Beta-2 adrenergic signaling in immune homeostasis and reconstitution

项目摘要 神经递质和荷尔蒙是连接神经和免疫系统的纽带。其中， 去甲肾上腺素和肾上腺素是在交感神经节后神经元合成的 系统和肾上腺髓质。释放后，它们与β2-肾上腺素能受体(β2AR)结合，表达于 免疫细胞。值得注意的是，β2AR的异常表达和基因多态与几种类型相关 自身免疫性疾病。然而，β2AR信号功能障碍的机制导致了这些 疾病在很大程度上仍不为人所知。在这种情况下，我们对小鼠模型的研究已经揭示了 未被认识的β2AR在免疫发展和重建中的主要二分性作用：1)在特定的 在无病原体条件下，β2AR缺陷导致胸腺T细胞发育显著减少 对比骨髓(BM)中CD115+髓系细胞发育的增加；2)胸腺和髓系 在同种异体移植后的免疫重建中，发育表型明显更明显 3)β-2AR在成熟外周T细胞中的不同作用 在移植物抗宿主病(GVHD)中导致不同结局的亚群和髓系细胞亚群。一起， 这些发现突显了β2AR信号在维持免疫稳态中的关键和基础作用 调节免疫反应。因此，这项研究将追求三个具体目标，以测试一个 β2AR信号调节免疫发育和重建的假说 规范的免疫信号通路和/或微调免疫细胞的新陈代谢适应性。目标一号将决定 β-2AR信号调节T细胞发育、重建和移植物抗宿主病的机制目标2将定义 β-2AR信号调节CD115+髓系细胞发育、重建和移植物抗宿主病的机制 目的3研究药理学β-2AR干预的翻译潜力和机制。我们将使用 RNA-Seq无偏转录技术与流式细胞术、细胞飞行时间质谱仪和海马 代谢试验结合细胞类型特异性β2AR基因敲除(Cre-loxP)以确定分子机制 用于T细胞和髓系细胞的分化和功能。在allo-HCT的背景下，我们还将审查其影响 这些机制对移植物抗宿主病和移植物抗宿主病的影响。这个项目不仅将提高我们对 中枢和外周获得性免疫细胞和天然免疫细胞中β-2AR信号转导的基本机制 免疫器官，但这也可能解释β2AR信号如何在免疫疾病和异基因红细胞移植中起作用 基础免疫疗法。因此，这项研究可能会对治疗的理论基础产生重要的影响 在操纵β2AR信号时。由于β2AR激动剂和拮抗剂已经广泛使用，新的 这项研究产生的临床试验可以迅速应用于接受allo-hct治疗的患者 治疗血癌或其他血液或免疫学疾病，在这些疾病中，allo-hct可以治愈。

Project Summary Neurotransmitters and hormones serve as a link between the nervous and immune systems. Among them, norepinephrine and epinephrine are synthesized in the postganglionic neurons of the sympathetic nervous system and the adrenal medulla. Upon release, they engage the β2-adrenergic receptor (β2AR) expressed on immune cells. Notably, β2AR abnormal expression and gene polymorphisms are associated with several types of autoimmune diseases. However, mechanisms by which β2AR signaling dysfunction contribute to these diseases remain largely unknown. In this context, our studies with mouse models have uncovered previously unappreciated major dichotomous roles of β2AR in immune development and reconstitution: 1) Under specific pathogen free (SPF) condition, β2AR deficiency causes significantly reduced T cell development in thymus in contrast to increased CD115+ myeloid cell development in bone marrow (BM); 2) Both thymic and myeloid development phenotypes are remarkably more manifested during immune reconstitution following allogeneic hematopoietic cell transplantation (allo-HCT); 3) β2AR plays differential roles in in mature peripheral T cell subsets and myeloid cell subsets that result in different outcome in graft-versus-host disease (GVHD). Together, these findings highlight the critical and fundamental role of β2AR signaling in maintaining immune homeostasis and regulating immune response. Therefore, this study will pursue three specific aims to test an overarching hypothesis that β2AR signaling regulates immune development and reconstitution via cross-talking with canonical immune signaling pathways and/or fine-tuning metabolic fitness of immune cells. Aim 1 will determine mechanisms by which β2AR signaling regulates T cell development, reconstitution and GVHD. Aim 2 will define mechanisms by which β2AR signaling regulates CD115+ myeloid cell development, reconstitution and GVHD. Aim 3 will study the translational potential and mechanisms of pharmacologic β2AR intervention. We will use unbiased RNA-seq transcriptomic approach along with flow cytometry, CyTOF mass cytometry and Seahorse metabolic assay combined with cell type-specific β2AR knockout (Cre-LoxP) to define the molecular mechanisms for T cell and myeloid cell differentiation and function. In the setting of allo-HCT, we will also examine the impact of these mechanisms on GVHD and GVT effect. This project will not only improve our understanding of the fundamental mechanisms of β2AR signaling in adaptive and innate immune cells in central and peripheral immune organs, but it may also explain how β2AR signaling contributes to immunologic disorders and allo-HCT based immunotherapy. Therefore, this study may have important ramifications on therapeutic rationale based upon manipulation of β2AR signaling. Since β2AR agonists and antagonists are already widely available, new clinical trials emanating from this research could be rapidly implemented for patients receiving allo-HCT for treatment of blood cancers, or other hematologic or immunologic diseases in which allo-HCT can be curative.