Stromal Epithelial Interactions in Breast Cancer

描述：(申请人提供)建议进行一系列实验，以 测试和验证小鼠乳腺成纤维细胞(Mmgf)将提供 为人类的正常生长和分化创造一个等效的微环境 乳房上皮(HuBrE)。这一观点的确凿证据提出了一种可能性 利用突变小鼠乳腺间质研究旁分泌通路的研究 影响HuBre。此外，我们还将评估 癌相关乳腺成纤维细胞(CAF)对生长、分化、分化的影响 和huBrE的导管形态发生。预计这项研究计划 将导致在细胞和分子水平上的机械论理解 间质-上皮相互作用在生长、分化和导管中的作用 HuBrE的形态发生和基质的变化如何导致初始阶段 易患乳腺癌。这样的研究可能会导致发现新的 乳腺癌患者分层的标志物和可能的新的 治疗。为实现这些目标，提出了5个具体目标。特定的 目的#1分析正常人类乳房的分化和生长 体内生长的上皮细胞与正常来源的成纤维细胞的结合 小鼠和人的乳腺。特定目标2使用组织重组体 突变的小鼠乳腺成纤维细胞与人乳腺上皮细胞构建 评估旁分泌通路在调节生长和分化中的作用 人类乳房上皮的。在这一具体目标中，我们将评估 基质雌激素受体与生长、分化和生长中的“EGF信号” 人乳腺上皮导管的形态发生。具体目标#3是 人乳房分化和生长反应的特征 与癌相关的乳腺成纤维细胞相关的上皮生长 (CAF)小鼠和人类起源，特别关注CAF对(A)的影响 上皮分化和(B)生长反应。具体目标#4侧重于 钙调素对人E-钙粘附素和整合素表达的影响 体内和体外乳腺上皮细胞。我们将具体确定是否 CAF对上皮细胞分化的干扰是由于对 黏附系统，如E-和P-钙粘素、整合素或基底 薄膜。具体目标#5涉及识别独特的分子标记 CAF与上皮基因变化的时程研究 CaF诱导的靶上皮细胞表达。为了实现这些目标，我们 将利用组织重组技术，细胞和器官培养， 免疫细胞化学和各种分子方法学。

DESCRIPTION: (provided by applicant) A series of experiments are proposed to test and validate the idea, that mouse mammary fibroblasts (mMgF) will provide an equivalent microenvironment for proper growth and differentiation of human breast epithelium (huBrE). Definitive proof of this idea raises the possibility of using mutant mouse mammary gland stroma to investigate paracrine pathways affecting huBrE. In addition, we will assess the effects of carcinoma-associated mammary fibroblasts (CAF) on the growth, differentiation, and ductal morphogenesis of huBrE. It is anticipated that this research program will lead to a mechanistic understanding at the cellular and molecular levels of stromal-epithelial interactions in growth, differentiation, and ductal morphogenesis of huBrE and how changes in the stroma lead to the initial stages predisposing to breast cancer. Such research may lead to the discovery of new markers for stratification of breast cancer patients and possibly new therapies. To accomplish these goals, 5 specific aims are proposed. Specific Aim #1 is an analysis of differentiation and growth of normal human breast epithelium grown in vivo in combination with fibroblasts derived from normal mouse and human mammary gland. Specific Aim #2 uses tissue recombinants composed of mutant mouse mammary fibroblasts plus human breast epithelial cells to assess the role of paracrine pathways regulating growth and differentiation of human breast epithelium. In this specific aim we will assess the role of stromal estrogen receptors and "EGF-signaling" in growth, differentiation, and ductal morphogenesis of human breast epithelium. Specific Aim #3 is a characterization of differentiation and growth response of human breast epithelium grown in association with carcinoma-associated mammary fibroblasts (CAF) of mouse and human origin focusing specifically on CAF effects on (a) epithelial differentiation and (b) growth response. Specific Aim #4 focuses on the effects of CAF on E-cadherin (E-cad) and integrin expression in human breast epithelium in vivo and vitro. We will specifically determine whether perturbation of epithelial differentiation by CAF is due to perturbation of adhesion systems such as E- and P-cadherin, intergrins or the basement membrane. Specific Aim #5 deals with identification of molecular markers unique to CAF and an investigation of the time course of change in epithelial gene expression elicited by CAF in target epithelium. To accomplish these goals we will use tissue recombinant technology, cell and organ culture, immunocytochemistry, and various molecular methodologies.