Gene Expression Profiling of Adult Olfactory Stem and Progenitor Cells

描述（由申请人提供）：嗅觉上皮（OE）在损伤后从解剖学和功能上恢复到接近正常的能力，这种能力贯穿整个生命，在神经系统中是独一无二的。然而，我们对嗅觉系统及其下游祖先的分子表型几乎一无所知，这种无知严重阻碍了我们分离它们、理解它们的调节或以一种可控制的方式使用它们的能力。我们建议进行探索性和发展性研究，以解决我们知识上的巨大差距。我们将分离三种特定类型的球状基底细胞（GBCs），在其功能异质的群体中，发现了一种广泛的多能性，干细胞或干细胞样细胞：多能性GBCs，支撑细胞形成的GBCs和过境扩增的GBCs。不同种类的GBCs将根据甲基溴损伤小鼠OE后它们重新出现的时间和它们在一组基本螺旋-环-螺旋（bHLH）转录因子中的一个的表达来定义，这些转录因子包括Hes1和Mash1。表达bHLH TF的细胞将通过FACS分离，根据其表面标记将其定义为GBCs，并与每个单独的TF平行表达GFP（来自转基因或内源基因位点）。对于每个tf定义的群体，我们将进行AffyMetrix微阵列分析，以描述其基因表达模式。此外，正常OE中有丝分裂静止和BrdU标记保留的GBCs是潜在的干细胞群体。我们也将用流式细胞仪分离它们，并对它们进行基因分析。作为这些研究的结果，我们将对静止和激活的嗅觉干细胞及其下游祖细胞有一个全面的了解，并将准备开始假设驱动的功能调节途径评估。OE的干细胞在成人中很容易获得，并且具有制造神经元的自然倾向。此外，嗅觉基板的胚胎细胞在分子和功能上与成体干细胞相似，可以产生专门的嗅觉神经胶质，这可能促进脊髓损伤后的功能恢复。这里收集的信息可能最终使我们能够使用这些成体干细胞来治疗神经系统损伤和退行性疾病。

DESCRIPTION (provided by applicant): The capacity of the olfactory epithelium (OE) to recover both anatomically and functionally after injury to near-normal, a capacity that extends throughout life, is unique in the nervous system. However, we know next to nothing about the molecular phenotype of the olfactory stems nor their downstream progenitors, an ignorance that severely hampers our ability to isolate them, understand their regulation, or use them in a controllable manner. We are proposing exploratory and developmental studies designed to address that vast gap in our knowledge. We will isolate three specific types of globose basal cells (GBCs), among whose functionally heterogeneous population, a broadly pluripotent, stem or stem-like cell is found: multipotent GBCs, sustentacular cell- forming GBCs, and transit-amplifying GBCs. The different kinds of GBCs will be defined by the timing of their re-emergence after methyl bromide lesion of the mouse OE and by their expression of one among a group of basic Helix-Loop-Helix (bHLH) transcription factors that will include Hes1 and Mash1. The bHLH TF-expressing cells will be isolated by FACS on the basis of markers on their surface that define them as GBCs and the expression of GFP in parallel with each individual TF (from either a transgen or the endogenous gene locus). For each TF-defined population we will carry out AffyMetrix microarray analysis designed to profile its pattern of gene expression. In addition, mitotically quiescent and BrdU label-retaining GBCs in the normal OE are a potential stem cell population. We will isolate them by FACS as well and subject them to gene profiling. As a consequence of these studies, we will have a comprehensive understanding of resting and activated olfactory stem cells and their downstream progenitors and will be poised to begin hypothesis-driven assessments of functional regulatory pathways. The stem cells of the OE are both easily accessible in adult humans and have a natural propensity to make neurons. Furthermore, the embryonic cells of the olfactory placode, which the adult stem cells resemble in both molecular and functional terms, give rise to the specialized glia of the olfactory nerve, which may promote functional recovery after spinal cord injury. The information gathered here may allow us eventually to use these adult stem cells in treating nervous system damage and degenerative disease.