Isolation and Characterization of ER+ Breast Cancer Cells with High Bone Metastat

具有高骨转移的 ER 乳腺癌细胞的分离和表征

• 批准号: 8883438
• 负责人: JANET L FUNK
• 金额: $ 7.65万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8883438
• 关键词: Accounting; Address; Automobile Driving; Basal Cell; Biology; Breast Cancer Cell; Breast Cancer Treatment; Breast Cancer cell line; Breast Cancer survivorship; Cause of Death; Cell Line; Cell model; Cells; Clinical; Data Set; Dependency; Development; Diagnosis; Estrogen Antagonists; Estrogen Receptor Status; Estrogen Therapy; Estrogen receptor positive; Estrogens; Female; Funding; Funding Mechanisms; Future; Gene Expression; Genes; Genomic approach; Goals; Health; Human; Laboratories; Lytic; Lytic Lesion; MCF7 cell; Malignant Neoplasms; Metastatic Neoplasm to the Bone; Microarray Analysis; Modeling; Neoplasm Metastasis; Nude Mice; Osteolysis; Osteolytic; Outcome; Parathyroid Hormone Receptor; Pathogenesis; Phenotype; Population Heterogeneity; Pre-Clinical Model; Prevention approach; Primary Neoplasm; Recurrence; Relapse; Reporting; Research; Role; Seminal; Signal Transduction; Site; Specificity; Staging; T47D; Testing; Therapeutic; Tropism; Tumor Cell Biology; Woman; Xenograft Model; anticancer research; base; bone; cell behavior; cell type; clinically relevant; experience; functional genomics; improved; in vivo; innovation; interest; malignant breast neoplasm; model development; neoplastic cell; novel; outcome forecast; parathyroid hormone-related protein; prevent; research study; success; survivorship; tool; treatment strategy; tumor; tumor progression

DESCRIPTION (provided by applicant): Estrogen receptor-positive (ER+) luminal breast cancers, which account for 75% of all breast cancer cases, are the subtype of breast cancer most likely to metastasize to bone, forming incurable lytic lesions that are, in a majority of case, also ER+. In contradistinction, prevailing pre-clinical models investigating the pathogenesis and treatment of breast cancer bone metastases (B-MET) employ ER- basal subtypes of human breast cancer cells. In the absence of standard models of ER+ luminal B-MET, basic questions regarding the biology of these tumor cells within the bone microenvironment cannot be queried, including a possible role for tumor cell estrogen signaling in promoting metastasis progression within the bone microenvironment. The objective of this proposal is to utilize in vivo selection and commonly studied ER+ luminal human breast cancer cell lines to establish robust pre-clinical models of ER+ lytic B-MET which will then be queried, using functional genomics, to examine the pathogenesis of bone metastasis in this setting, comparing and contrasting these findings with those previously obtained in ER- basal cell models. In Specific Aim 1, distinct subpopulations of ER+ cells with a high metastatic potential for formation of lytic B-MET will be isolated by in vivo selection following intracardiac (IC) inoculation of heterogeneous populations of "low-metastatic" human ER+ luminal breast cancer cells (MCF-7, ZR-75-1, or T47D) into nude mice. These studies will take advantage of, and build upon, unique osteotropic ER+ MCF-7-derived cell lines already isolated by our laboratory using this approach. In Specific Aim 2, functional genomic studies, comparing gene expression in cells specifically selected for their bone tropism during the final stage of metastatic progression (osteotropic cells, as described in aim 1) vs. "low- metastatic" cells from which they are derived, will be undertaken to identify genes whose constitutive or estrogen-stimulated expression is associated with tumor progression and osteolysis in the bone metastatic "niche". In vivo testing to determine estrogen-dependency and site-specificity of enhanced tumor progression for these osteotropic cells will further aid in interpreting the functional significance of genes associated with the osteolytic ER+ luminal B-MET phenotype. Success in achieving these aims is likely given that: 1) we are recapitulating the experimental approach used to determine the pathogenesis of ER- basal cell breast cancer B-MET, and 2 ) we have already isolated osteotropic ER+ MCF-7 cells and identified osteolytic B-MET in nude mice inoculated IC with ER+ T47D cells in preliminary experiments. The significance of these studies lies in their novelty, as ER+ luminal models of lytic B-MET have not been developed for interrogation in functional genomic studies, and their clinical relevance to breast cancer, as these studies address a common clinical situation for which there is no cure. Our ultimate goal is to increase survivorship in women diagnosed with breast cancer by improving strategies to prevent and treat ER+ luminal breast cancer B-MET through the current and future use of the novel tools and information acquired in the R03 studies proposed here.

描述（由申请人提供）：雌激素受体阳性（ER+）腔内乳腺癌占所有乳腺癌病例的75%，是最容易转移到骨的乳腺癌亚型，形成无法治愈的溶性病变，大多数情况下也是ER+。相比之下，研究乳腺癌骨转移（B-MET）发病机制和治疗的主流临床前模型采用了人乳腺癌细胞的ER-基底亚型。在缺乏ER+腔内B-MET标准模型的情况下，无法查询这些肿瘤细胞在骨微环境中的生物学基本问题，包括肿瘤细胞雌激素信号在促进骨微环境中转移进展中的可能作用。本研究的目的是利用体内选择和普遍研究的ER+腔型人乳腺癌细胞系建立强大的ER+溶解B-MET临床前模型，然后使用功能基因组学来研究骨转移的发病机制，并将这些发现与先前在ER-基底细胞模型中获得的结果进行比较和对比。在Specific Aim 1中，将异质“低转移”人ER+腔内乳腺癌细胞（MCF-7、ZR-75-1或T47D）在心内（IC）接种裸鼠后，通过体内选择分离出具有形成溶解性B-MET的高转移潜力的不同ER+细胞亚群。这些研究将利用并建立在我们实验室已经用这种方法分离出的独特的促骨性ER+ mcf -7衍生细胞系的基础上。在特异性目标2中，功能基因组研究比较了在转移进展的最后阶段（目标1中描述的嗜骨性细胞）特异性选择的细胞中的基因表达。它们来源于“低转移性”细胞，将用于鉴定其构成性或雌激素刺激表达与骨转移“生态位”中肿瘤进展和骨溶解相关的基因。体内测试以确定这些促骨细胞的雌激素依赖性和增强肿瘤进展的位点特异性，将进一步有助于解释与溶骨ER+相关基因的功能意义