Brush cell sensing of aeroallergen-elicited stress signals promotes epithelial cell activation

刷细胞感知空气过敏原引起的应激信号促进上皮细胞活化

• 批准号: 10361506
• 负责人: Lora Bankova
• 金额: $ 21.15万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-02 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10361506
• 关键词: Afferent Neurons; Allergens; Allergic Disease; Alternaria; Apnea; Asthma; Biological Assay; Bronchoconstriction; Brush Cell; Cell Degranulation; Cells; Chronic; Data; Data Set; Eicosanoid Production; Eicosanoids; Enzymes; Epithelial; Epithelial Cells; Event; Foundations; Functional disorder; Gene Expression Profile; Generations; Genetic; Goals; Goblet Cells; Heterogeneity; Human; Immune; Immune response; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Intestinal Mucosa; Intestines; Lead; Ligands; Link; Mediating; Mediator of activation protein; Methods; Mouse Strains; Mucins; Mucous Membrane; Mus; Names; Nasal Epithelium; Nerve; Nose; Olfactory Mucosa; Pathway interactions; Population; Receptor Signaling; Reflex action; Reporting; Respiratory Mucosa; Role; Signal Pathway; Signal Transduction; Site; Sneezing; Source; Specialized Epithelial Cell; Stress; Structure of mucous membrane of nose; System; Taste Buds; Taste Perception; Techniques; Testing; Tissues; Trachea; Vasodilation; Virus; Work; afferent nerve; airborne allergen; allergic airway disease; antimicrobial; autocrine; base; cellular targeting; chronic respiratory disease; chronic rhinosinusitis; cysteinyl leukotriene receptor 2; cysteinyl-leukotriene; cytokine; defined contribution; design; experimental study; in vivo; insight; leukotriene-C4 synthase; lipid mediator; mast cell; neuroinflammation; novel; programs; receptor; respiratory; respiratory reflex; response; sensory system; single-cell RNA sequencing; transcriptome sequencing

PROJECT SUMMARY: Solitary chemosensory cells are rare specialized epithelial cells scattered in the airway (referred to as brush cells) and intestinal mucosa (named tuft cells), recently found to be initiators of type 2 immune responses at least partially through the generation of the proinflammatory cytokine IL-25. In the airways, activation of brush cells by bitter tasting bacterial metabolites also triggers sensory neurons leading to protective airway reflexes. The full effector potential of chemosensory cells and activating receptors beyond taste receptors have not been defined. We have found that epithelial cells sharing the transcriptional profile of chemosensory cells from the trachea and intestine are enriched in the nasal mucosa. We generated a nasal brush cell RNA-seq data set to determine their possible activating receptors and developed an ex vivo system to test the ligand receptor pairs that lead to activation of airway brush cells. We found that brush cells generate large quantities of pro- inflammatory lipid mediators among them cysteinyl leukotrienes (CysLTs). We then generated a new mouse strain with genetic deletion of the terminal CysLT generating enzyme in brush cells to define the contribution of brush cell-derived CysLTs to the pro-inflammatory and protective responses in the airways. In Aim 1, we will define the subsets of nasal brush cells from the respiratory and olfactory mucosa using single cell RNA sequencing. In Aim 2, we will define the brush cell activating pathways triggered in response to aeroallergen sensing, the autocrine loops enhancing this response and the full effector potential of brush cells. In Aim 3, we will define the role of brush cell-derived CysLTs in epithelial cell activation in the airways using mice with genetic deletion of brush cells, CysLTs and brush cell-specific deletion of CysLTs. Findings here will clarify the contribution of brush cell-derived CysLTs to protective and inflammatory responses in the airways and lay the foundation to define their function in human airway mucosa. Results from the proposed experiments will provide critical insights into how protective airway responses designed to expel environmental insults can be diverted to initiate and propagate inflammatory responses leading to allergic airway diseases.

项目概要： 孤立的化学感受细胞是罕见的特化上皮细胞，分散在气道中（称为刷状细胞 细胞）和肠粘膜（称为簇细胞），最近发现是2型免疫应答的启动子， 至少部分通过促炎细胞因子IL-25的产生。在气道中，激活刷 苦味细菌代谢物也触发感觉神经元，导致保护性气道反射。 化学感受细胞和味觉受体以外的激活受体的全部效应潜力尚未被证实。 定义了 我们已经发现，上皮细胞共享来自气管的化学感受细胞的转录谱， 和肠富含于鼻粘膜中。我们生成了鼻刷细胞RNA-seq数据集， 确定它们可能的激活受体，并开发了一种体外系统来测试配体受体对 导致气道刷状细胞的激活。我们发现刷状细胞产生大量的亲- 炎性脂质介质，其中包括半胱氨酰白三烯（CysLTs）。然后我们生成了一只新的老鼠 刷状细胞中末端CysLT生成酶基因缺失的菌株，以确定 刷细胞衍生的CysLT对气道中的促炎和保护性反应的影响。 在目标1中，我们将使用单个细胞的方法来定义来自呼吸道和嗅觉粘膜的鼻刷细胞的亚群。 细胞RNA测序。在目标2中，我们将定义刷状细胞激活途径， 空气过敏原感应，自分泌回路增强这种反应和刷状细胞的全部效应潜力。 在目标3中，我们将使用以下方法定义刷状细胞衍生的CysLT在气道上皮细胞活化中的作用： 具有刷状细胞、CysLT和刷状细胞特异性CysLT缺失的遗传缺失的小鼠。这里的发现将 阐明刷状细胞源性CysLT对气道保护性和炎症反应的作用 为进一步明确它们在人气道黏膜中的功能奠定基础。 从拟议的实验结果将提供关键的见解如何保护气道反应 旨在驱逐环境损害的药物可能会引发和传播炎症反应 导致过敏性气道疾病。