The role of airway epithelial CRAC channels in allergic lung inflammation

气道上皮CRAC通道在过敏性肺部炎症中的作用

• 批准号: 9911517
• 负责人: Timothy Kountz
• 金额: $ 4.07万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9911517
• 关键词: ATP Receptors; Air; Airway Disease; Allergens; Allergic; Allergic inflammation; Asthma; Automobile Driving; Binding; Biological Assay; Biological Phenomena; Biological Process; Bronchoalveolar Lavage Fluid; Bronchodilation; Calcium; Calcium Channel; Cause of Death; Cell physiology; Cells; Chronic; Chronic Obstructive Airway Disease; Cytosol; Data; Dinoprostone; Disease; Disease model; Enzyme-Linked Immunosorbent Assay; Enzymes; Epithelial Cells; Estrogen receptor positive; Exocytosis; Extracellular Space; Genetic Transcription; Goblet Cells; Histology; Image; Immune; Immune response; In Vitro; Inflammation; Inflammatory; Ion Channel; Knock-out; Knockout Mice; Lead; Ligands; Link; Lipids; Liquid substance; Luciferases; Lung; Lung Inflammation; Lung diseases; Maintenance; Measures; Mediating; Mentorship; Modeling; Molecular; Molecular Biology; Mucins; Mucous body substance; Mus; Nucleotides; PTGS2 gene; Pathway interactions; Pattern; Pattern recognition receptor; Phospholipase A2; Phosphorylation; Plug-in; Process; Production; Prostaglandins; Proteins; Pyroglyphidae; Receptor Signaling; Regulation; Research; Respiratory physiology; Role; Route; STIM1 gene; Second Messenger Systems; Severe Combined Immunodeficiency; Signal Transduction; T-Lymphocyte; Testing; Therapeutic; Tissues; Total Internal Reflection Fluorescent; Vasodilation; Vesicle; Western Blotting; airway epithelium; airway hyperresponsiveness; airway inflammation; asthma model; asthmatic; asthmatic patient; cyclooxygenase 1; cytokine; enzyme activity; extracellular; histological stains; immunoregulation; in vivo; live cell imaging; loss of function mutation; mouse model; mucin hypersecretion; new therapeutic target; novel; prevent; receptor; recruit; response; transcription factor

Abstract: For many years, the primary responsibility of airway epithelial cells was considered to be the maintenance of barrier function. Recently, airway epithelial cells (AECs) have emerged as key orchestrators of immune responses. For instance, AECs can respond to allergen presence in the airway lumen and thereby direct immune cells, promoting allergic sensitization. The allergens can activate pattern recognition receptors on AECs leading to the production of immunostimulatory lipids and proteins. Many of these receptors transduce signals through the ubiquitous signaling ion Ca2+. Another class of molecules that can activate receptors on AECs is damage-associated molecular patterns (DAMPs). One DAMP that has been linked to Ca2+ signaling and asthma is ATP. Extracellular ATP is known to contribute both to allergic sensitization and chronic inflammation. These effects are likely linked to ATP-mediated production of cytokines, prostaglandins and mucin proteins from AECs. We recently discovered Calcium Release-Activated Calcium (CRAC) channels are a major mechanism for Ca2+ signaling in AECs ultimately driving cytokine production. Preliminary data suggests that ATP receptors can activate CRAC channels in AECs. However, how CRAC channels interface with ATP signaling to drive inflammatory processes is unclear. Therefore, in this proposal, we will further interrogate the significance of AEC CRAC channels in airway inflammation and disease models of asthma. We will do this through the following aims. Aim 1: Determine the mechanism for how AEC CRAC channel activation induces prostaglandin E2 synthesis. Aim 2: Investigate the role of AEC CRAC channels in receptor-stimulated exocytosis. Aim 3: Define the role of AEC CRAC channels in allergic lung inflammation in vivo. We will approach these studies through the use of an integrated analysis using calcium imaging, molecular biology, ELISAs, live cell imaging, tissue-specific knockout mice, histology and assessment of lung function. This research will occur under the direct mentorship of Dr. Murali Prakriya. Collectively, this research has the potential to unveil new targets to inhibit the chronic inflammation driving asthma.

摘要： 多年来，气道上皮细胞的主要责任被认为是呼吸道上皮细胞的功能。 保持屏障功能。近年来，气道上皮细胞（AECs）已成为 免疫反应的协调。例如，AEC可以对皮肤中存在的过敏原做出反应。 气道腔，从而直接免疫细胞，促进过敏致敏。过敏原可以 激活AEC上的模式识别受体，导致产生免疫刺激性脂质 和蛋白质。这些受体中的许多通过普遍存在的信号传导离子Ca 2+传递信号。 另一类可以激活AEC上受体的分子是损伤相关分子， 模式（DAMP）。一种与Ca2+信号传导和哮喘有关的DAMP是ATP。 已知细胞外ATP有助于过敏性致敏和慢性炎症。这些 这种作用可能与ATP介导的细胞因子、胰高血糖素和粘蛋白的产生有关 从AECs我们最近发现钙释放激活钙（CRAC）通道是一种 这是AEC中Ca2+信号传导最终驱动细胞因子产生的主要机制。初步数据 表明ATP受体可以激活AEC中的CRAC通道。然而，CRAC通道如何 与ATP信号传导的接口驱动炎症过程尚不清楚。因此，在本提案中， 我们将进一步研究AEC CRAC通道在气道炎症中的意义， 哮喘的疾病模型我们将通过以下目标来实现这一目标。目标1：确定 AEC CRAC通道激活如何诱导前列腺素E2合成的机制。目标二： 研究AEC CRAC通道在受体刺激的胞吐作用中的作用。目标3：定义角色 AEC CRAC通道在体内变应性肺炎中的作用。我们将通过以下方式进行这些研究： 使用钙成像、分子生物学、ELISA、活细胞成像等综合分析， 组织特异性敲除小鼠，组织学和肺功能评估。这项研究将发生 在穆拉利·普拉卡什博士的直接指导下总的来说，这项研究有可能 揭示了新的目标，以抑制慢性炎症驱动哮喘。