Anatomical Bases of Human Olfactory Dysfunction

描述(申请人提供)：嗅觉疾病诊断和治疗的改进首先需要更全面地分析嗅觉外周(鼻中隔1平方厘米区域和紧邻筛板下方的鼻侧壁)在保存嗅觉上皮(OE)方面的解剖状况，其次需要确定OE固有的自我重建能力是否保持完好。我们提出了三个具体的目标，旨在解决我们在外周嗅觉功能障碍的病理基础方面的不足。目的1：我们在尸检标本的嗅区观察到比预期更好的OE保存，但未成熟神经元异常突出。我们将通过免疫组织化学分析和DII/DIO轴突追踪来检验这一假设，即未成熟神经元占优势的区域与OB脱节。目的：嗅区OE和RE的混合以及静止性、动脉性OE的发生提示嗅干细胞和前体细胞的功能障碍参与了周围嗅觉的病理过程。在我们对小鼠和大鼠OE中干细胞和祖细胞的分子图谱研究的指导下，我们将寻找识别人类OE中具有神经功能的干细胞和祖细胞的标记。我们将使用RT-PCR、IHC和原位杂交在通过活检收集的人类嗅觉组织上筛选和分析与小鼠相关的基因的人类同源基因的表达。我们还计划进行开放式基因图谱研究。得到的小组可以用来分析人类嗅觉组织中的茎和祖细胞。目的：探讨干/祖细胞标记板和轴突轨迹分析对嗅觉障碍患者的诊断价值。在我们对目标1中嗅觉区域研究的指导下，我们将从因衰老、慢性鼻窦炎或先前的病毒性URI而导致嗅觉障碍的患者身上收集嗅觉区域的活检组织，并使用我们的小组来分析干细胞和祖细胞的状态以及组织内轴突的生长模式。

Description (provided by applicant): Improvements in diagnosis and treatment of olfactory disease will require first, a more comprehensive analysis of the anatomical status of the olfactory periphery (the 1 cm2 areas on the nasal septum and the lateral nasal wall immediately inferior to the cribriform plate) with respect to the preservation of the olfactory epithelium (OE) and, second, a means of determining whether the OE's inherent capacity to reconstitute itself remains intact. We propose three specific aims designed to address our shortcomings with respect to the pathological bases of peripheral olfactory dysfunction. Aim 1: We observe a better than expected preservation of OE in the olfactory area of autopsy specimens, but immature neurons are abnormally prominent. We will test the hypothesis that areas in which immature neurons predominate are disconnected from the OB by using immunohistochemical analysis and DiI/DiO tracing of axons. Aim 2: The intermingling of OE and RE in the olfactory area and the occurrence of quiescent, aneuronal OE indicate that dysfunction of olfactory stem cells and progenitor cells contributes to peripheral olfactory pathology. Guided by our studies of the molecular profile of stem and progenitor cells in mouse and rat OE, we will identify markers for identifying neurocompetent stem and progenitor cells in human OE. We will screen for and analyze the expression of the human homologues of genes that are relevant in mouse using RT-PCR, IHC and in situ hybridization on human olfactory tissue collected by biopsy. We also plan open-ended gene profiling studies. The resulting panel can be used to assay for the stems and progenitors in human olfactory tissue. Aim 3: To test the usefulness of the stem/progenitor marker panel and the analysis of axon trajectory for the diagnosis of dysosmic patients. Guided by our studies of the olfactory area in Aim 1 we will harvest biopsies of the olfactory area from patients rendered dysosmic as a consequence of aging, chronic rhinosinusitis, or prior viral URI, and use our panel to analyze the status of stem and progenitor cells as well as the pattern of axonal growth within the tissue.