COPD, IPF, and control smokers. This new core will interact closely with Projects 1 and 2 regarding the murine models and with Project 3 and Core C (Clinical Biorepository) for the human studies. Analysis of the large-scale data generated in this core will require collaboration with Core B (Respiratory Computational Discovery).