High-throughput Human Cell-Based Models for Primary Prevention of Breast Cancer

用于乳腺癌一级预防的高通量人体细胞模型

基本信息

批准号：
8513576
负责人：
Brittney-Shea Herbert
金额：
$ 9.1万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8513576
关键词：
Accounting Acinus organ component Alcohols Antineoplastic Agents Apical Architecture Behavior Biological Assay Biopsy Birds Breast Breast Cancer Risk Factor Categories Cell Culture Techniques Cell Cycle Cell Line Cell Nucleus Cell model Cells Characteristics Chemistry Chemopreventive Agent Chromatin Collaborations DNA Methylation Development Diet Disease Engineering Environment Epigenetic Process Epithelial Epithelium Event Extracellular Matrix Food Future Gene Expression Genes Genetic Genetic Risk Genetic Transcription Genomics Goals Heart Heterochromatin Homeostasis Human Incidence Life Light Maintenance Malignant Neoplasms Mammary Gland Parenchyma Mammary Neoplasms Methods Microscopy Modeling Modification Molecular Mutation Nuclear Nutrient Obesity Patients Pilot Projects Prevention Prevention strategy Preventive Preventive screening Primary Prevention Proteins Research Risk Risk Assessment Risk Factors Shapes Signal Transduction Source Stratification Stress System Technology Tight Junctions Tissues Tumor Suppressor Proteins Woman Work base cancer cell cancer initiation cancer risk chromatin remodeling design epigenetic marker high risk high throughput screening histone modification improved innovation interest malignant breast neoplasm meetings mutation carrier neoplastic cell new technology prevent promoter public health relevance repository response restoration screening small molecule success tumor tumor initiation

项目摘要

DESCRIPTION (provided by applicant): Primary prevention of breast cancer requires using models of human non-neoplastic cells to decipher the mechanisms of tumor initiation and to design much needed preventive strategies. The HMT3522 S1 non- neoplastic cells mimic the physiologically relevant breast epithelial development by forming ball-shaped three- dimensional (3D) polarized (basal/apical) glandular units (acini) when cultured in contact with an appropriate extracellular matrix (3D culture). Using this model we have identified the loss of apical polarity as a necessary event for cell cycle entry. This suggests that apical polarity is likely to be a ver early architectural modification reflecting an increased risk for tumor onset. Environmental and genetic/epigenetic risk factors for breast cancer have been seldom studied in relevant cell culture models. An important direction of research is to develop cell lines from women with heightened breast cancer risk to unravel the architectural and epigenetic determinants of breast tumor development. Therefore, two research teams for this project are sharing their complementary expertise in the phenotypical analysis of cells in 3D culture and the development and genomic analysis of lines of non-neoplastic cells from women at low or high breast cancer risk. Our goal is to develop high-throughput human cell-based models for the screening of markers of risk assessment and preventive agents. The innovative idea is to use cell lines that represent different breast cancer risks to identify epigenetic markers for elevated risk by correlating these markers to a weakened acinar architecture (i.e., apical polarity loss). The rationale for focusing on epigenetic markers, principally histone modifications as a proof of principle, is that epigenetics is at the heart of gene expression control that goes awry in cancer. Moreover, epigenetic mechanisms are strongly influenced by breast cancer risk factors. To identify meaningful epigenetic markers of breast cancer risk, in Aim 1 we will assess differences in histone modifications between cell lines from low and high breast cancer risk contexts. These modifications will be investigated at the level of nuclear domains known to participate in replication and differentiation (e.g., telomeric and pericentromeric chromatin), and at genes important for epithelial homeostasis. Markers of interest will be validated on archival breast tissue biopsy sections of women at different breast cancer risk levels. In Aim 2, we will determine the architectural and epigenetic responses of the same cell models to dietary and chemically-based modulators of breast cancer risk in order to establish the functionality of the models. Throughout the project we will use a unique high-throughput (HTP) 3D culture method. Future developments would make use of the markers identified and the HTP 3D culture systems to decipher cancer initiation mechanisms and for large scale screening of compound repositories for potential preventive agents using emerging microscopy-based state-of-the-art technologies.

描述（由申请人提供）：主要预防乳腺癌需要使用人类非塑性细胞模型来破译肿瘤起始的机制并设计急需的预防策略。 HMT3522 S1非肿瘤细胞通过形成与适当的细胞外矩阵（3D培养物）接触的培养时，通过形成圆形的三维（基底/根尖）腺体单位（基底/根尖）腺体单位（基底/根尖）腺体单位（基底/根尖）腺体单元（ACINI）来模仿生理相关的乳房上皮发育。使用此模型，我们确定了顶极性的丧失是细胞周期进入的必要事件。这表明顶极性可能是早期的建筑修饰，反映出肿瘤发作的风险增加。在相关细胞培养模型中很少研究乳腺癌的环境和遗传/表观遗传危险因素。研究的一个重要方向是发展乳腺癌风险升高的女性的细胞系，以揭示乳腺肿瘤发育的建筑和表观遗传决定因素。因此，该项目的两个研究团队在3D培养细胞的表型分析以及对低乳腺癌或高乳腺癌风险的女性的非肿瘤细胞线的发展和基因组分析中共享其互补专业知识。我们的目标是开发高通量的人类细胞模型，以筛选风险评估和预防剂的标记。创新的想法是使用代表不同乳腺癌风险的细胞系来识别表观遗传标记以升高通过将这些标记与弱化的腺泡体系结构相关联（即顶极性丧失）来风险。专注于表观遗传标记的基本原理，主要是组蛋白修饰作为原理的证明，是表观遗传学是癌症中出现问题的基因表达控制的核心。此外，表观遗传机制受到乳腺癌危险因素的强烈影响。为了确定乳腺癌风险的有意义的表观遗传标志物，在AIM 1中，我们将评估来自低乳腺癌风险和高乳腺癌风险环境中细胞系之间的组蛋白修饰的差异。这些修饰将在已知参与复制和分化的核域的水平上进行研究（例如，端粒和周围玻璃粒染色质），以及对上皮稳态重要的基因。感兴趣的标记将在不同乳腺癌风险水平的妇女的档案乳房组织活检部分上得到验证。在AIM 2中，我们将确定相同细胞模型对乳腺癌风险的基于饮食和化学的调节剂的结构和表观遗传反应，以建立模型的功能。在整个项目中，我们将使用独特的高通量（HTP）3D培养方法。未来的发展将利用所鉴定的标记和HTP 3D培养系统来破译癌症的发起机制，并使用基于新兴显微镜的先进技术来对复合库的大规模筛选，以供潜在的预防剂筛选。