Vagal airway sensory nerve activation by beta-coronavirus spike protein

β-冠状病毒刺突蛋白激活迷走神经气道感觉神经

• 批准号: 10748485
• 负责人: Joyce Sooyeon Kim
• 金额: $ 6.95万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10748485
• 关键词: 2019-nCoV; ACE2; Action Potentials; Address; Affinity; Alveolus; Angiotensin Receptor; Asthma; Binding; Bronchi; Bronchoconstriction; C Fiber; COVID-19 pandemic; Cell membrane; Cells; Child; Chronic; Chronic Obstructive Pulmonary Disease; Common Cold; Communities; Coronavirus; Coronavirus spike protein; Coughing; Data; Disease; Electrophysiology (science); Embryo; Epithelial Cells; Fellowship; Functional disorder; Goals; Imaging Techniques; Individual; Infection; Ion Channel; Lead; Lung; Mediating; Mediator; Membrane; Membrane Glycoproteins; Middle East Respiratory Syndrome Coronavirus; Molecular; Mus; Nerve; Nervous System; Neural Crest; Neurons; Nociception; Nociceptors; Nodose Ganglion; Nose; Oropharyngeal; Phenotype; Process; Production; Protein Subunits; Proteins; Reflex action; Research Proposals; Respiratory Disease; Respiratory Mucosa; Respiratory System; Respiratory Tract Infections; Reverse Transcriptase Polymerase Chain Reaction; SARS coronavirus; SARS-CoV-2 spike protein; Secondary to; Sensory; Sneezing; Sore Throat; Spinal; Spinal Ganglia; Stimulus; Surface; Symptoms; TLR4 gene; TRPA channel; TRPV1 gene; Toll-like receptors; Trachea; Vertebral column; Viral; Virus; Virus Diseases; activated Protein C; afferent nerve; asthma exacerbation; betacoronavirus; extracellular; human disease; insight; mRNA Expression; novel; patch clamp; receptor; respiratory; respiratory virus; theories; transmission process; two-photon; virtual; virus morphology

PROJECT SUMMARY Activation of sensory nerves, in particular nociceptive C-fibers, is a feature of most respiratory viruses. Evidence of such activation is found in the classical consequences of C-fiber activation including sneezing, sore throat, coughing, and reflex secretions. As well as causing the troubling symptoms of viral infection, the activation of these nerves allows viruses to escape the body and be transmitted to other hosts, i.e. nociceptor activation amplifies viral spread in a community. In addition, activation of airway vagal C-fibers can lead to strong reflex bronchoconstriction and excessive secretions that likely contribute to the exacerbation of asthma particularly in children. Given the relevance to human disease, surprisingly little is known about how virus infection induces C-fiber activation and sensitization. In theory, viral infection leads to C-fiber activation by two general mechanisms. The first is that viral infection of epithelial cells leads to the production of a mediator(s) that stimulates the C-fiber terminals. The second is that the virus itself directly activates the nerves. This second mechanism will likely be dependent on the specific virus type. This proposal focuses on this second (direct) mechanism of activation as it relates to coronaviruses. I hypothesize that the coronavirus spike protein interacts directly with C-fiber terminals in a manner that activates and sensitizes the nociceptive C-fibers. My preliminary data, using three orthogonal approaches, support the conclusion that the spike protein directly activates (evokes action potential discharge) about 40- 50% of vagal C-fibers in mouse airways. My first aim is to characterize the subtype of vagal C-fibers that are activated by spike protein and also to assess whether the spike protein, short of overt activation, leads to the sensitization of C-fiber terminals, i.e. renders them more sensitive to other activating stimuli. My second aim focuses on the mechanism. I hypothesize that this interaction involves the galactin-3 fold in the spike protein, and occurs independently of the spike protein receptor ACE2 or toll-like receptors. Irrespective of the proximal binding target, I will address our hypothesis that activation is secondary to the opening of TRPV1 and or TRPA1 channels. These aims will be addressed using single cell RT-PCR analysis of mRNA expression in airway specific nociceptive C-fibers, extracellular and patch-clamp electrophysiology, and 2-photon live imaging techniques. The results of the studies are expected to provide insights into a novel mechanism of coronavirus induced airway C-fiber activation.

项目摘要 感官神经的激活，尤其是伤害感受性C纤维，是大多数呼吸系统的特征 病毒。在C纤维激活的经典后果中发现了这种激活的证据 包括打喷嚏，喉咙痛，咳嗽和反射分泌物。以及引起令人不安的 病毒感染的症状，这些神经的激活使病毒能够逃脱身体并成为 传输到其他宿主，即伤害感受器激活在社区中扩大病毒传播。此外， 气道迷走神经C纤维的激活会导致强烈的反射支气管收缩和过量 可能导致哮喘加剧的分泌物，特别是在儿童中。鉴于 与人类疾病有关，令人惊讶的是，关于病毒感染如何诱导C纤维知之甚少 激活和敏化。从理论上讲，病毒感染导致两个一般的C纤维激活 机制。首先是上皮细胞的病毒感染导致产生介体 刺激C纤维端子。第二个病毒本身直接激活神经。 该第二种机制可能取决于特定的病毒类型。该提议重点 与冠状病毒相关的第二个（直接）激活机理。我假设 冠状病毒尖峰蛋白直接与C纤维末端相互作用，以激活和 使伤害性C纤维敏感。我的初步数据，使用三种正交方法，支持 尖峰蛋白直接激活（引起动作电位放电）的结论约为40-- 小鼠气道中50％的迷走神经纤维。我的第一个目的是描述迷走神经的子类型 被峰值蛋白激活，还评估峰值蛋白是否缺乏明显的 激活，导致C纤维终端的敏化，即使它们对其他 激活刺激。我的第二个目标重点是机制。我假设这种互动 涉及峰值蛋白中的Galactin-3倍，并独立于峰值蛋白 受体ACE2或Toll样受体。无论近端约束目标，我都会解决我们的 假设激活是TRPV1和或TRPA1通道的开放次要的。这些目标 将使用气道特异性中mRNA表达的单细胞RT-PCR分析来解决 伤害性C纤维，细胞外和贴片钳电生理学以及2光子活成像 技术。预计研究结果将提供有关一种新型机制的见解 冠状病毒诱导气道C纤维激活。