Immune function of bitter taste receptors in human macrophages

人巨噬细胞苦味受体的免疫功能

• 批准号: 10418464
• 负责人: Nithin Adappa
• 金额: $ 51.19万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-25 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10418464
• 关键词: Acute; Affect; Agonist; Airway Disease; Anosmia; Anti-Bacterial Agents; Anti-Inflammatory Agents; Antibiotic Resistance; Antibiotics; Asthma; Bacteria; Biochemical; Biochemistry; Biological Assay; Biosensor; Blood Component Removal; Calcium; Cells; Clinical; Clinical Data; Data; Defensins; Detection; Disease; Disease Progression; Disease model; Dysosmia; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Family; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Genetic Polymorphism; Goals; Gram-Negative Bacteria; Human; Immune; Immune response; Immunologic Receptors; Immunologics; In Vitro; Infection; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Innate Immune Response; Interleukin-4; Invaded; Ions; Link; Malignant Neoplasms; Measurement; Microbiology; Molecular; Mus; Nasal Polyps; Natural Immunity; Neurons; Neutrophil Infiltration; Nitric Oxide; Nitric Oxide Synthase; Nose; Operative Surgical Procedures; Pathway interactions; Patient-Focused Outcomes; Patients; Phagocytosis; Phenotype; Physiology; Polyps; Population; Predisposition; Premature Labor; Production; Pseudomonas aeruginosa; Receptor Signaling; Residual state; Respiratory Disease; Respiratory Tract Infections; Role; Sentinel; Serum; Signal Pathway; Signal Transduction; Sinus; T2R taste receptors; Taste Buds; Taste Perception; Testing; Time; Tissues; Toll-Like Receptor Pathway; Toll-like receptors; Tongue; Upper Respiratory Infections; Work; acute rhinosinusitis; airway epithelium; airway inflammation; antimicrobial peptide; bactericide; base; cGMP production; cell type; chronic rhinosinusitis; combat; cytokine; disease phenotype; eosinophil; experimental study; genotyped patients; hyposmia; immune function; live cell imaging; macrophage; man; microorganism; monocyte; neutrophil; novel; opportunistic pathogen; pathogen; pressure; quorum sensing; receptor; response; therapeutic target; uptake

PROJECT SUMMARY T2R bitter taste receptors are G-protein coupled receptors (GPCRs) originally identified on the tongue, but which also serve diverse roles in other tissues. “Extraoral” (outside the tongue) taste receptors are likely important in airway infection, asthma, pre-term labor, and even cancer. In airway epithelial cells, T2Rs 4, 14, 16, and 38 recognize quorum-sensing molecules produced by gram-negative bacteria, including the opportunistic pathogen Pseudomonas aeruginosa. Stimulation of these receptors modifies production of antibacterial molecules, including nitric oxide and defensins. Some polymorphisms reducing the functionality of T2Rs increase patient susceptibility to upper respiratory infection and chronic rhinosinusitis. Targeting T2Rs as therapies requires better elucidation of their expression and downstream effects. This must be studied in primary human cells, as mouse T2Rs differ in number and agonists that activate them. We identified several T2R receptors in primary human monocytes and macrophages. Macrophages are important innate immune cells that phagocytose and kill bacteria as well as secrete pro- or anti- inflammatory cytokines to modify immune responses. Stimulation of these receptors activates calcium release and nitric oxide production that acutely (within 5-15 min) enhances phagocytosis of bacteria. Preliminary data suggest that T2Rs may be targets for stimulation of innate immune responses to kill bacteria without the use of conventional antibiotics, reducing pressures for antibiotic resistance. This may be especially useful in upper respiratory diseases. Acute and chronic rhinosinusitis account for >20% of all antibiotic prescriptions in the US, and thus are important drives of antibiotic resistant micro-organisms. Chronic rhinosinusitis and associated nasal polyps also cause dysosmia, hyposmia, and/or anosmia in many patients. We will further define the signaling pathway that increases macrophage phagocytosis and test how it affects bacterial killing (Aim 1). As bacteria such as P. aeruginosa can invade cells and live as intracellular pathogens to evade further immune detection, we will test how T2Rs affect macrophage bactericidal activity. This work will also be among the first to use live cell imaging and fluorescent biosensors to study macrophage GPCR signaling in real time. Next (Aim 2), we will examine how T2R activation reduces inflammatory responses to toll-like receptors (TLRs). Our preliminary data suggest novel pathways of T2R-TLR crosstalk possibly involving Akt. Finally (Aim 3) we will compare phenotype, T2R expression, and T2R function in macrophages from inflamed chronic rhinosinusitis nasal polyps with naïve serum-derived macrophages from the same patients. We will examine if macrophage infiltration of polyps is altered with T2R polymorphisms or bitter taste perception. We also examine if increased macrophage numbers in nasal polyps affects chronic rhinosinusitis disease phenotypes.

项目概要 T2R 苦味受体是最初在舌头上发现的 G 蛋白偶联受体 (GPCR)， 但它们也在其他组织中发挥不同的作用。 “口外”（舌头之外）味觉感受器很可能 对于气道感染、哮喘、早产甚至癌症都很重要。在气道上皮细胞中，T2Rs 4, 14, 16和38识别革兰氏阴性细菌产生的群体感应分子，包括 机会致病菌铜绿假单胞菌。刺激这些受体会改变 抗菌分子，包括一氧化氮和防御素。一些减少功能的多态性 T2Rs 增加患者对上呼吸道感染和慢性鼻窦炎的易感性。瞄准 T2R 因为治疗需要更好地阐明它们的表达和下游效应。这个必须要研究一下 原代人类细胞，因为小鼠 T2R 的数量和激活它们的激动剂不同。 我们在原代人单核细胞和巨噬细胞中鉴定了几种 T2R 受体。巨噬细胞 是重要的先天免疫细胞，可以吞噬和杀死细菌以及分泌亲或抗细菌 炎症细胞因子改变免疫反应。刺激这些受体会激活钙的释放 一氧化氮的产生可迅速（5-15 分钟内）增强细菌的吞噬作用。初步数据 表明 T2R 可能是刺激先天免疫反应的目标，可以在不使用 传统抗生素，减少抗生素耐药性的压力。这在上层可能特别有用 呼吸道疾病。急性和慢性鼻窦炎占所有抗生素处方的 20% 以上 美国，因此是抗生素耐药微生物的重要驱动力。慢性鼻窦炎和 相关的鼻息肉还会导致许多患者出现嗅觉障碍、嗅觉减退和/或嗅觉缺失。 我们将进一步定义增加巨噬细胞吞噬作用的信号通路并测试它如何 影响细菌杀灭（目标 1）。由于铜绿假单胞菌等细菌可以侵入细胞并以细胞内的形式生存 为了逃避进一步的免疫检测，我们将测试T2Rs如何影响巨噬细胞的杀菌活性。 这项工作也将是第一个使用活细胞成像和荧光生物传感器来研究巨噬细胞的工作 GPCR 实时信号传导。接下来（目标 2），我们将研究 T2R 激活如何减少炎症 对 Toll 样受体 (TLR) 的反应。我们的初步数据表明 T2R-TLR 串扰的新途径 可能涉及 Akt。最后（目标 3）我们将比较表型、T2R 表达和 T2R 功能 来自发炎的慢性鼻窦炎鼻息肉的巨噬细胞与来自幼稚血清的巨噬细胞 相同的患者。我们将检查息肉的巨噬细胞浸润是否因 T2R 多态性或 苦味感知。我们还检查鼻息肉中巨噬细胞数量的增加是否会影响慢性鼻息肉 鼻窦炎疾病表型。