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: 10301509
• 负责人: Jun Nagai
• 金额: $ 25.36万
• 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/10301509
• 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; Cellular Structures; 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; Role; Severities; Severity of illness; Signal Transduction; Signaling Molecule; Source; Specificity; Stimulus; Stress; Stromal Cells; Structure; Structure of mucous membrane of nose; Surface; System; Testing; Therapeutic; Time; Variant; alveolar epithelium; 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.

项目概要 除了作为核酸亚基外，细胞外核苷酸（ATP、ADP、UTP、UDP） 通过激活位于靶细胞质膜上的受体作为生物活性介质。 然而，由于缺乏对它们在许多生物过程和疾病状态中的作用的了解甚少。 我们最近开发了一种全新的高灵敏度生物测定法来检测它们。 我们展示了用于检测生物液体中细胞外核苷酸的系统。 肺部释放的 UDP 促进了针对实验性哮喘的保护性先天免疫反应 小鼠通过肺泡巨噬细胞/IL-12/NK细胞/IFN-γ轴。 长期目标是验证每个核苷酸谱作为有用的诊断或预测 为了实现这一目标，我们将证明该方法的有效性。 对从患有严重哮喘的人类受试者中获得的生物体液中的核苷酸进行定量分析 阿司匹林急性呼吸道疾病（AERD）和相应的动物模型，并确定 我们还寻求扩大我们的生物测定规模，以提供广泛适用的细胞类型。 其他研究人员可以通过该平台检测和监测生物液体中的细胞外核苷酸 在高吞吐量下。 中心假设将通过两个具体目标进行检验：1）确定 患有过敏性炎症性肺部疾病的人类和动物生物体液中的核苷酸谱 疾病的临床特征和严重程度；2) 确定细胞固有的核苷酸释放谱在目标 1 中， 我们寻求每种细胞外核苷酸都具有致病相关性并且可以成为有用的生物标志物 我们将对患有过敏性/炎症性肺部疾病的人类的细胞外核苷酸进行定量和分析。 严重哮喘患者的支气管肺泡灌洗液 (BAL) 分泌物和鼻表面液体 来自 AERD 患者，并确定每个核苷酸与疾病严重程度的相关性，以实现我们的目标 2。 知识，没有细胞类型或刺激特异性特征的全面表征 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）对病理相关刺激做出反应。 这些预期结果和新方法预计将验证每个核苷酸 水平（和核苷酸谱）作为有用的诊断或预测过敏/炎症状态生物标志物。 还有望加深对核苷酸释放机制和哮喘患者的理解 变体，并解决开发基于细胞外核苷酸的治疗策略的限制。