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

• 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
• 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

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.

已经显示M细胞对管腔抗原进行采样，并使用转胞吞作用将这些抗原递送到潜在的细胞。 抗原呈递树突细胞。因此，它们作为粘膜免疫的关键调节剂。尽管有大量的 关于M细胞在其他粘膜器官中的免疫学作用的文献中，气道M细胞只是最近才被描述 并且几乎没有进行功能或分子表征。我们展示了第一个单细胞转录组 的小鼠气道M细胞，并确定气道M细胞信号级联调节炎症。我们已经发现 气道M细胞在小鼠气管中以孤立细胞形式存在，在小鼠小气道中以斑块形式存在，我们报告 从原代人气道上皮细胞定向分化人气道M细胞。最后，我们证明， 通过施用信号传导因子RANKL，用脂多糖（LPS）处理， 和流感感染。有趣的是，M细胞可以作为孤立细胞出现，但在生理刺激如LPS的环境中， 或流感感染时，M细胞斑块与淋巴滤泡相关，提示功能性上皮免疫 单位 在这项拨款申请中，我们建议利用我们在干细胞生物学、上皮生物学、肺生物学、 炎症和流感感染来定义肺M细胞的功能生物学。我们将从定义细胞 小鼠和人气道M细胞的起源。我们现在已经产生了一个全面的电池鼠遗传驱动程序 用于追踪气道上皮的所有细胞的谱系的线。在此，我们建议部署这组鼠系， 时间，以便精确地定义可以产生M细胞的所有假定的亲本上皮细胞类型。我们还将 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Sox 8-CreER驱动小鼠，使我们能够特异性标记，遗传修饰， 消融M细胞。我们将把这些基因试剂与现有的鼠疾病模型相结合，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，我们将剖析分子 两种候选趋化因子利用遗传和药理学方法控制M细胞-免疫细胞相互作用的机制 操作与新型气管外植体和肺切片实时成像平台相结合。