P190-B RhoGAP regulates the microenvironment in the developing mammary gland

描述(由申请人提供)：我的长期职业目标是获得一个教员职位，并研究调节乳腺(MG)发育的分子途径，最终目标是确定它们在乳腺癌中是如何被破坏的。我的直接目标是利用新的实验方法开发一个独立的研究项目，以提高我的研究技能。虽然这个建议的一些目标是我博士后研究的延伸，但大多数是我可以建立独立研究计划的新的研究路线。罗森博士的实验室和贝勒医学院(BCM)拥有最先进的技术、核心实验室、培训计划，以及与30多名MG生物学家/乳腺癌研究人员高度合作的氛围，以促进我的研究取得成功。因此，我认为这是一个理想的环境，可以在其中追求职业发展奖，这将有助于我过渡到教师职位。 我建立了可诱导的p190-B RhoGAP过表达小鼠，以研究必要的信号通路在MG发展和癌症不同阶段的作用。我证明了p190-B是MG形态发生所必需的，并确定了p190-B在调节控制MG发育的间质-上皮相互作用中的一个新角色。这项建议的目的是研究p190-B调控乳腺上皮细胞(MEC)形态发生的信号相互作用，以及介导MG发生的间质-上皮相互作用。为了在体外模拟MG的形态发生，我将使用成熟的3D培养方法培养从p190-B小鼠分离的原代MEC和基质细胞。这将使我能够剖析复杂的细胞和分子相互作用，p190-B通过这些相互作用调节MG的形态发生，而这是无法在体内完成的。我将在体内检测间质细胞在介导p190-B对MG发育影响中的作用，并将对显微解剖的上皮和间质进行基因芯片分析，以确定p190-B过表达扰乱MG形态发生的基因表达变化。 现在很明显，乳腺肿瘤周围的环境被破坏，这促进了肿瘤的生长和扩散。了解环境如何调节MG的发育是确定其破坏如何促进乳腺癌的关键第一步。P190-B在人类乳腺癌中的作用尚不清楚。由于像p190-B这样对发育至关重要的蛋白质经常在肿瘤发生过程中被破坏，p190-B可能在乳腺癌中发挥重要作用。如果我们要确定p190-B在乳腺癌中的作用，了解p190-B在正常MG发育中的功能是至关重要的。

DESCRIPTION (provided by applicant): My long-term career goal is to obtain a faculty position and to investigate the molecular pathways that regulate mammary gland (MG) development with the ultimate goal of determining how they are disrupted in breast cancer. My immediate goal is to develop an independent research program using new experimental approaches that will enhance my research skills. While a few of the objectives of this proposal are extensions of my postdoctoral studies, the majority are new lines of investigation on which I can build an independent research program. Dr. Rosen's laboratory and Baylor College of Medicine (BCM) have state-of-the-art technology, core laboratories, training programs, and a highly collaborative atmosphere with over 30 MG biologists/breast cancer researchers to facilitate success in my research. Thus, I believe that this is an ideal environment in which to pursue a career development award that will foster my transition to a faculty position. I generated inducible p190-B RhoGAP overexpressing mice to study the role of an essential signaling pathway in distinct stages of MG development and cancer. I showed that p190-B is required for MG morphogenesis and identified a novel role for p190-B in regulating the stromal-epithelial interactions that govern MG development. The aims of this proposal will investigate the signaling interactions by which p190-B regulates mammary epithelial cell (MEC) morphogenesis and the stromal-epithelial interactions that mediate MG development. To model MG morphogenesis in vitro, I will grow primary MECs and stromal cells isolated from the p190-B mice using a well-established 3D culture assay. This will allow me to dissect the complex cellular and molecular interactions by which p190-B regulates MG morphogenesis, which cannot be done in vivo. I will test the role of stromal cells in mediating the effects of p190-B on MG development in vivo, and microarray analysis will be performed on microdissected epithelium and stroma to determine the gene expression changes by which p190-B overexpression disrupts MG morphogenesis. It is now evident that the environment surrounding a breast tumor is disrupted, which facilitates tumor growth and spread. Understanding how the environment regulates MG development is a critical first step in determining how its disruption promotes breast cancer. The role of p190-B in human breast cancer is unknown. Because proteins like p190-B that are essential for development frequently become disrupted during tumorigenesis, p190-B may play an important role in breast cancer. An understanding of p190-B function in normal MG development is crucial if we are to determine how it is involved in breast cancer.