Normal and Cancer Stem Cells of the Mammary Gland

乳腺的正常干细胞和癌症干细胞

• 批准号: 7895525
• 负责人: Larry Ray Rohrschneider
• 金额: $ 52.97万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-17 至 2011-07-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7895525
• 关键词: A Mouse; Ablation; Address; Adult; Affect; Alveolar; Alveolus; Biological Markers; Cell Separation; Cell model; Cell physiology; Cells; Classification; Collaborations; Data; Development; Ductal; ERBB2 gene; Embryo; Epithelial Cells; Exhibits; Fatty acid glycerol esters; Gene Expression; Gene Expression Profile; Gene Proteins; Generations; Genes; Goals; Green Fluorescent Proteins; Growth and Development function; Human; Human Identifications; Impairment; In Vitro; Investigation; Label; Lobular; Lobule; Malignant Neoplasms; Mammary Neoplasms; Mammary Tumorigenesis; Mammary gland; MicroRNAs; Milk; Molecular; Molecular Profiling; Molecular and Cellular Biology; Mus; Nature; Oncogenes; One-Step dentin bonding system; Population; Pregnancy; Primary Neoplasm; Process; Production; Property; Puberty; Regulation; Research; Research Proposals; Role; Signal Transduction; Specificity; Stem cells; Structure; System; Technology; Testing; Therapeutic Uses; Tissues; Transgenic Mice; Transgenic Model; Translating; Transplantation; Uncertainty; anticancer research; base; cancer stem cell; cancer type; cell growth; cell type; enhanced green fluorescent protein; insight; interest; malignant breast neoplasm; mammary gland development; mouse model; neoplastic cell; notch protein; prospective; public health relevance; research study; self-renewal; stem; stem cell fate; stem cell niche; therapeutic target; tumor

DESCRIPTION (provided by applicant): The major long-term objectives of this research proposal are to characterize the normal mammary tissue stem cells (MaSC) in the mouse, and determine how, and/or whether, MaSC participate in cancer development. These results in the mouse will be compared to human breast tumor types for extrapolating results to the human system. A mouse transgenic model in which the enhanced green fluorescent protein (GFP) is expressed in several stem/progenitor cell populations of the embryo and adult. In the adult mammary tissue, two unique GFP+ stem cell types have been observed during puberty, coincident with mammary ductal outgrowth, and again in early to late pregnancy, prior to the generation of alveolar/lobular structures. The GFP+ cells in puberty correspond to the cap cells in the growing end of the terminal end buds, which form the initial mammary ductal network. In early pregnancy the GFP+ cells appear as alveolar buds, which direct formation of alveoli/lobules necessary for milk production. The prospective labeling with GFP offers a one-step isolation of the GFP+ cell populations. These GFP+ cells are MaSC as demonstrated by transplantation outgrowth, lineage tracing using Cre/LoxP technology, and self- renewal experiments. The research proposed in this application will focus on three specific Aims, which will first, characterize the MaSC niche based on expected niche properties, determine Notch-dependent regulation of MaSC fate, and identify MaSC functions by stem cell ablation experiments. Secondly, the prospective identification of MaSC allows the direct test of the cancer stem cell hypothesis. Experiments in the second Aim will determine whether Wnt1 and ErbB2 (also called neu or HER2) mammary tumors contain GFP+ tumor cells, which contain the primary tumor-initiating activity. In addition, the target cell, within all mammary tissue epithelial cells, will be identified by cell isolation and direct in vitro transformation. Tumor cell readout will follow transplantation into mammary fat pads for outgrowth. The third Aim, will tie together GFP+ murine mammary tumors with gene expression-based human mammary tumor classifications. MicroRNAs in these GFP+ murine mammary tumors will be identified for further analysis of signaling and potential therapeutic use. PUBLIC HEALTH RELEVANCE: The prospective identification and characterization of mammary stem cells has enabled experiments to be performed, which were not previously possible. Therefore, results from this project will generate new insights into mammary stem cells and how they may contribute to normal mammary tissue and to cancer development. Addressing the cancer stem cell hypothesis will clarify present uncertainties in the target cells for which therapies should be directed, while gene expression data will draw parallels between mouse and human mammary cancers. These studies represent a new and promising paradigm for breast cancer research.

描述（由申请人提供）：该研究建议的主要长期目标是表征小鼠中正常的乳腺组织干细胞（MASC），并确定MASC是否参与癌症的发展。将小鼠中的这些结果与人类乳腺肿瘤类型进行比较，以将结果推送到人类系统。小鼠转基因模型，其中增强的绿色荧光蛋白（GFP）在胚胎和成人的几个茎/祖细胞群中表达。在成年乳腺组织中，在青春期期间观察到了两种独特的GFP+干细胞类型，与乳腺导管生长相吻合，在肺泡/小叶结构产生之前，在妊娠早期至后期再次观察到。青春期中的GFP+细胞对应于末端芽生长端的盖细胞，该末端芽形成了初始乳腺导管网络。在怀孕初期，GFP+细胞显示为肺泡芽，直接形成牛奶生产所需的肺泡/小叶。使用GFP的前瞻性标记提供了GFP+细胞群体的一步隔离。这些GFP+细胞是MASC的，如移植生长，使用CRE/LOXP技术的谱系跟踪以及自我更新实验所证明的。本应用程序中提出的研究将集中在三个特定目标上，首先将根据预期的小众特性来表征MASC的特征，确定Notch依赖性MASC命运的调节，并通过干细胞消融实验鉴定MASC功能。其次，MASC的前瞻性鉴定允许直接检验癌症干细胞假设。第二个目标中的实验将确定Wnt1和Erbb2（也称为NEU或HER2）是否含有GFP+肿瘤细胞，其中包含原发性肿瘤炎活性。另外，将通过细胞分离和直接体外转化来鉴定靶细胞，在所有乳腺组织上皮细胞中。肿瘤细胞读数将遵循移植到乳腺脂肪垫中以进行生长。第三个目标将将GFP+鼠乳腺肿瘤与基因表达的人类乳腺肿瘤分类绑在一起。这些GFP+鼠乳腺肿瘤中的microRNA将被确定，以进一步分析信号传导和潜在的治疗用途。公共卫生相关性：乳腺干细胞的前瞻性鉴定和表征已使实验能够进行，这是不可能的。因此，该项目的结果将产生对乳腺干细胞的新见解，以及它们如何促进正常的乳腺组织和癌症的发展。解决癌症干细胞假设的解决将阐明应定向疗法的目标细胞中的当前不确定性，而基因表达数据将在小鼠和人类乳腺癌之间取得平行。这些研究代表了用于乳腺癌研究的新且有希望的范式。