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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要感觉神经的激活，特别是伤害性 C 纤维，是大多数呼吸系统疾病的一个特征病毒。这种激活的证据可以在 C 纤维激活的经典结果中找到包括打喷嚏、喉咙痛、咳嗽和反射性分泌物。也造成了困扰当出现病毒感染的症状时，这些神经的激活使病毒能够逃离身体并被传播到其他宿主，即伤害感受器激活会放大病毒在社区中的传播。此外，气道迷走神经 C 纤维的激活可导致强烈的反射性支气管收缩和过度的支气管收缩。可能导致哮喘恶化的分泌物，尤其是儿童哮喘。鉴于与人类疾病的相关性，令人惊讶的是，人们对病毒感染如何诱导 C 纤维知之甚少激活和敏化。理论上，病毒感染通过两种常见方式导致 C 纤维激活机制。第一个是上皮细胞的病毒感染导致介质的产生刺激 C 纤维终端。二是病毒本身直接激活神经。第二种机制可能取决于特定的病毒类型。该提案的重点是这第二种（直接）激活机制与冠状病毒有关。我假设冠状病毒刺突蛋白直接与 C 纤维末端相互作用，从而激活和使伤害性 C 纤维敏感。我使用三种正交方法的初步数据支持刺突蛋白直接激活（引起动作电位放电）约 40- 小鼠气道中 50% 的迷走神经 C 纤维。我的第一个目标是描述迷走神经 C 纤维亚型的特征被刺突蛋白激活，并评估刺突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。我的第二个目标集中在机制上。我假设这种相互作用涉及刺突蛋白中半乳糖蛋白的 3 倍，并且独立于刺突蛋白发生受体 ACE2 或 Toll 样受体。无论最近的结合目标如何，我都会解决我们的问题假设激活是继发于 TRPV1 和/或 TRPA1 通道开放的。这些目标将使用气道特异性 mRNA 表达的单细胞 RT-PCR 分析来解决伤害性 C 纤维、细胞外和膜片钳电生理学以及 2 光子实时成像技术。研究结果有望为新的机制提供见解冠状病毒诱导气道 C 纤维激活。