Project 2: CysLT2R Regulation of Type 2 Immunity

摘要/摘要 阿司匹林加重呼吸系统疾病(AERD)的受试者有严重的慢性嗜酸性粒细胞 呼吸道粘膜炎症，半胱氨酰白三烯(Cys- LTS)，肥大细胞(MC)的基础激活，类胰蛋白酶、组胺和 在生物液中检测到前列腺素D2(PGD2)。在当前的资助期，我们报告了 IL-33在AERD患者鼻息肉中高表达。小鼠模型的建立 AERD还显示肺细胞淋巴细胞、IL-33、嗜酸性粒细胞和MC活化增加。 缺乏阿司匹林(ASA)挑战，与西药对照组相比。这些功能中的每一个都是 白三烯C4合成酶(LTC4S)的靶向缺失显著减弱，表明 CysLTs在AERD的2型炎症和基础MC激活中起重要作用 模特。该项目的继续重点是识别CysLT受体(CysLTRs)， CysLTs驱动2型炎症的机制和细胞靶点。中环 假说是CysLT的产生和通过2型cysLTR的信号转导失调 (CysLT2R)在诱发2型炎症和启动MC中起着重要作用。 呼吸道粘膜通过产生IL-33和激活肺组织 内皮细胞。这项提案的发现将对AERD产生直接影响，在 哪些CysLT过度产生，哪些不能完全被1型拮抗剂治疗 CysLTR、CysLT1R。 在目标1中，我们将研究CysLT2R在推动IL-33生成和2型中的作用 使用新的无效菌株的两种小鼠模型中的肺部炎症。我们将验证 CysLT驱动的2型炎症对IL-33的需求，并确定IL-33的细胞来源 33.生产。在目标2中，我们将使用骨髓嵌合体和新的Cysltr2/Flox菌株来 确定调控这些模型的相关CysLT2R表达细胞。在目标3中，我们将使用 从小鼠急性呼吸窘迫综合征模型分离的肺巨噬细胞的转录图谱研究 CysLT2R/IL-33信号通路对肺巨噬细胞合成二十烷类激素的影响 病理生物学功能。我们将使用在几个条件下定义的小鼠MC转录组 为了了解从鼻息肉组织中直接分选的MC的转录图谱 成人急性呼吸窘迫综合征和对照(与项目1合作)。

Summary/Abstract Subjects with aspirin-exacerbated respiratory disease (AERD) have severe chronic eosinophilic inflammation in the respiratory mucosa, excessive production of cysteinyl leukotrienes (cys- LTs), and basal activation of mast cells (MCs) with elevated levels of tryptase, histamine, and prostaglandin D2 (PGD2) detected in biologic fluids. In the current funding period, we reported that IL-33 is highly expressed in the nasal polyps of patients with AERD. The murine model of AERD also demonstrates increased lung cysLTs, IL-33, eosinophils, and MC activation in the absence of aspirin (ASA) challenge, as compared with WT controls. Each of these features is markedly attenuated by the targeted deletion of leukotriene C4 synthase (LTC4S), indicating that cysLTs play a proximal role in driving type 2 inflammation and basal MC activation in the AERD model. The continuation of this project focuses on identifying the cysLT receptors (CysLTRs), mechanisms, and cellular targets by which cysLTs drive type 2 inflammation. The central hypothesis is that dysregulated cysLT production and signaling through the type 2 cysLTR (CysLT2R) play a proximal role in eliciting type 2 inflammation and priming MCs in the respiratory mucosa through the generation of IL-33 and the activation of the pulmonary endothelium. The findings from this proposal will have immediate implications for AERD, in which cysLTs are overproduced and which is incompletely treated by antagonism of the type 1 cysLTR, CysLT1R. In Aim 1, we will examine the role of CysLT2R in driving IL-33 generation and type 2 pulmonary inflammation in two murine models using novel null strains. We will validate the requirement for IL-33 in cysLT-driven type 2 inflammation, and identify the cellular sources of IL- 33 production. In Aim 2, we will use bone marrow chimeras and novel Cysltr2flox/flox strains to identify the relevant CysLT2R-expressing cells that regulate these models. In Aim 3, we will use transcriptional profiling of sorted lung MCs from the murine AERD model to examine the consequences of CysLT2R/IL-33 signaling on priming lung MCs for eicosanoid production and patholobiologic function. We will use the murine MC transcriptome defined in several conditions to understand the transcriptional profile of MCs directly sorted from nasal polyp tissue from adults with AERD and controls (in collaboration with Project 1).