Nucleotide releasing profiles in allergic inflammatory diseases

过敏性炎症疾病中的核苷酸释放谱

• 批准号: 10408854
• 负责人: Jun Nagai
• 金额: $ 20.88万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10408854
• 关键词: Address; Allergic; Alveolar Macrophages; Animal Model; Animals; Aspirin; Asthma; Biological; Biological Assay; Biological Markers; Biological Process; Bronchoalveolar Lavage Fluid; Cell Communication; Cell membrane; Cells; Characteristics; Clinical; Complex; Diagnostic; Disease; Effectiveness; Epithelial Cells; Exposure to; Extracellular Fluid; G-Protein-Coupled Receptors; Goals; Human; Immune; Immune response; Inflammatory; Inflammatory Response; Inhalation; Innate Immune Response; Interferon Type II; Interferons; Interleukin-12; Ion Channel; Knowledge; Leukotriene Antagonists; Ligands; Liquid substance; Lung; Lung diseases; Measurement; Mediator of activation protein; Molecular; Monitor; Mus; Natural Killer Cells; Nose; Nucleic Acids; Nucleotides; P2X-receptor; Pathogenesis; Pathologic; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Physiological; Public Health; Pulmonary Pathology; Purinergic P2 Receptors; Pyroglyphidae; Research Personnel; Respiratory Disease; Role; Severities; Severity of illness; Signal Transduction; Signaling Molecule; Source; Specificity; Stimulus; Stress; Stromal Cells; Structure of mucous membrane of nose; Surface; System; Testing; Therapeutic; Time; Variant; alveolar epithelium; antagonist; aspirin-exacerbated respiratory disease; asthmatic patient; base; cell type; detection platform; diagnostic biomarker; driving force; extracellular; human disease; human subject; in vivo; insight; macrophage; novel; novel strategies; predictive marker; receptor; response; therapeutic target

Project Summary In addition to their role as subunits of nucleic acids, extracellular nucleotides (ATP, ADP, UTP, UDP) serve as bioactive mediators by activating receptors localized in the plasma membrane of target cells. However, little is understood about their role in many biologic processes and disease states due to a lack of conventional assay to detect them. We recently developed a completely novel high-sensitivity bioassay system for the detection of extracellular nucleotides in biological fluids. Using this system, we demonstrated that UDP release in lungs facilitated a protective innate immune response against experimental asthma in mice through alveolar macrophage/IL-12/NK cell/IFN-γ axis. The long-term goal is to validate each nucleotide profile as a useful diagnostic or predictive allergic/inflammatory state biomarker. In pursuit of this goal, we will demonstrate the effectiveness of this assay to quantify the nucleotides in biological fluids obtained from human subjects with severe asthma and aspirin-exacerbated respiratory disease (AERD) and corresponding animal models and identify the responsive cell-type. We also seek to expand the scale of our bioassay to provide a broadly applicable platform through which other investigators can detect and monitor extracellular nucleotides in biological fluids at a high throughput. The central hypothesis will be tested by way of two specific aims: 1) To determine the relevance of the nucleotide profile in biological fluids from humans and animals with allergic inflammatory pulmonary diseases to clinical characteristics and severity; 2) To identify cell-intrinsic nucleotide release profiles. In Aim 1, we hypothesize that each extracellular nucleotide is pathogenetically relevant and can be a useful biomarker in humans with allergic/inflammatory pulmonary diseases. We will quantify and profile extracellular nucleotide secretion of bronchoalveolar lavage (BAL) fluids from subjects with severe asthma and nasal surface fluids from patients with AERD, and determine the relevance of each nucleotide to disease severity. In Aim 2, to our knowledge, there is no comprehensive characterization of the cell type- or stimulus-specific profile of nucleotide release. We will characterize nucleotides released by immune (Macrophage) and structural airway cells (Epithelial cells) in response to pathogenetically relevant stimuli. Together, these anticipated results and new approaches are expected to validate each nucleotide level (and nucleotide profile) as a useful diagnostic or predictive allergic/inflammatory state biomarker. Results also promise to deepen the understanding of mechanisms of nucleotides release and asthmatic patient variants, and address the limitation on developing extracellular nucleotide-based therapeutic strategies.

项目摘要 除了它们作为核酸亚基的作用外，细胞外核苷酸（ATP，ADP，UTP，UDP） 通过激活位于靶细胞质膜中的接收器来充当生物活性介质。 但是，由于缺乏 常规测定法以检测它们。我们最近开发了一种完全新颖的高敏性生物测定 使用此系统，我们证明了 在肺中释放的UDP释放制备了针对实验性哮喘的受保护的先天免疫反应 通过肺泡巨噬细胞/IL-12/NK细胞/IFN-γ轴小鼠。 长期目标是验证每个核苷酸概况作为有用的诊断或预测性 过敏/炎症状态生物标志物。为了实现这一目标，我们将证明这一点的有效性 分析以从患有严重哮喘和严重哮喘和 阿司匹林诊断呼吸道疾病（AERD）和相应的动物模型，并确定 响应式细胞类型。我们还寻求扩大生物测定法的规模，以提供广泛的适用 其他研究人员可以通过该平台检测和监测生物流体中的细胞外核苷酸 在高吞吐量。 中心假设将通过两个具体目的进行检验：1）确定相关性 来自人类和动物具有过敏性炎性肺的生物流体的核苷酸谱 临床特征和严重程度的疾病； 2）鉴定细胞内核苷酸释放曲线。在AIM 1中， 我们假设每个细胞外核丁胺在病原上都是相关的，并且可以是有用的生物标志物 在患有过敏/炎症性肺部疾病的人类中。我们将量化和剖面细胞外核苷酸 来自严重哮喘和鼻表面液的受试者的支气管肺泡灌洗（BAL）流体的分泌 来自AERD患者，并确定每种核苷酸与疾病严重程度的相关性。在AIM 2中，我们 知识，没有对细胞类型或刺激特异性的全面表征 核苷酸释放。我们将表征免疫（巨噬细胞）和结构气道释放的核苷酸 细胞（上皮细胞）响应病原体相关的刺激。 总之，这些预期的结果和新方法有望验证每个核苷酸 水平（和核苷酸谱）作为有用的诊断或预测性过敏/炎症状态生物标志物。结果 还有望加深对核苷酸机制释放和哮喘患者的理解 变体，并解决开发细胞外核治疗策略的局限性。