Role of taste signaling in airway epithelial function

 描述(申请人提供)：在这个项目中，我们的目标是研究化学感官检测在鼻和鼻窦炎性疾病中的作用。这些疾病--特别是鼻炎和鼻窦炎--是极其普遍和令人不安的疾病。最近，基础科学研究利用特殊细胞中的味觉信号通路建立了啮齿动物呼吸道炎症的机制，这种特殊细胞被称为孤立化学感觉细胞(SCCs)。我们最近在人体组织中发现了这些细胞，我们的长期目标是研究干细胞在人类鼻腔炎症性疾病中的作用。我们的第一个目标是使用从身体和人体组织中获得的免疫染色技术来确定这些细胞在鼻腔中的位置。根据我们的初步实验，我们预计会看到适度的丰度和不均匀的分布。干细胞是人类呼吸道上皮中尚未研究的一组新的细胞，了解这些细胞的分布将有助于将来对它们进行活组织检查。 研究。在第二个目标中，我们将基于解剖位置和疾病状态来检查味觉受体的分布。人类有25种苦味感受器，我们希望看到其中一些更常出现在鼻子前面，另一些会在鼻窦更深的地方发现。同样，有些基因可能在过敏性鼻炎和慢性鼻-鼻窦炎等疾病状态下表达更多。为了验证这一假设，我们将评估这些苦味感受器在呼吸道不同区域的相对表达，并将健康组织与患病患者组织进行比较。我们将测试经典刺激物和病原体与人类患者中识别的受体的对比，以确定这些受体是否能够检测到潜在的刺激物。这些发现将建立化学感官检测与人类鼻炎和鼻窦炎疾病的联系，并为治疗这些高负担的慢性疾病的新疗法打开大门。在第三个也是最终目标中，我们将评估是否需要味觉转导来调节粘膜表面的细菌定植。人与人之间的微生物群差异很大，但影响细菌定植的宿主因素却知之甚少。在啮齿动物和人类身上的先前证据表明，SCC和苦味机制能够检测呼吸道病原体，并启动反应以杀死和清除这些细菌。在这个目标中，我们将使用现代DNA测序技术来研究(1)缺乏使用苦味通路的能力的小鼠和(2)具有遗传苦味受体缺陷(T2R38)的人类的呼吸道表面细菌群落。我们对人类鼻窦的初步研究结果表明，这些通路可能发挥作用；证实这一点将代表着一项真正的创新发现，为化学感觉转导建立了新的相关性。

 DESCRIPTION (provided by applicant): In this project, we aim to study the role of chemosensory detection in inflammatory diseases of the nose and sinuses. These diseases-in particular rhinitis and rhinosinusitis-are extremely prevalent and bothersome disorders. Recently, basic science research has established a mechanism for airway inflammation in rodents using taste signaling pathways in specialized cells known as solitary chemosensory cells (SCCs). We have recently found these cells in human tissues, and our long-term objective is to examine the role of SCCs in human inflammatory diseases of the sinonasal cavity. Our first aim is to determine where these cells are located in the sinonasal cavity, using immunostaining techniques on tissue obtained from cadavers and human subjects. We expect to see a moderate abundance and uneven distribution based on our preliminary experiments. SCCs are a novel population of cells yet to be studied in human respiratory epithelium, and understanding the distribution of these cells will facilitate the ability to biopsy them for future research. In the second aim, we will examine the distribution of taste receptors based on anatomic location and disease state. There are 25 human bitter taste receptors, and we expect to see that some of these are found more often in the front of the nose, and others will be found deeper in the sinuses. Similarly, some may be more expressed in disease states such as allergic rhinitis and chronic rhinosinusitis. To test this hypothesis, we will evaluate the relativ expression of these bitter taste receptors in different areas of the airway, and compare healthy to diseased patient tissues. We will test classic irritants and pathogens against receptors identified in human patients to determine if these receptors are able to detect the potential stimulants. These findings will establish the connection of chemosensory detection to the human diseases of rhinitis and rhinosinusitis, and open the door for novel therapies to treat these highly burdensome chronic diseases. In the third and final aim, we will evaluate if taste transduction is required to regulate the bacterial colonization of the mucosal surface. The microbiome varies markedly between people, yet the host factors that influence bacterial colonization are poorly understood. Prior evidence in rodents and humans has shown that SCCs and bitter taste mechanisms are able to detect respiratory pathogens and initiate a response to kill and clear these bacteria. In this aim, we will use modern DNA sequencing techniques to examine the airway surface bacterial communities from (1) mice that lack the ability to use bitter taste pathways, and (2) humans that have a genetic bitter receptor defect (T2R38). Our preliminary results from human sinuses suggest a role for these pathways; confirmation of this would represent a truly innovative finding that establishes a new relevance for chemosensory transduction.