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: 9521663
• 负责人: Robert J. Lee
• 金额: $ 40.25万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9521663
• 关键词: Acids; Address; Agonist; Air; Airway Disease; Anti-Bacterial Agents; Antibiotics; Apical; Bacteria; Biochemical; Biochemistry; Biological Models; Bronchioles; Calcium; Carrier Proteins; Cell Differentiation process; Cell Line; Cell physiology; Cells; Cellular biology; Cilia; Data; Diffuse; Dimerization; Epithelial; Fluorescence Resonance Energy Transfer; Functional disorder; Funding; G-Protein-Coupled Receptors; GLUT2 gene; Genes; Genetic Polymorphism; Glucose; Glucose Transporter; Goals; Growth; Human; Human Cell Line; Immune; Immune signaling; Immunity; Immunofluorescence Immunologic; Impairment; Individual; Infection; Inhalation; Innate Immune Response; Irritants; Knock-out; Knockout Mice; Knowledge; Light; Liquid substance; Lung; Measurement; Membrane; Metabolism; Modeling; Molecular; Mucociliary Clearance; Mus; Natural Immunity; Nitric Oxide; Nitric Oxide Synthase; Nose; Operative Surgical Procedures; Outcome; Pathway interactions; Patients; Pharmacology; Phosphorylation; Phosphotransferases; Physiology; Play; Predisposition; Production; Protein Isoforms; Proteins; Proteomics; Proto-Oncogene Proteins c-akt; Quinolones; Receptor Signaling; Regulation; Research; Respiratory Tract Infections; Role; SLC2A1 gene; Sensory; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Sinus; Surface; System; T1R receptor; T2R taste receptors; Taste Buds; Taste Perception; Techniques; Testing; Therapeutic; Tissues; Tongue; Translating; Western Blotting; Work; airway epithelium; airway surface liquid; arm; cell type; chronic rhinosinusitis; cilium motility; defense response; glucose transport; glucose uptake; homoserine lactone; inhibitor/antagonist; insight; knock-down; live cell imaging; novel; overexpression; pathogen; receptor; receptor function; response; sweet taste perception; therapeutic target

We recently discovered that human nasal ciliated cells express the T2R bitter taste receptors. When activated by secreted bacterial products, T2Rs in cilia stimulates an innate immune signaling cascade involving calcium-driven nitric oxide production that increases ciliary beating as well as directly kills bacteria. Genetic polymorphisms in the TAS2R38 gene, one of the T2Rs in cilia, may underlie susceptibility to infection in patients with chronic rhinosinusitis. We hypothesize that activation of T2R bitter receptor responses in airway ciliated cells will activate innate immune to help eradicate infections without the use of conventional antibiotics. Understanding how to develop topical therapeutics targeting these pathways requires further knowledge of the identity of other cilia-localized chemosensory receptors in cilia as well as their signaling pathways and downstream effects. There remains a critical need for knowledge of the cell biology and physiology of extraoral taste receptors in general. We hypothesize that sinonasal and bronchial motile cilia express multiple chemosensory receptors, as we have identified multiple T2Rs (4,14,16, and 38) and T1Rs 2 and 3 (components of the sweet taste receptor) localized to sinonasal cilia. Interestingly, T2Rs and T1Rs activate different pathways, in contrast to the tongue where their intracellular signaling is similar. Cilia T2Rs activate Ca2+-dependent NO production, possibly also involving the kinase AKT. Activation of cilia T1R2/3 activates a distinct signaling pathway that regulates airway epithelial glucose transporters. Understanding cilia chemosensation and the unique signaling pathways involved will shed light on how to leverage these receptors for therapeutic benefit as well as elucidate mechanisms of non-canonical taste receptor signaling that may be highly relevant to the tongue and/or the many extraoral tissues where they are expressed. In Aim 1, we will further elucidate the signaling of cilia T2Rs using a combination of live cell imaging, biochemical, and molecular approaches in differentiated primary human cells and cell lines cultured at air-liquid interface. In Aim 2, we will use similar techniques to elucidate the signaling mechanism by which T1R receptors regulate glucose transport in the airway as well as its regulation by T1R receptors in primary human and mouse cells. In Aim 3, we will examine the localization and interactions of T1Rs and T2Rs using a combination of live-cell imaging and biochemistry, as well as identify the cilia chemosensory repertoire by biochemical cilia purification and proteomics. Together, the independent yet inter-related aims will examine important chemosensory functions of airway cilia, revealing new ways to leverage chemosensory receptors as therapeutic targets for airway diseases. Equally importantly, we will reveal new insights into the cell biology of extraoral taste receptor function that will likely translate to T1Rs and T2Rs in other tissues. While much of T2R and T1R cell biology has been inferred from heterologous systems, our unique model system will allow us to study the function and interactions of endogenous T1R and T2R function in differentiated primary human cells.

我们最近发现人鼻纤毛细胞表达 T2R 苦味受体。什么时候 纤毛中的 T2R 被分泌的细菌产物激活，刺激先天免疫信号级联反应，涉及 钙驱动的一氧化氮的产生可增加纤毛的跳动并直接杀死细菌。遗传 TAS2R38 基因（纤毛中的 T2R 之一）的多态性可能是感染易感性的基础 慢性鼻窦炎患者。我们假设气道中 T2R 苦味受体反应的激活 纤毛细胞将激活先天免疫，帮助根除感染，而无需使用传统抗生素。 了解如何开发针对这些途径的局部疗法需要进一步 了解纤毛中其他纤毛定位化学感应受体的身份及其信号传导 途径和下游效应。仍然迫切需要细胞生物学知识和 口外味觉感受器的一般生理学。我们假设鼻窦和支气管运动纤毛 表达多种化学感应受体，因为我们已经鉴定了多种 T2R（4、14、16 和 38）和 T1R 2 3（甜味受体的成分）定位于鼻窦纤毛。有趣的是，T2R 和 T1R 激活不同的途径，与舌头相反，舌头的细胞内信号传导相似。纤毛T2R 激活 Ca2+ 依赖性 NO 产生，可能还涉及激酶 AKT。纤毛 T1R2/3 的激活 激活调节气道上皮葡萄糖转运蛋白的独特信号通路。了解纤毛 化学感觉和所涉及的独特信号通路将揭示如何利用这些受体 以获得治疗益处并阐明可能是非经典味觉受体信号传导的机制 与舌头和/或表达它们的许多口外组织高度相关。 在目标 1 中，我们将结合活细胞成像进一步阐明纤毛 T2R 的信号传导， 气液培养的分化原代人类细胞和细胞系的生化和分子方法 界面。在目标 2 中，我们将使用类似的技术来阐明 T1R 的信号传导机制 受体调节气道中的葡萄糖转运以及原代人类中 T1R 受体的调节 和小鼠细胞。在目标 3 中，我们将使用 活细胞成像和生物化学的结合，以及通过以下方式识别纤毛化学感应库 生化纤毛纯化和蛋白质组学。独立但相互关联的目标将一起审查 气道纤毛的重要化学感应功能，揭示了利用化学感应受体的新方法 气道疾病的治疗目标。同样重要的是，我们将揭示细胞生物学的新见解 口外味觉受体功能可能会转化为其他组织中的 T1R 和 T2R。虽然大部分 T2R T1R 细胞生物学是从异源系统推断出来的，我们独特的模型系统将使我们能够 研究分化的原代人类细胞中内源性 T1R 和 T2R 功能的功能和相互作用。