Anatomical Bases of Human Olfactory Dysfunction

描述（申请人提供）：嗅觉疾病的诊断和治疗的改进首先需要对嗅觉周围的解剖状态进行更全面的分析（鼻中隔和紧邻筛板下方的鼻侧壁上的1cm 2区域）相对于嗅觉上皮（OE）的保存，第二，确定运行经验自身重建的固有能力是否保持完整的一种手段。我们提出了三个具体的目标，旨在解决我们的缺点，外周嗅觉功能障碍的病理基础。目的1：我们观察到一个比预期更好的保存OE在嗅觉区的尸检标本，但不成熟的神经元异常突出。我们将测试的假设，未成熟的神经元占主导地位的地区是断开的OB使用免疫组织化学分析和DiI/DiO追踪轴突。目标二：嗅区OE和RE的混合以及静止期、间歇期OE的发生表明嗅觉干细胞和祖细胞的功能障碍有助于外周嗅觉病理。我们的干细胞和祖细胞在小鼠和大鼠OE的分子概况的研究的指导下，我们将确定用于识别神经干细胞和祖细胞在人类OE的标记。我们将筛选和分析的人类同源基因的表达，是在小鼠使用RT-PCR，免疫组化和原位杂交的人类嗅觉组织活检收集的相关。我们还计划进行开放式基因分析研究。所得到的组可用于测定人嗅觉组织中的干细胞和祖细胞。目的3：检测干/祖细胞标记物和轴突轨迹分析对嗅觉障碍患者的诊断价值。在目标1中嗅觉区研究的指导下，我们将从因衰老、慢性鼻窦炎或先前病毒性URI导致嗅觉障碍的患者中采集嗅觉区活检组织，并使用我们的小组分析干细胞和祖细胞的状态以及组织内轴突生长的模式。公共卫生相关性：在补助金结束时，我们将对尸检时OE的分布和连接性进行深入分析，作为年龄和神经系统疾病的函数，并建立一个诊断小组，可以更清楚地阐明嗅觉功能障碍的病理生理机制。结合起来，所获得的知识可以指导治疗这些疾病的疗法的最终开发。

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. PUBLIC HEALTH RELEVANCE: By the end of the grant we will have an in-depth analysis of the distribution and connectivity of the OE at autopsy as a function of age and neurological disease and established a diagnostic panel that can elucidate more clearly the pathophysiological mechanisms underlying olfactory dysfunction. In combination, the knowledge gained can guide the eventual development of therapies for treatment of these disorders.