Progenitors, Mechanisms of Differentiation, and Functions of Lung M Cells

肺 M 细胞的祖细胞、分化机制和功能

基本信息

批准号：
10673927
负责人：
JAYARAJ RAJAGOPAL
金额：
$ 79.52万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10673927
关键词：
Ablation Antigen Presentation Antigens Applications Grants Attention B-Lymphocytes Biology Bronchus-Associated Lymphoid Tissue Cell Communication Cell Culture Techniques Cell Differentiation process Cell Lineage Cell physiology Cells Data Data Set Dendritic Cells Developmental Biology Disease Disease model Epithelial Cells Epithelium Event Foundations Gastrointestinal tract structure Genetic Human Immune Immunologics Infectious Lung Disorder Inflammation Inflammatory Influenza Label Lipopolysaccharides Literature Lung Lung infections Lymphoid Follicle M cell Maintenance Modeling Molecular Mucosal Immunity Mucositis Mucous Membrane Mus Neuroendocrine Cell Organ Photons Physiological Pulmonary Inflammation Reagent Reporting Role SOX8 gene Sampling Signal Pathway Signal Transduction Slice Stimulus TNFSF11 gene Testing Time Trachea Viral Virus Diseases airway epithelium airway inflammation cell type chemokine cytokine design directed differentiation imaging platform immunohistochemical markers influenza infection mouse model new therapeutic target novel pathogen pharmacologic progenitor pulmonary function stem cell biology stem cells tool transcriptome transcytosis

项目摘要

M cells have been shown to sample luminal antigens and use transcytosis to deliver these antigens to underlying antigen presenting dendritic cells. Thus, they serve as key regulators of mucosal immunity. Despite the vast body of literature on the immunologic role of M cells in other mucosal organs, airway M cells have only recently been described and almost no functional or molecular characterization has been performed. We present the first single cell transcriptomes of murine airway M cells and identify airway M cell signaling cascades that regulate inflammation. We have discovered that airway M cells occur as solitary cells in the murine trachea and as patches in the murine small airway, and we report the directed differentiation of human airway M cells from primary human airway epithelium. Finally, we demonstrate that airway M cells are induced by the administration of the signaling factor RANKL, treatment with Lipopolysaccharide (LPS), and influenza infection. Interestingly, M cells can occur as solitary cells, but in the setting of physiologic stimuli like LPS or influenza infection, patches of M cells are associated with lymphoid follicles suggesting a functional epithelial-immune unit. In this grant application, we propose to use our combined expertise in stem cell biology, epithelial biology, lung inflammation, and influenza infection to define the functional biology of lung M cells. We will start by defining the cellular origins of murine and human airway M cells. We have now generated a comprehensive battery of murine genetic driver lines for lineage tracing all the cells of airway epithelium. Herein, we propose to deploy this set of murine lines, for the first time, in order to precisely define all the putative parental epithelial cell types that can give rise to M cells. We will also clarify the role of RANKL-RANK signaling pathways in murine models of inflammation. Finally, to define airway M cell functions, we constructed a new Sox8-CreER driver mouse that allows us to specifically label, genetically modify, and ablate M cells. We will integrate these genetic reagents with existing murine disease models to elucidate the role of functional roles of M cells in models of airway inflammation and influenza infection. Lastly, we will dissect the molecular mechanisms of two candidate chemokines that govern M cell-immune cell interactions using genetic and pharmacological manipulation in combination with a novel tracheal explant and lung slice live imaging platform.

M 细胞已被证明可以对管腔抗原进行采样，并使用转胞吞作用将这些抗原传递至底层抗原呈递树突状细胞。因此，它们是粘膜免疫的关键调节因子。尽管身体庞大关于 M 细胞在其他粘膜器官中的免疫作用的文献，气道 M 细胞最近才被描述并且几乎没有进行功能或分子表征。我们提出第一个单细胞转录组小鼠气道 M 细胞并鉴定调节炎症的气道 M 细胞信号级联。我们发现气道 M 细胞在小鼠气管中以孤立细胞的形式出现，在小鼠小气道中以斑块的形式出现，我们报告人气道 M 细胞从原代人气道上皮定向分化。最后，我们证明通过施用信号因子 RANKL、脂多糖 (LPS) 处理来诱导气道 M 细胞，和流感感染。有趣的是，M 细胞可以作为孤立细胞出现，但在 LPS 等生理刺激的情况下或流感感染时，M 细胞斑块与淋巴滤泡相关，表明功能性上皮免疫单元。在本次拨款申请中，我们建议利用我们在干细胞生物学、上皮生物学、肺炎症和流感感染来定义肺 M 细胞的功能生物学。我们将从定义蜂窝开始小鼠和人类气道 M 细胞的起源。我们现在已经生成了一套全面的小鼠遗传驱动程序电池用于追踪气道上皮所有细胞的谱系。在此，我们建议部署这套小鼠系，作为第一个时间，以便精确定义所有可能产生 M 细胞的假定亲代上皮细胞类型。我们也会阐明 RANKL-RANK 信号通路在小鼠炎症模型中的作用。最后，定义气道M细胞功能，我们构建了一个新的 Sox8-CreER 驱动鼠标，它允许我们专门标记、基因修改和消融 M 细胞。我们将把这些遗传试剂与现有的小鼠疾病模型相结合，以阐明 M 细胞在气道炎症和流感感染模型中的功能作用。最后我们来剖析一下分子使用遗传和药理学控制 M 细胞-免疫细胞相互作用的两种候选趋化因子的机制与新型气管外植体和肺切片实时成像平台相结合的操作。