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

基本信息

批准号：
8272435
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
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8272435
关键词：
Abbreviations Acids Acute Affect Agonist American Antigens Antiviral Agents Arginine Asthma Bathing Bronchoconstriction Bronchodilation Calcium Cavia Cells Citrates Contracts Copper Cyclooxygenase Inhibitors Data Disease Dose Electric Stimulation Esters Fluorescein Functional disorder Genes Glutathione Health Histamine Human Imiquimod Immune response In Vitro Indomethacin Inflammatory Lead Left Mediating Modeling Mus Muscle Muscle relaxation phase NG-Nitroarginine Methyl Ester Nature Nerve Neurons Nitric Oxide Nitric Oxide Donors Nitric Oxide Synthase Nitroarginine Pathway interactions Pharmaceutical Preparations Production Prostaglandin Production Prostaglandins RNA Reagent Receptor Signaling Recruitment Activity Relative (related person)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 Uridine Vagus nerve structure Viral Virus Virus Diseases Wheezing Work adapter protein airway hyperresponsiveness airway inflammation antagonist G arginase constriction eosinophil experience human TLR7 protein human TLR8 protein in vivo inhibitor/antagonist large-conductance calcium-activated potassium channels methacholine neuroregulation omega-N-Methylarginine parainfluenza virus paxilline prevent receptor research study respiratory smooth muscle response second messenger sensitizing antigen sildenafil viral RNA

项目摘要

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. PUBLIC HEALTH RELEVANCE: Asthma affects 5 - 10% of Americans. The wheezing and shortness of breath that people with asthma experience is due to constrictions of the airways. We have recently found that drugs working through a receptor that normally recognizes viruses causes the airways to relax. In this project, we will study how these drugs relax the airways, and how this response may be damaged in asthma, to lay the groundwork to allow us to develop this class of drugs into asthma treatments.

描述（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。我们最近发现，刺激 TLR-7 和 TLR-8 可以放松气道平滑肌，并防止支气管因各种刺激而收缩。效果是深远的，可以导致平滑肌几乎完全松弛。我们已经在豚鼠和小鼠以及人类气道组织中证明了这种效应。豚鼠和人类气道中 TLR7 依赖性支气管扩张是由一氧化氮的产生介导的，而 TLR8 依赖性支气管扩张则不是。我们的初步数据表明，TLR8 介导的支气管扩张是通过前列腺素的产生和大电导钙激活钾通道的开放来介导的。我们还表明，抗原致敏和急性病毒感染都会严重损害 TLR 依赖性支气管扩张。在此应用中，我们提出了三个具体目标：具体目标#1。 A) 使用 TLR7(-/-) 小鼠以及新生成的 TLR8(-/-) 和 TLR7(-/-)TLR8(-/-) 小鼠，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影响，B) 在体外测试这些受体是否通过一氧化氮、前列腺素和大电导钙激活钾通道 (BKCa) 发出信号，C) 探索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，一氧化氮和前列腺素的作用以及细胞内钙在这些受体快速信号传导中的变化。具体目标 2：为了测试抗原致敏后 TLR 介导的支气管扩张减少是否是由 TLR7 和/或 TLR8 减少介导，测试目标 #1 中确定的气道平滑肌中的信号通路。由于这些 TLR 位于副交感神经以及招募到气道神经的嗜酸性粒细胞上，因此我们还将测试 TLR7 和 TLR8 是否会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 介导的支气管扩张减少是否是由 TLR7 和/或 TLR8 途径减少介导的，测试目标一中确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对于确定 TLR7 和 TLR8 激动剂治疗哮喘和其他气道疾病的潜力非常重要。此外，由于这些受体对病毒RNA有反应，了解刺激气道中TLR7受体的作用，以及哮喘模型中这些作用的丧失，将有助于我们了解病毒引起的哮喘发作的病理生理学。公共卫生相关性：哮喘影响 5 - 10% 的美国人。哮喘患者出现的喘息和呼吸短促是由于气道收缩造成的。我们最近发现，通过通常识别病毒的受体起作用的药物会导致气道放松。在这个项目中，我们将研究这些药物如何放松气道，以及这种反应如何在哮喘中受到损害，为我们将此类药物开发为哮喘治疗奠定基础。