Mechanisms and Receptors Responsible for Leukotriene E4 (LTE4)-dependant Pathobio

负责白三烯 E4 (LTE4) 依赖性 Pathobio 的机制和受体

• 批准号: 8502616
• 负责人: Yoshihide Kanaoka
• 金额: $ 60.58万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8502616
• 关键词: Agonist; Allergens; Aspirin; Asthma; Basophils; Binding; Blood Platelets; Blood Vessels; Breathing; Bronchoconstriction; Cell physiology; Cells; Collaborations; Cutaneous; Disease; Ear; Edema; Extravasation; Family; G-Protein-Coupled Receptors; Generations; Histamine; House Dust Mite Allergens; Human; IgE; Immune; Immune response; In Vitro; Individual; Indomethacin; Laboratories; Lead; Leukotriene C4; Leukotriene D4; Leukotriene E4; Ligands; Lung; Lung diseases; Mediating; Mediator of activation protein; Modeling; Mucous Membrane; Mus; Ovalbumin; Passive Cutaneous Anaphylaxis; Pathology; Pathway interactions; Peroxidases; Pertussis Toxin; Pneumonia; Process; Property; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Pulmonary Eosinophilia; Pulmonary Pathology; Pyroglyphidae; Relative (related person); Resistance; Role; Signal Transduction; Skin; Swelling; Testing; Th2 Cells; Therapeutic; Therapeutic Effect; Tissues; Vascular Permeabilities; Wild Type Mouse; airway hyperresponsiveness; base; clopidogrel; cysteinyl-leukotriene; cytokine; desensitization; eosinophil; eosinophilic inflammation; in vivo; insight; mast cell; member; novel; novel therapeutics; pyroglyphid; receptor; respiratory; response

Individuals with aspirin exacerbated respiratory disease (AERD) demonstrate selective airway hyperresponsiveness to leukotriene (LT)E4 by unclear mechanisms. This Project tests the role of a novel LTE4 receptor, GPR99, in mediating the potent effects of LTE4 on cutaneous and pulmonary vascular leak in mice, and in mediating Th2-dominated pulmonary inflammaton in a model of dust mite-induced pulmonary pathology by a pathway involving platelets, P2Y12 receptors, and prostanoids. We found that mice lacking both CysLT1R and CysLT2R (Cysltr1/Cysltr2-/- mice) show markedly increased and prolonged vascular permeability responses to intradermally administered LTE4 relative to wild-type (WT) controls, as well as an enhanced pulmonary vascular leak in response to LTE4. Cysltr1/Cysltr2-/- mice also show cutaneous and pulmonary vascular leak to LTC4 and LTD4 that are equivalent to those in WT mice. Moreover, Cysltr1/Cysltr2-/- mice show a markedly enhanced and prolonged vascular permeability response in IgE-dependent PCA, indicating that CysLTER mediates ear swelling occurring in response to the cysteinyl leukotrienes (cys-LTs) released by activated mast cells. Finally, intranasal sensitization and challenge with house dust mite extract elicits substantial cys-LT-dependent pulmonary inflammation in Cysltr1/Cysltr2-/- mice, implying that cys-LT-driven immune cell functions that lead to a Th2 cytokine profile and pulmonary eosinophilia can occur independently of CysLT1R or CysLT2R. We have identified GPR99 as a candidate receptor for LTE4 based on preliminary in vitro studies. Aim 1 is to further characterize GPR99 in vitro and to generate the deficient strains needed to prove that GPR99 is the LTE4-specific receptor that mediates vascular and cellular innate and adaptive immune responses in mice lacking CysLT1R and CysLT2R. Aim 2 is to determine the contribution of GPR99 to the cys-LT-dependent mast cell-mediated vascular leakage in the skin of Cysltr1/Cysltr2-/- mice. Aim 3 is to determine the contribution of GPR99 to the cys-LT-dependent allergen-induced Th2 cell mediated pulmonary eosinophilic inflammation in Cysltr1/Cysltr2-/- mice. We will also determine the expression of GPR99 in cells from individuals with AERD and aspirin-tolerant controls.

阿司匹林急性呼吸道疾病（AERD）患者表现出选择性气道 对白三烯（LT）E4的高反应性，其机制尚不清楚。这个项目测试了一部小说的作用 LTE 4受体GPR 99介导LTE 4对皮肤和肺血管渗漏的有效作用 在小鼠中，以及在介导尘螨诱导的肺炎症模型中， 通过涉及血小板、P2 Y12受体和前列腺素类的途径引起病理学改变。我们发现缺乏 CysLT 1 R和CysLT 2 R（Cysltr 1/Cysltr 2-/-小鼠）均显示出显著增加和延长的血管内皮生长因子（VEGF）表达。 相对于野生型（WT）对照，对皮内施用的LTE 4的渗透性反应，以及 增强的肺血管渗漏响应于LTE 4。Cysltr 1/Cysltr 2-/-小鼠也显示皮肤和 肺血管渗漏至LTC 4和LTD 4，相当于WT小鼠中的那些。此外，委员会认为， Cysltr 1/Cysltr 2-/-小鼠在IgE依赖性血管通透性反应中表现出显著增强和延长。 PCA，表明CysLTER介导响应于半胱氨酰半胱氨酸引起的耳肿胀。 由活化的肥大细胞释放的白三烯（cys-LTs）。最后，鼻内致敏和激发 屋尘螨提取物在Cysltr 1/Cysltr 2-/-中激发大量cys-LT依赖性肺部炎症 小鼠，这意味着cys-LT驱动的免疫细胞功能导致Th 2细胞因子谱和肺 嗜酸性粒细胞增多症可以独立于CysLT 1 R或CysLT 2 R发生。我们已经将GPR 99确定为候选者 基于初步体外研究的LTE 4受体。目的1是进一步在体外表征GPR 99， 产生证明GPR 99是LTE 4特异性受体所需的缺陷菌株， 缺乏CysLT 1 R和CysLT 2 R的小鼠的血管和细胞先天性和适应性免疫应答。目的2 是为了确定GPR 99对Cys-LT依赖性肥大细胞介导的血管渗漏的贡献， Cysltr 1/Cysltr 2-/-小鼠的皮肤。目的3是确定GPR 99对cys-LT依赖性细胞凋亡的贡献。 Cysltr 1/Cysltr 2-/-小鼠中变应原诱导的Th 2细胞介导的肺嗜酸性粒细胞炎症。我们将 还测定了来自AERD个体和阿司匹林耐受对照的细胞中GPR 99的表达。