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: 8876541
• 负责人: Yoshihide Kanaoka
• 金额: $ 55.46万
• 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/8876541
• 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; In Vitro; Individual; Indomethacin; Laboratories; Lead; Leukotriene C4; Leukotriene D4; Leukotriene E4; Ligands; Lung; Mediating; Mediator of activation protein; Modeling; Mucous Membrane; Mus; Ovalbumin; Passive Cutaneous Anaphylaxis; Pathology; Pathway interactions; Peroxidases; Pertussis Toxin; Process; Property; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Pulmonary Eosinophilia; Pulmonary Inflammation; Pulmonary Pathology; Pyroglyphidae; Relative (related person); Resistance; Role; Signal Transduction; Skin; Swelling; Testing; Th2 Cells; Therapeutic; Therapeutic Effect; Tissues; Vascular Permeabilities; Wild Type Mouse; adaptive immunity; airway hyperresponsiveness; aspirin-exacerbated respiratory disease; asthmatic; 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.

患有阿司匹林加重呼吸道疾病 (AERD) 的个体表现出选择性气道 对白三烯 (LT)E4 的高反应性机制尚不清楚。这个项目测试了小说的作用 LTE4 受体 GPR99 介导 LTE4 对皮肤和肺血管渗漏的有效作用 在小鼠中，以及在尘螨诱发的肺部模型中介导 Th2 主导的肺部炎症 通过涉及血小板、P2Y12 受体和前列腺素的途径进行病理学研究。我们发现小鼠缺乏 CysLT1R 和 CysLT2R（Cysltr1/Cysltr2-/- 小鼠）均显示血管显着增加和延长 相对于野生型（WT）对照，皮内注射 LTE4 的渗透性反应，以及 LTE4 响应的肺血管渗漏增强。 Cysltr1/Cysltr2-/- 小鼠也表现出皮肤和 LTC4 和 LTD4 的肺血管渗漏与 WT 小鼠中的情况相当。而且， Cysltr1/Cysltr2-/- 小鼠在 IgE 依赖性中表现出显着增强和延长的血管通透性反应 PCA，表明 CysLTER 介导响应半胱氨酰而发生的耳朵肿胀 激活的肥大细胞释放白三烯 (cys-LT)。最后，鼻内致敏和挑战 屋尘螨提取物在 Cysltr1/Cysltr2-/- 中引发大量 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可导致 Th2 细胞因子谱和肺部疾病 嗜酸性粒细胞增多可以独立于 CysLT1R 或 CysLT2R 发生。我们已将 GPR99 确定为候选者 基于初步体外研究的 LTE4 受体。目标 1 是进一步表征 GPR99 的体外特性并 产生证明 GPR99 是介导 LTE4 特异性受体所需的缺陷菌株 缺乏 CysLT1R 和 CysLT2R 的小鼠的血管和细胞先天性和适应性免疫反应。目标2 的目的是确定 GPR99 对 cys-LT 依赖性肥大细胞介导的血管渗漏的贡献 Cysltr1/Cysltr2-/- 小鼠的皮肤。目标 3 是确定 GPR99 对 cys-LT 依赖性的贡献 Cysltr1/Cysltr2-/- 小鼠中过敏原诱导的 Th2 细胞介导的肺部嗜酸性粒细胞炎症。我们将 还测定了 AERD 和阿司匹林耐受对照个体细胞中 GPR99 的表达。