ACUTE AIRWAY EFFECTS OF TLR7 AND TLR8 STIMULATION IN HEALTH AND DISEASE

TLR7 和 TLR8 刺激对健康和疾病的急性气道影响

• 批准号: 9014554
• 负责人: David B Jacoby
• 金额: $ 43.54万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9014554
• 关键词: Acute; Affect; Agonist; American; Antigens; Antiviral Agents; Asthma; Bronchodilation; Calcium; Cavia; Cells; Data; Disease; Functional disorder; Genes; Health; Human; Immune response; In Vitro; Inflammatory; Lead; Mediating; Modeling; Mus; Muscle relaxation phase; Nerve; Neurons; Nitric Oxide; Pathway interactions; Pharmaceutical Preparations; Production; Prostaglandin Production; Prostaglandins; RNA; Receptor Signaling; Recruitment Activity; Relaxation; Role; Second Messenger Systems; Shortness of Breath; Signal Pathway; Smooth Muscle; Smooth Muscle Myocytes; Stimulus; TLR7 gene; TLR8 gene; Testing; Tissues; Toll-like receptors; Viral; Virus; Virus Diseases; Wheezing; Work; adapter protein; airway hyperresponsiveness; airway inflammation; constriction; eosinophil; experience; large-conductance calcium-activated potassium channels; neuroregulation; parainfluenza virus; prevent; receptor; research study; respiratory smooth muscle; response; second messenger; sensitizing antigen; viral RNA

DESCRIPTION (provided by applicant): Viral single standed RNA is sensed by cells via toll-like receptor (TLR)-7 and TLR-8. We have recently shown that stimulating TLR-7 and TLR-8 relaxes airway smooth muscle and prevents broncho-constriction in response to a variety of stimuli. The effect is profound, and can lead to virtually complete relaxation of smooth muscle. We have demonstrated this effect in guinea pigs and mice, and in human airway tissues. TLR7 dependent broncho-dilation in guinea pigs and in human airways is mediated by production of nitric oxide, while TLR8 dependent broncho-dilation is not. Our preliminary data suggest that TLR8 mediated broncho-dilation is mediated by production of prostaglandins and opening of the large conductance, calcium activated potassium channel. We have also shown that both antigen sensitization and acute viral infection substantially impair TLR- dependent broncho-dilation. In this application, we propose three specific aims: Specific Aim #1. To A) use TLR7(-/-) mice, as well as newly generated TLR8(-/-) and TLR7(-/-)TLR8(-/-) mice, to more thoroughly investigate and establish the signaling pathways responsible for the effects of these receptors on airway neurons and smooth muscle, B) test whether these receptors signal through nitric oxide, prostaglandins, and the large conductance, calcium activated potassium channel (BKCa) in human airways in vitro, and C) explore the role of nitric oxide and prostaglandins and changes in intracellular calcium in rapid signaling by these receptors in primary cultures of human airway parasympathetic neurons and airway smooth muscle cells. Specific Aim 2: To test whether decreased TLR mediated broncho-dilation after antigen sensitization is mediated by decreased TLR7 and/or TLR8, testing signaling pathways in airway smooth muscle identified in Aim #1. Since these TLRs are on parasympathetic nerves, as well as on eosinophils recruited to the airway nerves, we will also test whether TLR7 and TLR8 change neural control in sensitized airways. SPECIFIC AIM #3: To test whether decreased TLR mediated broncho-dilation after viral infection is mediated by decreased TLR7 and/or TLR8 pathways, testing all second messenger signaling pathways in airway smooth muscle identified in aim one. We will also investigate the role of changes in parasympathetic function. The results of the experiments we propose will be important in establishing the potential of TLR7 AND TLR8 agonists as treatments for asthma and other airway diseases. In addition, because these receptors respond to viral RNA, understanding the effects of stimulating TLR7 receptors in the airways, as well as the loss of these effects in models of asthma, will help us understand the pathophysiology of virus induced asthma attacks.

描述（由申请人提供）：病毒单链RNA通过toll样受体（TLR）-7和TLR-8被细胞感知。我们最近已经表明，刺激TLR-7和TLR-8松弛气道平滑肌，并防止支气管收缩，以响应各种刺激。效果是深远的，并可导致平滑肌几乎完全放松。我们已经在豚鼠和小鼠以及人体气道组织中证明了这种效应。豚鼠和人气道中的TLR 7依赖性支气管扩张是由一氧化氮的产生介导的，而TLR 8依赖性支气管扩张不是。我们的初步数据表明，TLR 8介导的支气管扩张是通过产生洋地黄素和打开大电导，钙激活钾通道介导的。我们还表明，抗原致敏和急性病毒感染实质上损害TLR依赖性支气管扩张。在本申请中，我们提出了三个具体目标：具体目标#1。A）使用TLR 7（-/-）小鼠，以及新产生的TLR 8（-/-）和TLR 7（-/-）TLR 8（-/-）小鼠，以更彻底地研究和建立负责这些受体对气道神经元和平滑肌的作用的信号传导通路，B）测试这些受体是否通过一氧化氮、洋地黄素和大电导进行信号传导，体外人气道中钙激活钾通道（BKCa），和C）在人气道副交感神经元和气道平滑肌细胞的原代培养物中，探索一氧化氮和洋地黄素的作用以及细胞内钙在这些受体的快速信号传导中的变化。具体目标二：为了测试抗原致敏后TLR介导的支气管扩张的减少是否由TLR 7和/或TLR 8的减少介导，测试目的#1中确定的气道平滑肌中的信号传导途径。由于这些TLR在副交感神经上，以及在募集到气道神经的嗜酸性粒细胞上，我们还将测试TLR 7和TLR 8是否改变致敏气道中的神经控制。具体目标3：为了测试病毒感染后TLR介导的支气管扩张的降低是否由降低的TLR 7和/或TLR 8途径介导，测试目的一中鉴定的气道平滑肌中的所有第二信使信号传导途径。我们还将研究副交感神经功能变化的作用。我们提出的实验结果将是重要的，在建立TLR 7和TLR 8激动剂作为治疗哮喘和其他气道疾病的潜力。此外，由于这些受体对病毒RNA有反应，因此了解刺激气道中TLR 7受体的作用以及这些作用在哮喘模型中的丧失将有助于我们了解病毒诱导的哮喘发作的病理生理学。