Urologic Epithelial Stem Cells

描述（由申请人提供）： 具有自我更新能力的上皮祖细胞群存在于成人前列腺和膀胱中。由扩散信号和与适当的间质细胞的细胞间相互作用诱导;祖细胞经历分化以产生功能成熟的细胞。因此，来自前列腺或膀胱的祖细胞在被前列腺诱导基质细胞触发时将发育成前列腺结构。对于细胞类型分析，我们将使用簇指定（CD）细胞表面分子的表达来鉴定谱系中的细胞类型。CD分子在器官的组成细胞类型中差异表达，因此非常适合这种分析。我们建议研究前列腺上皮细胞分化的过程，并证明可能的遗传疾病，导致异常细胞增殖的良性增生和癌症的常见前列腺疾病的特点。我们的目标是：（1）使用荧光CD抗体和共聚焦激光扫描显微镜来显示在不同器官中发现的祖细胞在表型上相似，而分化的细胞在表型上就其CD表达而言是不相似的，并且可以鉴定不同的中间细胞类型;（2）使用流式细胞术来通过CD表达表征和分选（特别是，CD 49 b; CD 49 f和CD 71）祖细胞，其被假定在这些和其它器官中包含相似的比例;（3）通过体外三维细胞培养显示，（导管和腺体的形成）可以用祖细胞和基质细胞衍生因子建模以产生分泌腔细胞的分化细胞类型（主要的基质因子之一是肝细胞生长因子（HGF），其通过与反应性上皮细胞上的酪氨酸激酶受体c-met结合来激活形态发生程序）;（4）功能分化（如通过管腔细胞合成丰富的分泌蛋白、前列腺特异性抗原（PSA）所示）需要上皮/基质细胞-细胞相互作用;（5）显示差异表达的CD 10和CD 13外肽酶在管腔细胞分化中起作用，因为管腔样的癌细胞缺乏CD 10和CD 13表达;（6）确定祖细胞的转录组，鉴定祖细胞和分化细胞之间差异表达的基因，筛选诱导前列腺上皮分化的基质细胞表达的基因;和（7）建立所获得的实验结果的公共数据库。

DESCRIPTION (provided by applicant): A population of epithelial progenitor cells with self-renewal capability resides in the adult human prostate and urinary bladder. Induced by diffusible signals, and cell-cell interaction with the proper stromal mesenchyme cells; the progenitor cells undergo differentiation to produce functionally mature cells. Thus, progenitor cells from either the prostate or the bladder will develop into prostatic structures when triggered by prostate-inducing stromal cells. For cell type analysis we will use the expression of cluster designation (CD) cell surface molecules to identify cell types in lineages. CD molecules are differentially expressed among the component cell types of organs, and are therefore ideally suited to such analysis. We propose to study the process of prostatic epithelial cell differentiation and to demonstrate possible genetic disorder that leads to abnormal cell proliferation characteristic of the common prostatic diseases of benign hyperplasia and cancer. Our objectives are: (1) to use fluorescent CD antibodies and confocal laser scanning microscopy to show that the progenitor cells found in diverse organs are phenotypically alike whereas the differentiated cells are phenotypically dissimilar with respect to their CD expression, and that distinct intermediate cell types can be identified; (2) to use flow cytometry to characterize and sort by CD expression (in particular, CD49b; CD49f, and CD71) progenitor cells, which are postulated to comprise a similar proportion in these and other organs; (3) to show by in vitro 3-D cell cultures that prostate morphogenesis (formation of ducts and glands) can be modeled with progenitor cells and stromal cell derived factors to give rise to the differentiated cell type of secretory luminal cells (one of the principal stromal factors is hepatocyte growth factor (HGF), which activates the morphogenetic program through its binding to tyrosine kinase receptor c-met on responsive epithelial cells); (4) to show that functional differentiation (as indicated by the synthesis of the abundant secretory protein, prostate-specific antigen (PSA), by luminal cells) requires epithelial/stromal cell-cell interaction; (5) to show that the differentially expressed CD10 and CD13 ectopeptidases play a role in luminal cell differentiation, since cancer cells, which are luminal-like, lack CD10 and CD13 expression; (6) to determine the transcriptome of progenitor cells, to identify differentially expressed genes between progenitor and differentiated cells, to screen for genes expressed by stromal cells that induce prostate epithelial differentiation; and (7) to build a public database of the experimental results obtained.