Anatomical Bases of Human Olfactory Dysfunction

描述（由申请人提供）：嗅觉疾病的诊断和治疗的改进首先需要更全面地分析嗅觉周围（鼻中隔和鼻侧壁紧挨着筛网板下方的1平方厘米区域）的解剖状态，以及嗅觉上皮（OE）的保存，其次，确定OE的固有重建能力是否保持完整。我们提出了三个具体的目标，旨在解决我们的缺点，尊重周围嗅觉功能障碍的病理基础。目的1：我们在尸检标本的嗅觉区观察到比预期更好的OE保存，但未成熟神经元异常突出。我们将通过使用免疫组织化学分析和轴突的DiI/DiO示踪来验证未成熟神经元占主导地位的区域与OB断开的假设。目的2:OE和RE在嗅觉区域的混合以及静止、动脉瘤性OE的发生表明，嗅觉干细胞和祖细胞的功能障碍参与了周围嗅觉病理。在小鼠和大鼠OE干细胞和祖细胞分子图谱研究的指导下，我们将寻找识别人类OE神经活性干细胞和祖细胞的标记物。我们将使用RT-PCR、免疫组化和原位杂交对活检收集的人类嗅觉组织进行筛选和分析与小鼠相关的人类同源基因的表达。我们还计划进行开放式基因分析研究。所得板可用于测定人嗅觉组织的茎和祖细胞。目的3：测试干细胞/祖细胞标记组和轴突轨迹分析对神经障碍患者诊断的有用性。根据我们在Aim 1中对嗅觉区域的研究，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中收集嗅觉区域的活检，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生长的模式。公共卫生相关性：在拨款结束时，我们将深入分析OE在尸检中的分布和连通性，作为年龄和神经系统疾病的功能，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。综合起来，所获得的知识可以指导治疗这些疾病的治疗方法的最终发展。

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.