Molecular Characterization Core

项目摘要 肺气肿和肺纤维化是肺对慢性香烟烟雾暴露的两种反应， 其可能受到线粒体功能障碍和几丁质酶途径的影响。基因表达谱分析 成功鉴定了许多参与特发性肺纤维化（IPF）的基因;基因表达研究 在慢性阻塞性肺疾病（COPD）中，有更多的可变结果。细胞的光谱 肺中的异质性现在可以通过单细胞RNA测序来测定，该测序已经在 人IPF和最近在人COPD肺组织中进行的研究，用作IPF研究中的比较。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中细胞类型、细胞亚群和细胞类型特异性基因表达变化的差异 在本PPG中采集的人体样本以及将用于项目1和2的小鼠模型中。我们 我们将致力于以下具体目标：（1）我们将在来自以下的肺组织中进行单细胞RNA测序： 博莱霉素和香烟烟雾暴露的小鼠模型，以鉴定与肺相关的细胞变化 纤维化和气腔扩大。我们将定位相关基因的细胞表达， 三个项目。(2)我们将在香烟烟雾中的小鼠肺中进行单细胞RNA测序-聚I：C 该模型用于识别与该患者中的合并的空域扩大和气道纤维化相关的细胞变化。 模型我们将定位所有三个项目相关基因的细胞表达。(3)我们将进行批量生产， 在支气管镜检查中收集的气道上皮细胞和肺泡巨噬细胞的单细胞RNA测序， 临床生物储存中心我们将确定细胞亚群和基因表达的差异， COPD、IPF和对照吸烟者。这个新的核心将与项目1和项目2密切互动， 模型，并与项目3和核心C（临床生物储存库）用于人体研究。大规模分析 在此核心中生成的数据将需要与核心B（呼吸计算发现）协作。

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).