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.

项目摘要 T2R苦味受体是最初在舌头上鉴定出的G蛋白偶联受体（GPCR）， 但在其他组织中也起着不同的作用。 “舌外”（舌外）味觉受体可能是 在气道感染，哮喘，前劳动甚至癌症中重要。在气道上皮细胞中，T2RS 4，14， 16和38个识别的企业敏感分子由革兰氏阴性细菌产生，包括 机会性病原体假单胞菌。这些受体的刺激改变了 抗菌分子，包括一氧化氮和防御素。一些多态性降低了功能 T2R的患者增加了患者对上呼吸道感染和慢性鼻炎的敏感性。靶向T2R 由于治疗需要更好地阐明其表达和下游影响。这必须在 原代人类细胞，如小鼠T2R的数量不同，激动剂激活它们。 我们确定了原代人单核细胞和巨噬细胞中的几个T2R受体。巨噬细胞 是吞噬和杀死细菌的重要先天免疫细胞，并分泌促或抗 炎性细胞因子可修饰免疫反应。这些受体的刺激激活钙释放 急性（5-15分钟内）的一氧化氮产生增强了细菌的吞噬作用。初步数据 表明T2RS可能是刺激先天免疫反应以杀死细菌而无需使用的靶标 常规的抗生素，减少了抗生素耐药性的压力。这可能在上层特别有用 呼吸系统疾病。急性和慢性鼻塞炎占所有抗生素处方的20％ 我们，因此是抗生素耐药微生物的重要驱动力。慢性鼻孔炎和 相关的鼻息还会引起许多患者的疾病，缺血和/或厌食症。 我们将进一步定义增加巨噬细胞吞噬作用并测试其如何的信号通路 影响细菌杀死（AIM 1）。因为细菌（例如铜绿假单胞菌）可以侵入细胞并以细胞内生命 病原体逃避了进一步的免疫检测，我们将测试T2RS如何影响巨噬细胞杀菌活性。 这项工作还将是最早使用活细胞成像和荧光生物传感器研究巨噬细胞的工作之一。 GPCR信号实时。接下来（AIM 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（AIM 3）我们将比较表型，T2R表达和T2R功能 来自发炎的慢性鼻腔炎的巨噬细胞鼻息肉，幼稚的血清衍生巨噬细胞来自 同一患者。我们将检查巨噬细胞是否因T2R多态性而改变了息肉或 苦味感知。我们还检查鼻息息巨噬细胞数量是否会影响慢性 鼻孔炎疾病表型。