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：
8628810
负责人：
Brittney-Shea Herbert
金额：
$ 7.52万
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 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8628810
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）：乳腺癌的一级预防需要使用人类非肿瘤细胞模型来破译肿瘤发生的机制并设计急需的预防策略。 HMT3522 S1 非肿瘤细胞在与适当的细胞外基质（3D 培养物）接触培养时，通过形成球形三维（3D）极化（基底/顶端）腺单位（腺泡）来模拟生理相关的乳腺上皮发育。使用这个模型，我们已经确定顶端极性的丧失是细胞周期进入的必要事件。这表明顶端极性可能是一种非常早期的结构修饰，反映了肿瘤发病风险的增加。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从乳腺癌风险较高的女性中开发细胞系，以揭示乳腺肿瘤发展的结构和表观遗传决定因素。因此，该项目的两个研究团队正在分享他们在 3D 培养细胞表型分析以及低乳腺癌风险或高乳腺癌风险女性非肿瘤细胞系的开发和基因组分析方面的互补专业知识。我们的目标是开发基于高通量人类细胞的模型，用于筛选风险评估和预防药物的标记物。创新的想法是使用代表不同乳腺癌风险的细胞系来识别升高的表观遗传标记。通过将这些标记与减弱的腺泡结构（即顶端极性丧失）相关联来评估风险。关注表观遗传标记（主要是组蛋白修饰作为原理证明）的基本原理是，表观遗传学是癌症中出错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有意义的表观遗传标记，在目标 1 中，我们将评估低乳腺癌风险环境和高乳腺癌风险环境中细胞系之间组蛋白修饰的差异。这些修饰将在已知参与复制和分化的核结构域（例如端粒和着丝粒周围染色质）水平以及对上皮稳态重要的基因上进行研究。感兴趣的标记物将在不同乳腺癌风险水平的女性的存档乳腺组织活检切片上进行验证。在目标 2 中，我们将确定相同细胞模型对乳腺癌风险饮食和化学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。未来的发展将利用已确定的标记和 HTP 3D 培养系统来破译癌症引发机制，并使用新兴的基于显微镜的最先进技术大规模筛选潜在预防药物的化合物库。