Molecular Characterization Core

项目概要 肺气肿和肺纤维化是肺部对长期接触香烟烟雾的两种反应，两者都是 这可能受到线粒体功能障碍和几丁质酶途径的影响。基因表达谱有 成功鉴定了许多与特发性肺纤维化（IPF）有关的基因；基因表达研究 在慢性阻塞性肺疾病（COPD）中的研究取得了更多不同的结果。蜂窝频谱 现在可以通过单细胞 RNA 测序来分析肺部的异质性，该测序已在 人类 IPF 和最近在人类 COPD 肺组织中的研究，作为 IPF 研究中的比较。为了支持所有 三个项目中，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 中收集的人类样本以及将在项目 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 博来霉素和香烟烟雾暴露的小鼠模型，用于识别与肺相关的细胞变化 分别为纤维化和空腔扩大。我们将定位所有相关基因的细胞表达 三个项目。 (2) 我们将对香烟烟雾-poly I:C 的小鼠肺部进行单细胞 RNA 测序 模型来识别与联合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中相关基因的细胞表达。 (3) 我们将进行批量和 对支气管镜检查收集的气道上皮细胞和肺泡巨噬细胞进行单细胞 RNA 测序 临床生物样本库核心。我们将识别细胞亚群和基因表达差异 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 模型以及用于人类研究的项目 3 和核心 C（临床生物库）。大规模分析 该核心生成的数据需要与核心 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).