Role of TNF receptor 2 on Pulmonary Group 2 Innate Lymphoid Cells

摘要 该项目的长期目标是开发更好的呼吸道炎症和过敏疗法。 哮喘。组2先天淋巴样细胞(ILC2)是新近发现的一种产生2型的细胞群 细胞因子对越来越多的环境信号和上皮细胞衍生的细胞因子的反应。 研究表明，在哮喘和许多广泛性疾病中，ILC2活性增加，因为ILC2足以 诱导不依赖适应性免疫的小鼠呼吸道炎症。建议的研究计划是 受我们实验室和其他实验室最近的新观察到的肿瘤坏死因子/TNFR2轴控制ILC2- 依赖的呼吸道炎症(细胞报告2019，J过敏性Clin免疫学。2020)。高水平的肿瘤坏死因子是 发现于哮喘患者的肺部，然而抗肿瘤坏死因子治疗通常与全身性 毒性是由于存在两种截然不同的功能不同的肿瘤坏死因子受体：TNFR1和TNFR2。我们的 结果提示，肿瘤坏死因子可促进ILC2效应细胞因子IL-5和IL-13的分泌 生存通过TNFR2信号，导致呼吸道炎症。然而，肿瘤坏死因子/肿瘤坏死因子受体2轴 对ILC2s的作用机制以及随后的呼吸道炎症的发展仍有待探讨。 根据我们的数据，我们假设阻断ILC2上的TNFR2诱导了一种免疫调节表型 补充不同的代谢来源，共同有利于降低AHR。在具体目标1(SA1)中，我们 我设计了几种方法来表征肿瘤坏死因子对ILC2效应器功能和肺的影响 发炎。我们的初步数据表明，TNFR2在激活的ILC2上异质性表达。我们 因此，我将使用以下组合来表征推动ILC2中肿瘤坏死因子效应的转录因子 单细胞基因组和转录组分析。此外，来自我们实验室和其他实验室的数据表明 ILC2中的代谢过程依赖于脂肪酸氧化产生的能量(FAO)和 氧化磷酸化。有趣的是，我们的结果清楚地显示了TNFR2-/-的代谢向糖酵解的转变 ILC2。基于这些结果，我们打算评估WT和TNFR2-/-ILC2s的糖酵解和FAO机制 在SA2中。最后，我们之前证明了人类ILC2表达TNFR2，而人源化ILC2小鼠 TNFR2依赖的AHR对肿瘤坏死因子的反应。因此，我们打算在SA3中评估我们的 哮喘患者的研究结果。我们将从精心挑选的人群中收集肺和血液中的ILC2 轻/中度、重度哮喘患者和健康献血者，并将BAL中的肿瘤坏死因子水平与 ILC2的数量/表达TNFR2，以及监测其他表达TNFR2的细胞。这些研究， 基于强大的初步数据，将专注于开发过敏性哮喘的新疗法。为了 为了达到这些结果，我们组建了一个团队，其中包括领先的肺部生物学专家和首席 临床肺病学，补充我们在临床前模型方面的丰富经验。

Abstract The long-term goal of this project is to develop better therapies for respiratory inflammation and allergic asthma. Group 2 innate lymphoid cells (ILC2) are a recently identified cell population producing type 2 cytokines in response to a growing number of environmental signals and epithelial cell-derived cytokines. Studies show increased ILC2 activity in asthma and many widespread diseases, as ILC2s are sufficient to induce airway inflammation independent of adaptive immunity in mice. The proposed research plan is motivated by recent new observations from our laboratory and others that the TNF/TNFR2 axis controls ILC2- dependent airway inflammation (Cell Report 2019, J Allergy Clin Immunol. 2020). High levels of TNF are found in the lungs of asthmatic patients, however anti-TNF therapy is generally associated with systemic toxicity due to the existence of two distinct functionally different receptors for TNF: TNFR1 and TNFR2. Our results suggest that TNF enhanced the secretion of ILC2 effector cytokines IL-5 and IL-13 and increased survival via TNFR2 signaling, leading to airway inflammation. However, how the TNF/TNFR2 axis mechanistically affects ILC2s and subsequent development of airway inflammation remains to be explored. Based on our data, we hypothesize that blocking TNFR2 on ILC2s induces an immunoregulatory phenotype fueling on a distinct metabolic source, together favoring the reduction of AHR. In Specific Aim 1 (SA1), we have designed several approaches to characterize the effects of TNF on ILC2 effector functions and lung inflammation. Our preliminary data suggest that TNFR2 is heterogeneously expressed on activated ILC2s. We will therefore characterize the transcription factors driving the effects of TNF in ILC2s using a combination of single cell genomic and transcriptomic analysis. Furthermore, data from our laboratory and others suggest that metabolic processes in ILC2s are dependent on the generation of energy from fatty acid oxidation (FAO) and oxidative phosphorylation. Interestingly our results clearly show a metabolic shift towards glycolysis in TNFR2-/- ILC2s. Based on these results we intend to assess glycolysis and FAO mechanisms in WT and TNFR2-/- ILC2s in SA2. Finally, we previously showed that human ILC2s express TNFR2 and humanized ILC2 mice developed TNFR2-dependent AHR in response to TNF. Therefore, we intend to assess in the SA3 the relevance of our findings in asthmatic patients. We will collect lung and blood ILC2s from carefully selected cohorts of mild/moderate, severe asthmatics and healthy donors and correlate the levels of TNF in the BAL to the numbers of ILC2s/expression of TNFR2, as well as monitor other cells that express TNFR2. These studies, based on strong preliminary data, will focus on developing novel therapy for allergic asthma. In order to achieve these results we have assembled a team including leading experts in lung biology and the chief of clinical pulmonology to complement our extensive experience in pre-clinical models.