Neuroimmune interactions regulate development of allergic inflammation

项目摘要 哮喘是一种高度流行的气道炎症性疾病，可导致严重的支气管收缩 和呼吸衰竭对过敏原的异常2型免疫应答长期以来一直被认为是 哮喘的主要驱动因素。然而，约10%的患者患有严重的、持续的难以控制的哮喘， 以白细胞介素（IL）-17的产生为标志。这导致嗜肺气道炎症，无论是单独或 与2型炎症结合，2型炎症对目前的治疗（包括皮质类固醇）反应不佳。 这些观察结果强调了鉴定能够调节2型糖尿病和2型糖尿病的分子靶点的重要性。 和17型过敏性肺炎的反应。先天性淋巴样细胞（ILC），最近描述的细胞类型， 已经显示在粘膜组织炎症的引发和扩大中起关键作用。的ilc 2 快速产生促炎细胞因子如IL-5和IL-13，其介导嗜酸性粒细胞募集， 杯状细胞增生，响应于上皮警报素如IL-25和IL-33。另一方面，ILC 3， 产生IL-17，促进和促进嗜肺性哮喘的发展。然而，分子 在ILC 2和ILC 3中促进促炎表型的机制仍然知之甚少， 特别是在过敏性气道炎症的情况下， 我们最近进行了大规模并行的基于液滴的单细胞RNA测序，以开发一种 在稳态和炎症条件下肺驻留ILC的转录图谱，并鉴定新的 ILC功能的调节剂。我们发现两种不同神经肽的受体，神经介肽U（NMU）和 降钙素基因相关肽（CGRP）在肺ILCs上表达。我们的数据显示，NMU直接 在alarmin IL-25的存在下激活ILC以获得促炎表型。而反观 神经肽CGRP在用alarmin刺激后抑制ILC 2产生促炎细胞因子 IL-33。这些数据使我们假设，在过敏原激发后，神经元信号通过神经肽 受体可以有效地调节ILC介导的过敏性炎症。我们提出三个不同的目标， 解决这个假设： 1)通过单细胞转录分析鉴定和验证ILC功能的新调控因子 2）确定NMU/NMUR 1信号传导通过其与ILC 2和ILC 3结合的机制; 将稳态ILC转化为促炎性ILC; 3）研究ILC的作用机制， 神经肽CGRP调节ILC 2功能和变应性肺炎的发展。 总之，这三个目标将创建一个高度整合的先天淋巴细胞（ILC 2）转录图谱 和ILC 3s）在过敏原暴露期间，并阐明神经系统如何调节 通过激活ILC上的神经肽受体引起过敏性气道炎症。 !

PROJECT SUMMARY Asthma is a highly prevalent inflammatory disorder of the airways that can lead to severe bronchoconstriction and respiratory failure. Aberrant type 2 immune responses to allergens have long been appreciated as the major driver of asthma. However, about 10% of patients have severe, persistent difficult to control asthma, marked by the production of interleukin (IL)-17. This leads to neutrophilic airway inflammation, either alone or in conjunction with type 2 inflammation, which responds poorly to current therapies, including corticosteroids. These observations highlight the importance of identifying molecular targets capable of regulating both type 2 and type 17 responses in allergic lung inflammation. Innate lymphoid cells (ILCs), a recently described cell type, have been shown to play a critical role in the initiation and amplification of mucosal tissue inflammation. ILC2s rapidly produce pro-inflammatory cytokines like IL-5 and IL-13, which mediate eosinophil recruitment and goblet cell hyperplasia, in response to epithelial alarmins like IL-25 and IL-33. ILC3s, on the other hand, produce IL-17, and promote and propagate development of neutrophilic asthma. However, the molecular mechanisms that promote pro-inflammatory phenotypes in ILC2s and ILC3s remain poorly understood, particularly in the context of allergic airway inflammation, We recently undertook massively parallel droplet-based single-cell RNA-sequencing to develop a transcriptional atlas of lung resident ILCs under homeostatic and inflammatory conditions and identify novel regulators of ILC function. We found that receptors for two different neuropeptides, neuromedin U (NMU) and calcitonin gene related peptide (CGRP), were expressed on lung ILCs. Our data shows that NMU directly activates ILCs in the presence of the alarmin IL-25 to acquire a pro-inflammatory phenotype. In contrast, the neuropeptide CGRP inhibits ILC2 production of pro-inflammatory cytokines after stimulation with the alarmin IL-33. These data lead us to hypothesize that following allergen challenge, neuronal signals via neuropeptide receptors can potently modulate the ILC-mediated allergic inflammation. We propose three different aims to address this hypothesis: 1) Identify and validate novel regulators of ILC function via single-cell transcriptional analysis of in vivo-derived ILC2s and ILC3s; 2) Determine the mechanism(s) by which NMU/NMUR1 signaling converts homeostatic ILCs into pro-inflammatory ILCs; 3) Study the mechanism by which the neuropeptide CGRP regulates ILC2 function and development of allergic lung inflammation. Together, these three aims will create a highly integrated transcriptional map of innate lymphoid cells (ILC2s and ILC3s) during allergen exposure, and elucidate how the nervous system may regulate development of allergic airway inflammation by activating neuropeptide receptors on ILCs. !