Transcriptional and metabolomic regulation of IL-10 in pulmonary ILC2s

摘要 ILC2 是稳态下肺部主要的先天淋巴细胞群，它们释放 2 型 细胞因子是响应嗜酸性粒细胞浸润、气道高反应性增加和相关 肺组织损伤。此前，我们的实验室鉴定了 ILC2 的一个子集 (ILC210)，它们主动产生和 分泌 IL-10，一种抗炎细胞因子，能够改善过敏性肺部炎症信号传导 (J 过敏临床免疫学，2020）。重要的是，这些结果已被多个领域的其他小组证实。 过敏性疾病模型（J Exp Med.，2020，Immunity，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，具有靶向调节的潜力。 SA1 旨在探索 诱导 IL-10 的转录因子的调节。我们提出了一系列急性和 过敏性气道炎症的慢性模型，以评估关键转录因子 cMaf 和 Blimp-1 首先通过广泛的、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 的产生显着依赖于关键代谢 ILC2 经常利用的途径。我们打算通过调查糖酵解、脂肪的作用来扩大我们的研究 酸氧化和信号蛋白 AMPK，旨在识别潜在的机制靶标 过敏性疾病的调节疗法。此外，线粒体对 IL-10 产生的调节 通过体外和离体线粒体动态测定进行探索。最后，这个项目的两个部分来了 共同解决致病性 ILC2 调节的特定转录和代谢要求 目的是有针对性地转换为能够调节气道高反应性的 ILC210。结果 从这项研究中获得的结果将为 ILC210 在疾病中的重要且尚未得到研究的作用提供新的见解 与 ILC2 相关，例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.