Single cell analysis of human olfactory mucosa

嗅觉丧失，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衰退（老年期嗅觉）， 病毒后嗅觉障碍、创伤、炎性损伤或其它特发性病症。目前有 大多数情况下都没有有效的治疗方法。限制战略制定的主要障碍， 预防或治疗厌食症是我们对确切发病机制的有限理解。什么是细胞或 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）来生成定义 细胞群体和状态在成人的嗅觉粘膜。液滴测序技术进展 已经使得这种分析可行，允许从小组织样品中快速分析>5000个细胞。的 鼻内的嗅粘膜是嗅觉的外周器官，容纳了主要的嗅觉感觉器官 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 根据需要我们将把注意力集中在嗅觉粘膜上，因为这是（a）人类中可接近的组织， (b)许多后天性厌食症的可能病理部位。使用scRNA-seq方法，我们将开发 从年轻、中年或高龄的正常成年人的单细胞谱， 了解嗅觉障碍发生的变化所需的基线数据。相比于现有 报告，这是有限的，主要是描述性的，我们提出的实验将提供一个定量图谱， 人类的嗅觉粘膜使用这种方法，我们也将检查嗅觉粘膜的变化 与老视症相关，并检查这是否与涉及神经性 疲惫不堪此外，将制备组织学样本，用于验证以下基因表达模式： 特异性兴趣，使用抗体或RNA原位杂交探针。我们希望R21能够提供基准 数据使未来能够开发针对分子或细胞的诊断和治疗策略 人类嗅觉缺失条件下嗅觉粘膜的变化。

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.