Solitary chemosensory cell development and function

孤立化学感应细胞的发育和功能

• 批准号: 10194193
• 负责人: Vijay Ramakrishnan
• 金额: $ 5.49万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-05 至 2021-10-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10194193
• 关键词: Address; Agonist; Allergic; Area; Behavior; Calcium; Cell Communication; Cell Count; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cell physiology; Cells; Cellular biology; Chemical Structure; Chemicals; Confocal Microscopy; Data; Detection; Development; Disease; Elements; Environment; Epithelial; Epithelial Cells; Family; Gene Transfer; Goals; Health; Heterogeneity; Homeostasis; House Dust Mite Allergens; Human; Immune response; Immunofluorescence Immunologic; Immunology; Incidence; Inflammatory; Interleukin-13; Irritants; Kinetics; Knowledge; Label; Laboratories; Ligands; Lung; Mentors; Methodology; Methods; Mission; Modeling; Molecular; Morphology; Mucosal Immunity; Mus; Neuroendocrine Cell; Population Heterogeneity; Publications; Quality of life; Quantitative Reverse Transcriptase PCR; Reporter; Research; Rodent; Role; Sensory; Signal Transduction; Structure of respiratory epithelium; System; TRPM5 gene; Taste Buds; Taste Perception; Testing; Time; Tissues; Transgenic Mice; Translating; United States National Institutes of Health; Work; airborne allergen; airway epithelium; airway surface liquid; base; burden of illness; cell type; cigarette smoke; cohesion; confocal imaging; cytokine; disability; effective therapy; epithelial repair; human disease; human study; human tissue; in vivo; innovation; insight; interest; microbial; mouse model; novel; novel strategies; pollutant; precursor cell; progenitor; promoter; receptor; respiratory; response; response to injury; single-cell RNA sequencing; taste transduction; tool; transcriptome sequencing

PROJECT SUMMARY Novel approaches to airway epithelial study have broadened our understanding of its diverse cellular makeup, developmental regulators, response to injury, and cell interactions involved in local tissue function. Recent work by our group and others has demonstrated a role for airway solitary chemosensory cells (SCCs) in detection of irritants within the airway surface liquid, signal transduction through canonical bitter taste signaling effectors, and initiation of a multifaceted immune response. Interestingly, lineage implications from RNAseq based approached in mouse and human lung studies suggest a possible relationship between SCCs and other rare airway cell types including the ionocyte and the neuroendocrine cell. Our central hypothesis is that human SCCs encompass a heterogeneous population that is modulated in the allergic inflammatory setting, and uniquely express cell-specific chemosensory receptors to detect human disease-associated compounds. This hypothesis has been formulated on the basis of publications and preliminary data produced in the applicant's laboratory during the K23 mentored research period, and Co-I Vladar's long-standing interest in airway epithelial repair. To test this—and generate tools for further mechanistic study—we will utilize a platform of retroviral gene transfer to genetically modify SCCs in human cell culture. We will pursue two aims: (1) characterization of SCC differentiation using the hTRPM5p-GFP lentiviral reporter in normal and allergic human primary sinonasal epithelial cell culture; and (2) test human SCC functional responses to disease-associated airborne irritants using a real-time TRPM5-mCherry/GCaMP6 Ca2+ reporter in primary HSNEC culture. Our long-term goal is to determine the functional role(s) of SCCs and chemosensory cell types in airway epithelial homeostasis, and to translate these findings into a cohesive understanding of airway mucosal immunity in human health and disease. Notably, limitations of available methods for labeling and isolating this rare cell type from human tissues have resulted in challenges in translating findings from mouse to human. This innovative proposal represents a significant conceptual departure from the status quo, and will utilize novel methodologies to test specific hypotheses and for general discovery in an area where little is known. The proposed research is significant because it is expected to advance understanding of how SCCs function as environmental sensory that regulate epithelial homeostasis. Ultimately, such knowledge has the potential to be developed into effective therapies for inflammatory airway disorders, a pressing need given the significant incidence and burden of these diseases.

项目总结 呼吸道上皮细胞研究的新方法拓宽了我们对其不同细胞的理解 组成、发育调节、对损伤的反应以及与局部组织功能有关的细胞相互作用。 我们小组和其他人最近的工作证明了呼吸道孤立性化学感觉细胞(Scc)的作用。 在检测呼吸道表面液体中的刺激物时，通过典型的苦味进行信号转导 信号效应器，以及启动多方面的免疫反应。有趣的是，来自 基于RNAseq的小鼠和人肺研究提示鳞状细胞之间可能存在关系 以及其他罕见的呼吸道细胞类型，包括离子细胞和神经内分泌细胞。 我们的中心假设是，人类的干细胞包含了一个异质的种群，这种种群受到 过敏性炎症环境，并独特地表达细胞特异性化学感觉受体来检测人类 与疾病相关的化合物。这一假设是在各种出版物的基础上提出的 申请人的实验室在K23指导研究期间产生的初步数据，以及Co-I Vladar对呼吸道上皮修复的长期兴趣。为了测试这一点-并生成工具以进一步 机制研究-我们将利用逆转录病毒基因转移平台对人类的干细胞进行基因改造 细胞培养。我们将追求两个目标：(1)用hTRPM5P-GFP表征鳞状细胞分化 慢病毒在正常人和变态反应性人鼻腔上皮细胞培养中的表达；以及(2)测试人 实时Trpm5-mCherry/GCaMP6对疾病相关气载刺激物的SCC功能反应 原代HSNEC培养中有钙离子报告。 我们的长期目标是确定干细胞在呼吸道中的功能作用(S)和化学感觉细胞类型 上皮动态平衡，并将这些发现转化为对呼吸道粘膜的凝聚力理解 免疫在人类健康和疾病中的作用。值得注意的是，可用于标记和隔离该病毒的方法的局限性 人类组织中罕见的细胞类型导致了将研究结果从小鼠转化为人类的挑战。 这一创新的提议在概念上与现状有很大的不同，并将利用 测试特定假设和在一个鲜为人知的领域中进行普遍发现的新方法。这个 拟议的研究具有重要意义，因为它有望促进对干细胞如何发挥作用的理解 调节上皮细胞动态平衡的环境感官。最终，这样的知识有可能成为 发展成为治疗炎症性呼吸道疾病的有效疗法，这是一项紧迫的需求，因为 这些疾病的发病率和负担。