TGFA & TGFB IN PROSTATIC STROMAL/EPITHELIAL INTERACTIONS

转化生长因子A

• 批准号: 2147307
• 负责人: MITCHELL S. STEINER
• 金额: $ 15.44万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1997-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2147307
• 关键词: RNase protection assay; androgens; benign prostate hyperplasia; cell cell interaction; cell cycle; embryogenesis; endoderm; epithelium; gene expression; genetically modified animals; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; laboratory mouse; mesenchyme; mesoderm; messenger RNA; mixed tissue /cell culture; newborn animals; organ culture; protein biosynthesis; reproductive development; seminal vesicles; stromal cells; transforming growth factors

Benign prostatic hyperplasia (BPH) is a clinically significant disease characterized by excessive stromal and epithelial proliferation. This has been proposed to result from reactivation of embryonic mesenchymal- epithelial interactions. androgens are probably not the primary growth regulator because their addition or removal does not uniformly affect adult prostate growth suggesting that other factors are directly responsible for prostate cellular proliferation. Peptide growth factors may be those embryonic inducers responsible for aberrant prostate cellular proliferation leading to BPH. Thus, understanding the normal role of peptide growth factors and androgens in prostate development may provide mechanistic insights concerning the mesenchymal-epithelial interactions that are likely reiterated in BPH. Our working hypothesis is that peptide growth factors are direct mediators of androgen action on mesenchymal-epithelial cellular interactions responsible for prostate development. The adjacent mesoderm-derived seminal vesicle development will be contrasted that of the endoderm-derived prostate. The mouse model will be used to test the hypothesis through these specific aims. Aim 1- To determine the spatial and temporal expressions of TGFalpha and TGFBeta1-TGFBeta3 by mesenchymal and epithelial cells during the critical developmental stages for seminal vesicle and prostate with and without androgens and with transgenic mice that misexpress TGFalpha, TGFBeta, or both TGFalpha and TGFBeta. In addition to morphometric analysis, colocalization of TGFalpha and TGFBeta1-TGFBeta3 protein and mRNA expression of prostate with and without androgens and with transgenic animals will be performed during the three critical developmental stages which are 1) commitment to male structures stages (gestational day 15- 17), 2) ductal elongation stage (gestational day 18-postnatal day 3) and 3) branching morphogenesis androgen-independent (up to postnatal day 12) and androgen-dependent (after postnatal day 14). Aim 2 - To determine if TGFalpha and TGFBeta3 may directly mediate the androgen-dependent development of the prostate and seminal vesicles in the absence of androgens. Two approaches will be used: 1) Separate the prostate into primary pure cultures of mesenchymal and epithelial cells and examine pure cell and mixed cell cultures for elaboration of TGFalpha and TGFBeta with and without androgens, 2) Utilize organ cultures of developing prostate and seminal vesicles to test whether addition of pure exogenous TGFalpha or TGFBeta may directly mimic androgen effects during critical developmental stages. Aim 3 To investigate whether c-myc and n-myc are the molecular mechanisms by which TGFalpha and TGFBeta act on mesenchymal and epithelial cells during development of the prostate and seminal vesicle. This will be approached by conducting in situ hybridization of myc and correlating this with TGFalpha and TGFbeta mRNA and protein expression during the critical stages of development. It is the goal of this proposal to use the mouse model, both wild type and transgenics, to define the fundamental role of androgens and peptide growth factors in the development of the prostate, and thus, to provide important new information about mesenchymal-epithelial interactions that may be the basis for BPH and perhaps, prostatic neoplasia as well.

良性前列腺增生（BPH）是一种临床意义的疾病 以过度的基质和上皮增殖为特征。 这 已经提出是由胚胎间充质的重新激活引起的 上皮相互作用。 雄激素可能不是主要的增长 调节器，因为它们的添加或去除不一致地影响 成人前列腺生长表明其他因素是直接的 负责前列腺细胞增殖。 肽生长因子 可能是负责异常前列腺的胚胎诱导者 细胞增殖导致BPH。 因此，了解正常 肽生长因子和雄激素在前列腺发育中的作用可能 提供有关间质上皮的机械见解 BPH中可能重申的相互作用。 我们的工作假设 是肽生长因子是雄激素作用的直接介体 在负责前列腺的间质上皮细胞相互作用上 发展。 相邻的中胚层的囊泡发育 将与内胚层衍生的前列腺形成对比。 鼠标 模型将通过这些特定目的来检验假设。 目标1-确定tgfalpha的空间和时间表达式 临界期间间充质和上皮细胞的TGFBETA1-TGFBETA3 带有和不带有和没有 雄激素和带有Misexpress tgfalpha，tgfbeta或的转基因小鼠 TGFALPHA和TGFBETA。 除形态分析外， TGFALPHA和TGFBETA1-TGFBETA3蛋白和mRNA的共定位 前列腺的表达和不含雄激素和转基因的表达 将在三个关键发展阶段进行动物 是1）对男性结构阶段的承诺（妊娠天15- 17），2）导管伸长阶段（妊娠第18天第3天）和 3）分支形态发生雄激素独立（直至产后第12天） 和雄激素依赖性（产后第14天之后）。目标2-确定是否 TGFALPHA和TGFBETA3可能直接介导雄激素依赖性 在没有的情况下发展前列腺和精液囊泡 雄激素。 将使用两种方法：1）将前列腺分开 间质和上皮细胞的原代纯培养物，并检查 纯细胞和混合细胞培养物，用于阐述TGFALPHA和TGFBETA 有或没有雄激素，2）利用器官培养 前列腺和精液测试是否添加纯外源性 tgfalpha或tgfbeta可能在关键期间直接模仿雄激素效应 发展阶段。 目标3以调查C-MYC和N-MYC是否为 TGFALPHA和TGFBETA对间充质作用的分子机制 前列腺和精液的发育过程中的上皮细胞 囊泡。 这将通过进行现场杂交来解决 MYC并将其与TGFALPHA和TGFBETA mRNA和蛋白质相关联 在发展的关键阶段的表达。 使用鼠标模型，这是该提议的目的 和转基因，以定义雄激素和肽的基本作用 前列腺发展中的增长因素，从而提供 有关间质上皮相互作用的重要新信息 也许是BPH的基础，也许也是前列腺肿瘤的基础。