Single cell analysis of human olfactory mucosa

人类嗅粘膜的单细胞分析

• 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
• 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' 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

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：嗅觉损失（称为缺血或厌食症）估计会影响> 12％ 美国人口。获得的厌氧症的常见原因包括与衰老相关的下降（Presbymia）， 病毒后嗅觉疾病，创伤，炎症损伤或其他特发性疾病。目前，有 对于大多数情况，没有有效的治疗方法。限制战略发展的主要障碍 预防或治疗厌氧是我们对精确发病机理的有限理解。什么是细胞或 与感觉丧失有关的嗅觉系统的分子变化？为了解决这个问题，我们建议 实验以应用单细胞RNA测序方法（SCRNA-SEQ）生成定义的数据集 成人人类嗅觉粘膜中的细胞种群和状态。液滴测序技术的进步 已经使这种测定可行，从而可以从小组织样品中快速分析> 5000个细胞。这 鼻子中的嗅觉粘膜是用于嗅觉的外围器官 神经元，相关的支持人群和支持替代神经上皮细胞的基础干细胞 根据需要。我们将把注意力集中在嗅觉粘膜上，因为这是（a）人类中可访问的组织 （b）许多获得的厌氧症中可能的病理部位。使用SCRNA-SEQ方法，我们将发展 来自年轻，中年或高龄的正常成年人的单细胞剖面以发展全面 理解嗅觉疾病中发生的变化所需的基线数据。与现有 报告的报告有限且在很大程度上是描述性的，我们提出的实验将提供一个定量地图集 人类嗅觉粘膜。使用这种方法，我们还将检查嗅觉粘膜的变化 与长老会有关，并检查这是否与涉及神经源的过程一致 精疲力尽。另外，将准备组织学样本以验证基因表达模式 使用抗体或RNA原位杂交探针的特定兴趣。我们希望这款R21提供基线 数据实现了针对分子或细胞的诊断和治疗策略的未来发展 人类厌氧条件的嗅觉粘膜的变化。

项目成果

Preparation of Human Olfactory Epithelial Biopsies for Downstream Analysis.

制备用于下游分析的人类嗅觉上皮活检。

• DOI: 10.1007/978-1-0716-3425-7_10
• 发表时间: 2023
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Choi,Rhea;Ko,Tiffany;Finlay,JohnB;Hachem,RalphAbi;Jang,David;Goldstein,BradleyJ
• 通讯作者: Goldstein,BradleyJ