Project 2: CysLT2R Regulation of Type 2 Immunity

摘要/摘要 患有阿司匹林加重的呼吸道疾病（AERD）的受试者患有重度慢性嗜酸性粒细胞增多症， 呼吸道粘膜炎症、半胱氨酰白三烯（cys- LT），肥大细胞（MC）的基础活化，类胰蛋白酶、组胺和 前列腺素D2（PGD 2）在生物液体中检测。在当前的融资期内，我们报告了 IL-33在AERD患者的鼻息肉中高度表达。的鼠模型 AERD还表明，肺内cysLT、IL-33、嗜酸性粒细胞和MC活化增加， 与WT对照相比，不存在阿司匹林（阿萨）激发。这些特征中的每一个都 通过靶向缺失白三烯C4合酶（LTC 4S）而显著减弱，表明 cysLT在驱动AERD中的2型炎症和基础MC活化中起近端作用 模型该项目的继续重点是鉴定cysLT受体（CysLTRs）， 机制和cysLT驱动2型炎症的细胞靶点。中央 假设是通过2型cysLTR失调的cysLT产生和信号传导 （CysLT 2 R）在引发2型炎症和启动MC中起近端作用。 呼吸道粘膜通过产生IL-33和激活肺 内皮细胞这项提案的结果将对AERD产生直接影响， 其中cysLT是过量产生的，并且通过1型拮抗作用不能完全治疗， cysLTR、CysLT1R。 在目的1中，我们将研究CysLT 2 R在驱动IL-33产生和2型IL-33表达中的作用。 使用新的无效菌株在两种小鼠模型中的肺部炎症。我们将验证 在cysLT驱动的2型炎症中对IL-33的需求，并鉴定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Cysltr 2flox/flox菌株， 鉴定调节这些模型的相关CysLT 2 R表达细胞。在目标3中，我们将使用 来自鼠AERD模型的分选的肺MC的转录谱分析，以检查 CysLT 2 R/IL-33信号传导对激发肺MC产生类花生酸的影响， 病理生物学功能我们将使用在几种条件下定义的鼠MC转录组 为了了解直接从鼻息肉组织中分选的MCs的转录谱， 成人AERD和对照组（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).