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）：病毒单架RNA通过Toll样受体（TLR）-7和TLR-8感测。我们最近表明，刺激TLR-7和TLR-8放松气道平滑肌，并防止对各种刺激的响应支气管收缩。效果是深刻的，可以导致平滑肌的完全放松。我们已经在豚鼠和小鼠以及人类气道组织中证明了这种作用。豚鼠和人气中的TLR7依赖性支气管稀释是通过一氧化氮的产生介导的，而TLR8依赖的支气管脱丝不是。我们的初步数据表明，TLR8介导的支气管稀释是由前列腺素的产生和大电导的开放，即钙活化钾通道介导的。我们还表明，抗原敏化和急性病毒感染显着损害了TLR依赖性的支气管降解。在此应用程序中，我们提出了三个特定目标：特定目标＃1。 a）使用TLR7（ - / - ）小鼠，以及新产生的TLR8（ - / - ）和TLR7（ - / - ）TLR8（ - / - ）小鼠，更彻底地调查并建立负责这些受体对这些受体对气道神经元和平滑肌的影响的效果的信号传导途径，b）是否通过NITROCTIMS和平滑的氧化物来测试，B）钾通道（BKCA）体外人类气道中的钾通道，以及c）探索一氧化氮和前列腺素的作用，以及这些受体在人类气道副交感神经元和气道平滑肌细胞中这些受体在快速信号传导中的作用。具体目标2：测试抗原致敏后TLR介导的支气管降低是否通过降低的TLR7和/或TLR8介导，在AIM＃1中鉴定出的气道平滑肌中测试信号通路。由于这些TLR处于副交感神经以及被招募到气道神经的嗜酸性神经上，我们还将测试TLR7和TLR8是否会改变敏化气道中的神经控制。特定目的＃3：测试病毒感染后TLR介导的支气管降低是否通过降低的TLR7和/或TLR8途径介导，测试AIM ONE中鉴定出的气道平滑肌中的所有第二个Messenger信号通路。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对于确定TLR7和TLR8激动剂作为哮喘和其他气道疾病的治疗的潜力很重要。此外，由于这些受体对病毒RNA有反应，因此了解气道中刺激的TLR7受体的作用，以及在哮喘模型中这些作用的丧失，将有助于我们了解病毒诱导的哮喘攻击的病理生理学。