Bitter and sweet taste receptor physiology in airway ciliated cells

气道纤毛细胞中的苦味和甜味受体生理学

• 批准号: 10440041
• 负责人: Robert J. Lee
• 金额: $ 4.69万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10440041
• 关键词: Air; Altered Taste; Antibiotic Resistance; Antibiotics; Bacteria; Biochemical; Biochemistry; Biological Assay; Calcium Signaling; Cells; Chronic; Cilia; Data; Diffuse; Dimerization; Disease; Endothelial Cells; Endothelium; Enzymes; Epithelial Cells; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Genetic Polymorphism; HSP 90 inhibition; Heat-Shock Proteins 90; Human; Immune; Immunity; Immunofluorescence Microscopy; Impairment; Infection; Inhalation; Irritants; Link; Liquid substance; Lung diseases; Mediating; Molecular; Molecular Chaperones; Mucociliary Clearance; Nasal Epithelium; Natural Immunity; Nitric Oxide; Nitric Oxide Synthase; Nose; Operative Surgical Procedures; Organ; Outcome; Paint; Parents; Pathway interactions; Phagocytosis; Pharmacology; Phosphorylation; Phosphotransferases; Physiological Processes; Physiology; Play; Portraits; Predisposition; Prevalence; Production; Protein Isoforms; Proteins; Quinolones; Receptor Activation; Receptor Signaling; Research; Residual state; Respiratory Tract Infections; Role; Signal Pathway; Signal Transduction; Sinus; Site; Superbug; Surface; Taste Bud Cell; Taste Buds; Taste Perception; Testing; Therapeutic; Tissues; Tongue; Upper Respiratory Infections; Virus; airway epithelium; arm; bactericide; base; cell type; chronic rhinosinusitis; cilium motility; enzyme activity; falls; homoserine lactone; inhibitor/antagonist; innate immune pathways; insight; kinase inhibitor; live cell imaging; macrophage; novel; pathogen; protein folding; rat Gnat3 protein; receptor; surgery material; surgery outcome; sweet taste perception

PROJECT SUMMARY T2R bitter taste receptors in airway cell cilia detect bacterial acyl-homoserine lactones and quinolones. Activation of these T2Rs increases nitric oxide (NO) production to increase ciliary beating and kill bacteria. T2Rs thus play an important role in airway innate immunity. Polymorphisms that reduce the function of cilia T2Rs correlate with increased gram negative upper respiratory infections, susceptibility to chronic rhinosinusitis, and worse surgical outcomes after sinus surgery. Targeting T2Rs or enhancing their activity may be a complementary or alternative to antibiotics. Boosting endogenous T2R-mediated immunity in the airway or other organs may be a useful anti-pathogen strategy in the face of the increasing prevalence of antibiotic-resistant “superbugs.” NO can be bactericidal and also can inactivate viruses. T2Rs also activate NO production in other tissues and immune cells like macrophages, where T2R to NO signaling regulates phagocytosis. T2Rs activate the endothelial (e) form of nitric oxide synthase (eNOS) in both primary airway cells and macrophages. This pathway appears to be a hallmark of T2R signaling in many cell types and regulates multiple physiological processes. Understanding how T2Rs signal to eNOS is critical for understanding extraoral taste receptor signaling in many tissues. We hypothesize that T2R to eNOS signaling requires phosphorylation of eNOS by kinases activated downstream of the T2Rs. Kinase pathways activated by T2Rs are very unexplored compared with many other GPCRs. We also hypothesize that this kinase phosphorylation is enhanced by HSP90, an important cellular chaperone protein involved in NOS function as well as some GPCR signaling. In this proposal, we will investigate T2R-mediated phosphorylation of NOS and consequences for NO production in primary nasal epithelial cells isolated from residual surgical material and grown at air-liquid interface (Aim 1). We will use live cell imaging of NO production and biochemistry combined with well-characterized pharmacological inhibitors of kinase pathways known to activate eNOS. We will also use well characterized pharmacological HSP90 inhibitors to characterize if HSP90 regulates T2R or eNOS localization to airway cilia or function (calcium signaling or NO production).

项目概要 气道细胞纤毛中的 T2R 苦味受体检测细菌酰基高丝氨酸内酯 和喹诺酮类药物的激活会增加一氧化氮 (NO) 的产生。 因此，纤毛跳动和杀死细菌在气道先天免疫中发挥着重要作用。 纤毛 T2R 功能降低的多态性与革兰氏阴性菌增加相关 上呼吸道感染、慢性鼻窦炎易感性以及较差的手术结果 鼻窦手术后，靶向 T2R 或增强其活性可能是一种补充或补充。 增强气道或其他部位内源性 T2R 介导的免疫力。 面对日益流行的疾病，器官可能是一种有用的抗病原体策略 NO 可以杀菌，也可以灭活病毒。 T2R 还激活其他组织和免疫细胞（如巨噬细胞）中 NO 的产生， 其中 T2R 至 NO 信号传导调节吞噬作用。 原代气道细胞和巨噬细胞中的一氧化氮合酶（eNOS）。 似乎是许多细胞类型中 T2R 信号传导的标志，并调节多种 了解 T2Rs 如何向 eNOS 发出信号对于理解生理过程至关重要。 我们帮助许多组织中的口外味觉受体信号转导到 eNOS 信号转导。 需要 T2R 下游激活的激酶对 eNOS 进行磷酸化。 与许多其他 GPCR 相比，T2R 激活的途径尚未被探索。 HSP90 增强了该激酶的磷酸化，HSP90 是一种重要的细胞因子 伴侣蛋白参与 NOS 功能以及一些 GPCR 信号传导。 在本提案中，我们将研究 T2R 介导的 NOS 磷酸化和 从手术残留中分离出的原代鼻上皮细胞中 NO 产生的后果 材料并在气液界面生长（目标 1）。我们将使用 NO 产生的活细胞成像。 和生物化学与成熟的激酶药理学抑制剂相结合 我们还将使用已明确药理学的 HSP90 激活途径。 抑制剂来表征 HSP90 是否调节 T2R 或 eNOS 定位至气道纤毛或功能 （钙信号传导或 NO 产生）。