儿童人类嗅上皮在人类嗅器官模型中的结构与功能与成人相似

文献信息

中文摘要

目的： 本研究利用儿童和成人标本以及人类嗅器官，探讨人类嗅上皮（OE）在结构、基因表达和功能方面的发育差异。
背景： 嗅觉功能障碍（OD）以嗅觉丧失或改变为特征，其患病率随年龄增长而增加。小鼠模型已证明嗅上皮中嗅觉受体的表达随时间保持稳定，提示嗅上皮细胞密度可能是年龄相关OD的潜在驱动因素。人类嗅上皮的发育变化仍鲜有表征。在此，我们评估人类嗅上皮在整个生命周期中的结构和功能，以更好地理解正常成熟和年龄相关退化。
方法： 获取儿童和成人受试者的上鼻甲活检组织，用于三维嗅器官培养。进行免疫荧光（IF）和逆转录定量聚合酶链反应（RT-qPCR），以确认预期嗅上皮细胞类型的结构组成和基因表达。进行活细胞钙成像，以评估器官对气味剂的反应，作为功能的替代指标。
结果： IF证实儿童和成人器官包含主要的嗅上皮细胞群（例如，水平基底细胞和球状基底细胞、支持细胞以及未成熟和成熟嗅觉感觉神经元）。儿童和成人组织之间嗅上皮细胞基因表达无统计学差异。钙成像显示气味剂诱发的反应（峰值振幅、曲线下面积）无显著差异。
结论： 在该器官模型所评估的条件下，人类嗅上皮在整个生命周期中似乎在结构、转录和功能上保持稳定。需要进一步研究以更好地理解年龄相关OD，其可能源于非嗅上皮来源的变化。

英文摘要

OBJECTIVE: This study investigated the developmental differences in structure, gene expression, and function of human olfactory epithelium (OE) using pediatric and adult specimens and human olfactory organoids.
BACKGROUND: Olfactory dysfunction (OD) is characterized by loss or alterations in the sense of smell and has an age-related increase in prevalence. Murine models have demonstrated stable expression of olfactory receptors in the OE over time, pointing toward OE cell density as a potential driver of age-related OD. Developmental changes in human OE remain poorly characterized. Herein we assess the structure and function of human OE across the lifespan to better understand normal maturation and age-related degeneration.
METHODS: Superior turbinate biopsies from pediatric and adult subjects were obtained for the culture of three-dimensional olfactory organoids. Immunofluorescence (IF) and reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) were performed to confirm the structural make-up and gene expression of expected OE cell types. Live-cell calcium imaging was performed to assess organoid response to odorants, a surrogate of function.
RESULTS: IF confirmed that pediatric and adult organoids contain major OE cell populations (e.g., horizontal and globose basal cells, sustentacular cells, and immature and mature olfactory sensory neurons). There was no statistical difference in OE cell gene expression between pediatric and adult tissue. Calcium imaging demonstrated no significant difference in odorant-evoked responses (peak amplitude, area under the curve).
CONCLUSION: Human OE appears structurally, transcriptionally, and functionally stable across the lifespan under the conditions assessed in this organoid model. Further research is needed to better understand age-related OD, which may stem from non-OE-derived changes.