Molecular Function of Cyclin D1 in Breast Development and Breast Cancer

本项目的目标是确定细胞周期蛋白D1在乳腺发育和发育中的分子功能。 N例乳腺癌。细胞周期蛋白D1是核心细胞周期机制的一个组成部分。记录最好的功能 细胞周期蛋白D1的主要功能是结合并激活细胞周期蛋白依赖的CDK4和CDK6。此外, 细胞周期蛋白D1被认为发挥CDK不依赖的功能，可能是细胞周期不依赖的功能，通过作为一个 ：O-特定转录因子的激活物或抑制物。15%-20%的人扩增出细胞周期蛋白D1基因 Uiman乳腺癌，而Cyclin D1蛋白在大多数人乳腺中过表达 癌症。在过去，我们使用细胞周期蛋白D1基因敲除和敲入小鼠来演示对 细胞周期蛋白D1在正常乳腺发育和乳腺肿瘤中发挥作用。分子的精确函数 细胞周期蛋白D1在这些过程中的作用目前还不清楚。这项提议的目的是阐明 细胞周期蛋白D1‘S分子在正常乳腺发育和乳腺肿瘤发生中的作用致信地址 这一期，我们最近产生了一种新的敲入小鼠品系，表达tandemy标记版本的 细胞周期蛋白D1。这种菌株允许我们使用顺序的免疫亲和纯化，然后是高灵敏度 喷枪质谱仪测序，以确定Cyclin D1的全范围相互作用伙伴 在任何发育阶段和癌症进展的任何阶段。我们已经演示过了 这种普遍的方法是成功的，我们现在将其应用于乳腺和 乳腺癌。我们将把我们的分析与对人类乳腺癌的研究结合起来。合并后的 这些方法的使用将使我们能够确定细胞周期蛋白D1在正常乳腺中的分子功能 这将有助于理解这些功能在乳腺肿瘤中是如何错乱的。这个 目的1：研究细胞周期蛋白D1在乳腺发育过程中的分子功能。目标2： 目的：研究细胞周期蛋白D1在乳腺肿瘤发生中的分子功能。 与公共健康相关的Cyclin D1蛋白在许多人类癌症中过表达，包括 大多数乳腺癌患者。尽管细胞周期蛋白D1的过度表达在卵巢癌发病机制中起关键作用 乳腺癌已经确立，细胞周期蛋白D1在这一过程中的确切分子功能仍然存在 不清楚。本申请中提出的工作将解决这一关键问题。对细胞周期蛋白D1‘S的认识 在肿瘤形成中的作用是设计合理的治疗策略所必需的。

The goal of this project is to define the molecular function of cyclin D1 in mammary gland development and n breast cancer. Cyclin D1 is a component of the core cell cycle machinery. The best-documented function of cyclin D1 is its ability to bind and to activate the cyclin-dependent kinases CDK4 and CDK6. In addition, cyclin D1 was proposed to play CDK-independent, possibly cell cycle-independent functions by acting as a :o-activa1or or -represser of specific transcription factors. The cyclin D1 gene is amplified in 15-20% of uiman mammary carcinomas, whereas cyclin D1 protein is overexpressedin the majority of human breast cancers. In the past, we used cyclin D1 knockout and knock-in mice to demonstrate the requirement for cyclin D1 function in normal breast development and in breast neoplasia. The exact molecular function of cyclin D1 in these processes is currently unclear. The goal of this proposal is to elucidate the full range of cyclin D1's molecular functions in normal breast development, and in mammary tumorigenesis. To address this issue, we recently generated a novel knock-in strain of mice expressing tandemly-tagged version of cyclin D1. This strain allows us to use sequential immunoaffinity purifications followed by high-sensitivity shot-gun mass spec sequencing to define the full range of cyclin D1 interacting partners in essentially any organ, at any point of development, and at any stage of cancer progression. We have already demonstrated that this general approach is successful, and we will now apply it to analyses of mammary glands and mammary carcinomas. We will combine our analyses with studies of human breast cancers. The combined use of these approaches will allow us to determine the molecular functions of cyclin D1 in normal breast development, and will help to understand how these functions are deranged in breast neoplasia. The Specific Aims are: Aim 1: To study the molecular function of cyclin D1 during mammary development. Aim 2: To study the molecular function of cyclin D1 during mammary neoplasia. RELEVANCE TO PUBLIC HEALTH Cyclin D1 protein is overexpressed in many human cancers, including the majority of breast cancers. Although the key role for cyclin D1 overexpression in pathogenesis of mammary carcinomas is firmly established, the exact molecular function of cyclin D1 in this process remains unclear. The work proposed in this application will address this critical issue. Understanding of cyclin D1's function in neoplasia is essential in order to design rational therapeutic strategies.