Role of Chromatin in Defining Breast Epithelial Cell Phenotypes

染色质在定义乳腺上皮细胞表型中的作用

• 批准号: 8050120
• 负责人: KORNELIA POLYAK
• 金额: $ 22.9万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8050120
• 关键词: Biological Assay; Breast; Breast Cancer Cell; Breast Carcinoma; CD44 gene; Cell physiology; Cells; Chromatin; Clinical; DNA; DNA Methylation; Diagnostic Neoplasm Staging; Disease; Distant; ERBB2 gene; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Epigenetic Process; Epithelial; Epithelial Cells; Gene Expression; Genes; Goals; Heterogeneity; Histones; Human; In Vitro; Malignant Epithelial Cell; Malignant Neoplasms; Mammary Gland Parenchyma; Mammary Neoplasms; Mammary gland; Methylation; Molecular Profiling; Mutation; Normal Cell; Outcome; Pathogenesis; Pathologic; Pattern; Phenotype; Play; Population; Principal Investigator; Property; Relapse; Risk; Role; Stem cells; Testing; Tumor Subtype; Tumor stage; base; cell type; chromatin immunoprecipitation; disorder subtype; genome wide association study; in vivo; malignant breast neoplasm; neoplastic; neoplastic cell; progenitor; programs; small hairpin RNA; stem; therapy resistant; transcription factor; tumor; tumor initiation; tumor progression

Distinct normal cells-of-origin may underlie the phenotypic heterogeneity of human breast carcinomas. Perturbations of the normal epigenetic programs of various cells-of-origin may play roles in tumor initiation, metastatic progression, and resistance to therapies. To begin investigating these issues, we characterized. the comprehensive DMA methylation and gene expression profiles of differentiated CD24+ luminal epithelial and CD44+ stem cell-like cells from normal and neoplastic human breast tissue. We identified cell-typespecific DMA methylation patterns that correlated strongly with the histopathological subtypes and clinical outcome of human breast carcinomas. The relatedness of normal and neoplastic DNA methylation patterns was dependent on breast tumor subtypes. Based on these observations, we hypothesize that genes associated with distinct chromatin patterns in mammary epithelial stem and differentiated cells play key roles in determining stem-cell function and lineage-specific differentiation, and that perturbation of these patterns alters cell fate. Furthermore, we hypothesize that normal cell type-specific chromatin patterns are perturbed in breast tumors and that they vary depending on breast tumor types. We propose three specific aims to test these hypotheses. Aim 1: To characterize the histone methylation profiles of human mammary epithelial stem and differentiated cells isolated from normal human breast tissue. We will use ChIP (chromatin immunoprecipitation) and quantitative PCR followed by ChlPSeq genome-wide studies. Aim 2: To characterize the histone methylation profiles of stem cell-like and more differentiated breast cancer cells isolated from different breast tumor subtypes. We will analyze the histone methylation of stem cell-like and more differentiated cells-from different stage tumors of several subtypes (e.g., luminal, HER2+, and basallike tumors) as described in Aim 1. Aim 3. To determine the functional relevance of the cell type-specific epigenetic patterns. To determine the functional relevance of cell type-specific histone methylation patterns, we will analyze the consequences of perturbing these patterns on cell fate and differentiation both in normal and neoplastic human breast tissue.

不同的正常细胞来源可能是人类乳腺癌表型异质性的基础。 各种起源细胞的正常表观遗传程序的扰动可能在肿瘤的发生中发挥作用， 转移进展和对治疗的耐药性。为了开始调查这些问题，我们进行了表征。 分化的 CD24+ 管腔上皮的综合 DMA 甲基化和基因表达谱 以及来自正常和肿瘤性人类乳腺组织的 CD44+ 干细胞样细胞。我们确定了细胞类型特异性 DMA 甲基化模式与组织病理学亚型和临床密切相关 人类乳腺癌的结果。正常和肿瘤 DNA 甲基化模式的相关性 取决于乳腺肿瘤亚型。基于这些观察，我们假设基因 与乳腺上皮干和分化细胞中不同的染色质模式相关，发挥关键作用 确定干细胞功能和谱系特异性分化，以及这些模式的扰动 改变细胞命运。此外，我们假设正常细胞类型特异性染色质模式受到干扰 在乳腺肿瘤中，它们根据乳腺肿瘤类型而变化。我们提出了三个具体的测试目标 这些假设。目标 1：表征人乳腺上皮细胞的组蛋白甲基化谱 从正常人乳腺组织中分离出的干细胞和分化细胞。我们将使用 ChIP（染色质 免疫沉淀）和定量 PCR，然后进行 ChlPSeq 全基因组研究。目标 2： 表征干细胞样和分化程度更高的乳腺癌细胞的组蛋白甲基化谱 从不同的乳腺肿瘤亚型中分离出来。我们将分析干细胞样和干细胞的组蛋白甲基化 更多分化的细胞 - 来自几种亚型的不同阶段肿瘤（例如，luminal、HER2+ 和 basallike） 目标 1. 目标 3. 确定细胞类型特异性的功能相关性 表观遗传模式。为了确定细胞类型特异性组蛋白甲基化模式的功能相关性， 我们将分析扰乱这些模式对正常细胞命运和分化的影响 和肿瘤性人类乳腺组织。