Single cell analysis of human olfactory mucosa

人类嗅粘膜的单细胞分析

基本信息

批准号：
10226794
负责人：
Bradley J Goldstein
金额：
$ 20.1万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10226794
关键词：
Address Adult Affect Age Aging Alzheimer&apos s Disease Anosmia Antibodies Atlases Attention Basal Cell Basic Science Biological Assay Biological Markers Biopsy Biopsy Specimen Cells Computing Methodologies Data Data Set Development Diagnostic Dimensions Disease Elderly Etiology Exhibits Failure Foundations Future Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Diseases Goals Graph Housing Human Impairment In Situ Hybridization Incidence Inflammatory Laboratories Longevity Maintenance Molecular Molecular Target Mucous Membrane National Health and Nutrition Examination Survey Neurodegenerative Disorders Neuroepithelial Cells Neuroglia Neurons Nose Olfactory Epithelium Olfactory Mucosa Olfactory Pathways Olfactory dysfunction Organ Parkinson Disease Pathogenesis Pathology Patients Peripheral Phenotype Population Positioning Attribute Process RNA Reporting Research Scientist Sensory Site Smell Perception Statistical Data Interpretation Supporting Cell Surgeon Techniques Testing Therapeutic Tissue Sample Tissues Transcript Translating Trauma Validation age related base blind cell type clinical practice design droplet sequencing effective therapy exhaustion experimental study genome-wide histological specimens hyposmia immunohistochemical markers interest middle age novel novel therapeutics older patient olfactory disorder olfactory neurogenesis olfactory receptor olfactory sensory neurons potential biomarker prevent progenitor programs receptor expression single cell analysis single-cell RNA sequencing stem stem cells targeted treatment therapy development treatment strategy

项目摘要

PROJECT SUMMARY: Olfactory sensory losses, termed hyposmia or anosmia, are estimated to affect >12% of the US population. Common causes of acquired anosmias include aging-related declines (presbyosmia), post-viral olfactory disorder, trauma, inflammatory damage, or other idiopathic conditions. At present, there are no effective treatments for most of these conditions. A major barrier limiting the development of strategies to prevent or treat anosmias is our limited understanding of the precise pathogenesis. What are the cellular or molecular changes in the olfactory system associated with sensory loss? To address this issue, we propose experiments to apply the single cell RNA sequencing approach (scRNA-seq) to generate datasets defining the cell populations and states within adult human olfactory mucosa. Advances in droplet sequencing techniques have made such assays feasible, permitting the rapid analysis of >5000 cells from a small tissue sample. The olfactory mucosa in the nose is the peripheral organ for olfaction, housing the primary olfactory sensory neurons, related supporting populations, and basal stem cells supporting replacement of neuroepithelial cells as needed. We will focus attention to the olfactory mucosa, since this is (a) an accessible tissue in humans and (b) a likely site of pathology in many acquired anosmias. Using the scRNA-seq approach, we will develop single cell profiles from normosmic adults at young, middle or advanced age to develop comprehensive baseline data necessary for understanding the changes that occur in olfactory disorders. In contrast to existing reports, which are limited and largely descriptive, our proposed experiments will provide a quantitative atlas of the human olfactory mucosa. Using this approach, we will also examine the changes in the olfactory mucosa associated with presbyosmia, and examine whether this is consistent with a process involving neurogenic exhaustion. In addition, histologic samples will be prepared for the validation of gene expression patterns of specific interest, using antibodies or RNA in situ hybridization probes. We expect this R21 to provide baseline data enabling the future development of diagnostic and therapeutic strategies targeting molecular or cellular changes in the olfactory mucosa of human anosmic conditions.

项目摘要：嗅觉感觉丧失（称为嗅觉减退或嗅觉丧失）估计影响 >12% 的美国人口。获得性嗅觉丧失的常见原因包括与衰老相关的嗅觉衰退（老花症）、病毒后嗅觉障碍、创伤、炎症损伤或其他特发性疾病。目前，有大多数这些病症都没有有效的治疗方法。限制制定战略的主要障碍预防或治疗嗅觉丧失是我们对确切发病机制的有限了解。什么是蜂窝或嗅觉系统的分子变化与感觉丧失有关？为了解决这个问题，我们建议实验应用单细胞 RNA 测序方法 (scRNA-seq) 生成定义成人嗅粘膜内的细胞群和状态。液滴测序技术的进展使此类测定变得可行，可以快速分析小组织样本中超过 5000 个细胞。这鼻子中的嗅粘膜是嗅觉的外周器官，具有初级嗅觉感觉神经元、相关支持群体和支持神经上皮细胞替代的基底干细胞根据需要。我们将重点关注嗅觉粘膜，因为这是（a）人类可接近的组织，并且 (b) 许多获得性嗅觉丧失症的可能病理部位。使用 scRNA-seq 方法，我们将开发来自年轻、中年或老年正常成年人的单细胞图谱，以开发全面的了解嗅觉障碍发生的变化所需的基线数据。与现有的相比报告有限且主要是描述性的，我们提出的实验将提供一个定量图谱人类的嗅粘膜。使用这种方法，我们还将检查嗅粘膜的变化与老花症相关，并检查这是否与涉及神经源性的过程一致精疲力尽。此外，将制备组织学样本以验证基因表达模式特定兴趣，使用抗体或RNA原位杂交探针。我们预计 R21 能够提供基准数据有助于未来开发针对分子或细胞的诊断和治疗策略人类嗅觉丧失状况下嗅粘膜的变化。