Transcriptional and metabolomic regulation of IL-10 in pulmonary ILC2s

摘要 ILC 2是稳态时肺中占优势的先天淋巴细胞群，它们释放2型 细胞因子是响应嗜酸性粒细胞浸润、增加气道高反应性和相关的呼吸道炎症的中心驱动因子。 肺组织损伤以前，我们的实验室确定了一个ILC 2的子集（ILC 210），它积极地产生和 分泌IL-10，这是一种抗炎细胞因子，具有改善变应性肺炎信号传导的能力（J 过敏临床免疫学，2020年）。重要的是，这些结果已经被其他研究小组在各种研究中证实。 变应性疾病模型（J Exp Med.，2020年，免疫，2021年）。提出研究计划的动机是 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，具有靶向调节的潜力。SA 1旨在探索 调节用于诱导IL-10的转录因子。我们提出了一系列的实验， 慢性过敏性气道炎症模型，以评估关键转录因子cMaf和 Blimp-1首先是通过广泛的、尖端的染色质测序方法，然后是通过逆转录病毒诱导和 敲除小鼠模型。在SA 2中，我们还观察到IL-10的产生显著依赖于关键的代谢产物。 ILC 2经常使用的途径。我们打算通过研究糖酵解、脂肪酸和蛋白质的作用来扩大我们的研究。 酸氧化和信号蛋白AMPK，目的是确定潜在的机制目标， 过敏性疾病的调节疗法。此外，线粒体对IL-10产生的调节与 通过体外和离体线粒体动态测定探索。最后，该项目的两个部分来 共同解决调节致病性ILC 2的特定转录和代谢要求 目的是靶向转化为具有调节气道高反应性能力的ILC 210。结果 从这项研究中获得的信息将为ILC 210在疾病中的重要和未充分研究的作用提供新的见解 与ILC 2相关的疾病，如过敏性肺炎和哮喘。

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.