Neuroimmune interactions regulate development of allergic inflammation

项目总结 哮喘是一种高度流行的呼吸道炎症性疾病，可导致严重的支气管收缩。 和呼吸衰竭。对过敏原的异常2型免疫反应长期以来被认为是 哮喘的主要驱动力。然而，约10%的患者患有严重的、持续性的难以控制的哮喘， 以白介素17的产生为特征。这会导致中性粒细胞呼吸道炎症，无论是单独的还是 与2型炎症相结合，后者对目前的治疗方法反应不佳，包括皮质类固醇。 这些观察结果突显了识别能够调节两种类型2的分子靶点的重要性 变态反应性肺部炎症的17型反应。先天淋巴样细胞(ILCs)，一种最近被描述的细胞类型， 已被证明在粘膜组织炎症的启动和扩大中起关键作用。ILC2s 迅速产生促炎细胞因子，如IL-5和IL-13，它们介导嗜酸性粒细胞募集和 杯状细胞增生，对IL-25和IL-33等上皮性警报的反应。另一方面，ILC3， 产生IL-17，促进和促进中性粒细胞哮喘的发展。然而，分子 ILC2和ILC3促进促炎表型的机制仍然知之甚少。 尤其是在过敏性呼吸道炎症的情况下， 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 肺内ILC在动态平衡和炎症状态下的转录图谱和鉴定新的 ILC功能的调节器。我们发现两种不同神经肽的受体，神经梅毒素U(NMU)和 肺ILCs表达降钙素基因相关肽(CGRP)。我们的数据显示NMU直接 在警报蛋白IL-25存在的情况下激活ILCs，以获得促炎表型。相比之下， 神经肽CGRP抑制警报刺激后ILC2促炎细胞因子的产生 IL-33。这些数据使我们假设，在过敏原攻击之后，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。我们提出了三个不同的目标来 回答这个假设： 1)通过对IN的单细胞转录分析，识别和验证ILC功能的新调节因子 活体来源的ILC2和ILC3；2)决定NMU/NMUR1信号转导的机制(S 将体内稳态的ILCs转化为促炎ILCs；3)研究 神经肽CGRP调节ILC2功能和过敏性肺炎的发生发展。 这三个目标结合在一起，将创建一个高度整合的先天淋巴样细胞(ILC2)转录图谱 和ILC3s)，并阐明神经系统如何调节 通过激活ILCs上的神经肽受体引起过敏性呼吸道炎症。 好了！

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. !