Transcriptional and metabolomic regulation of IL-10 in pulmonary ILC2s

摘要 ILC2是肺中主要的天然淋巴样细胞群，处于稳定状态，它们释放2型 细胞因子是反应嗜酸性粒细胞浸润、增加呼吸道高反应性和与之相关的中心驱动因素 肺组织损伤。此前，我们的实验室发现了ILC2s(ILC210)的一个子集，它活跃地产生和 分泌IL-10，一种抗炎细胞因子，具有改善过敏性肺部炎症信号的能力(J 过敏性Clin免疫学，2020)。重要的是，这些结果已经被其他组织在各种不同的 过敏性疾病模型(J Exp Med.，2020，免疫力，2021)。拟议的研究计划的动机是 最近的初步观察表明，关键的分子和转录要求可能是 诱导IL-10所需的，具有靶向调节的潜力。SA1旨在探索 转录因子对IL-10诱导的调节作用。我们提出了一系列关于急性和慢性疾病的实验 慢性过敏性气道炎症模型评估关键转录因子cMaf和cMaf的参与 BLIMP-1首先通过扩张性的、尖端的染色质测序方法，然后通过逆转录病毒诱导和 敲除小鼠模型。在SA2中，我们还观察到IL-10的产生在很大程度上依赖于关键代谢 ILC2经常使用的途径。我们打算通过研究糖酵解的作用来扩大我们的研究范围，脂肪 酸性氧化和信号蛋白AMPK，目的是确定潜在的机制靶点 过敏性疾病的调制疗法。此外，BE对线粒体产生IL-10的调节作用 通过体外和体外线粒体动态分析进行了探索。最后，这个项目的两个部分来了 共同解决调节致病ILC2的特定转录和代谢需求 目的是有针对性地转换为ILC210，具有调节呼吸道高反应性的能力。结果是 这项研究将为ILC210在疾病中的重要和未被研究的作用提供新的见解 与ILC2相关的疾病，如过敏性肺部炎症和哮喘。

Abstract ILC2s are the dominant innate lymphoid cell population in the lungs at steady state and their release of type-2 cytokines is a central driver in responding eosinophil infiltration, increased airway hyperreactivity and associated lung tissue injury. Previously, our laboratory identified a subset of ILC2s (ILC210s) that actively produce and secrete IL-10, an anti-inflammatory cytokine with the ability to ameliorate allergic lung inflammation signaling (J Allergy Clin Immunol., 2020). Importantly, these results have been confirmed by other groups in a variety of allergic disease models (J Exp Med., 2020, Immunity, 2021). The proposed research plan is motivated by recent preliminary observations demonstrating that key molecular and transcriptional requirements may be required for the induction of IL-10, with the potential for targeted modulation. SA1 is intended to explore the regulation of transcription factors for the induction of IL-10. We propose a series of experiments in acute and chronic models of allergic airway inflammation to assess the involvement of key transcription factors cMaf and Blimp-1 first by expansive, cutting-edge chromatin sequencing methods, and next by retroviral induction and knock-out mouse models. In SA2, we also observed that production of IL-10 relies significantly on key metabolic pathways often utilized by ILC2s. We intend to expand our studies by investigating the role of glycolysis, fatty acid oxidation and signaling protein AMPK, with the aim of identifying mechanistic targets for the potential modulatory therapies for allergic disease. Additionally, mitochondrial regulation of IL-10 production with be explored through in vitro and ex vivo mitochondrial dynamic assays. Finally, the two parts of this project come together to address specific transcriptional and metabolic requirements for the modulation of pathogenic ILC2s with the intention of targeted conversion to ILC210s with the ability to regulate airway hyperreactivity. The results obtained from this study will provide novel insights into an important and understudied role of ILC210s in diseases associated with ILC2s such as allergic lung inflammation and asthma.