Urologic Epithelial Stem Cells

描述(由申请人提供)： 成人前列腺和膀胱中存在一组具有自我更新能力的上皮祖细胞。在可扩散信号的诱导下，细胞与适当的基质间充质细胞相互作用；前体细胞经历分化，产生功能成熟的细胞。因此，当前列腺诱导基质细胞触发时，来自前列腺或膀胱的祖细胞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识别谱系中的细胞类型。CD分子在器官的组成细胞类型中有不同的表达，因此非常适合于这种分析。我们建议研究前列腺上皮细胞分化的过程，并阐明可能导致细胞异常增殖的遗传障碍，这些疾病具有良性增生和癌症等常见前列腺疾病的特征。我们的目标是：(1)使用CD荧光抗体和激光共聚焦扫描显微镜来显示在不同器官中发现的前体细胞的表型相似，而分化后的细胞在CD表达方面的表型不同，并且可以识别不同的中间细胞类型；(2)使用流式细胞术来表征和分类CD表达的前体细胞(特别是CD49b；CD49f和CD71)，这些前体细胞被认为在这些器官和其他器官中包含相似的比例；(3)通过体外三维细胞培养表明，前列腺形态发生(导管和腺体的形成)可以用祖细胞和基质细胞衍生的因子来模拟，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)，它通过与响应的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)表明功能分化(如腔细胞合成丰富的分泌蛋白前列腺特异性抗原(PSA)所表明的)需要上皮/基质细胞-细胞相互作用；(5)证明CD10和CD13的差异表达在腔上皮细胞分化中起作用，因为肿瘤细胞是腔状的，缺乏CD10和CD13的表达；(6)测定前体细胞的转录组，鉴定前体细胞和分化细胞之间的差异表达基因，筛选由基质细胞表达的诱导前列腺上皮分化的基因；以及(7)建立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.