Project 1: Mitotic Gene Bookmarking as an Epigenetic Mechanism to Maintain the Mammary Epithelial Phenotype

项目 1：有丝分裂基因书签作为维持乳腺上皮表型的表观遗传机制

基本信息

批准号：
10608053
负责人：
Gary S. Stein
金额：
$ 40.49万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10608053
关键词：
Biological Breast Cancer Cell Breast Cancer Patient Breast Cancer cell line Breast Epithelial Cells CRISPR screen Cause of Death Cell Line Cell Proliferation Cell division Cells ChIP-seq Chromatin Chromosomes Clinical Collaborations DNA Binding Data Development Developmental Process Dimensions Epigenetic Process Epithelial Cell Proliferation Epithelial Cells Epithelium Equilibrium Estrogen Receptors Estrogen receptor positive Estrogens Event Gene Expression Genes Genetic Transcription Genome Genomic Segment Genomic approach Genomics Growth Hi-C Histones Knowledge Link Maintenance Malignant Neoplasms Mammary Neoplasms Maps Mediating Mitosis Mitotic Mitotic Chromosome Modeling Molecular Mutation Neighborhoods Outcome Phenotype Physiological Play Property RUNX1 gene Receptor Signaling Regulation Research Personnel Resolution Role Signal Transduction Specimen Structure Supporting Cell Testing Transcriptional Regulation United States Validation Woman acute myeloid leukemia 1 protein breast cancer progression cancer cell cancer initiation clinically relevant epigenome epithelial to mesenchymal transition histone modification in vivo innovation live cell microscopy loss of function malignant breast neoplasm mammary mammary epithelium novel novel strategies nuclease pharmacologic preservation prevent programs therapeutic target transcription factor transcriptome tumor tumor progression

项目摘要

PROJECT 1 | SUMMARY Mechanisms underlying the epithelial to mesenchymal transition (EMT)—a normal developmental process and an early event at the onset of epithelium-derived cancers—are minimally understood. Our findings, corroborated by others in multiple biologically relevant models, have established mitotic gene bookmarking—occupancy of target genes by transcription factors during mitosis—as a novel dimension to epigenetic control of lineage commitment and cell identity. Because EMT results from loss of the epithelial phenotype, Project 1 will address whether disruption of mitotic gene bookmarking constitutes a key mechanistic component of EMT. Recently, we have shown that depletion of the RUNX1 transcription factor in normal mammary epithelial (NME) cells results in EMT in both in vivo and cell-based models, indicating that RUNX1 stabilizes the normal ME phenotype. Our preliminary data show that in ME cells undergoing mitosis, RUNX1 bookmarks genes involved in cell proliferation, growth and maintenance of the epithelial phenotype. These findings provide the rationale for our hypothesis that mitotic bookmarking of target genes by RUNX1 is a key epigenetic mechanism for maintenance of the normal mammary epithelial phenotype, and perturbation of this mechanism leads to EMT. Innovative molecular and cellular experimental strategies will be combined with state-of-the-art genomics approaches to investigate how the RUNX1 mitotic epigenome contributes to initiation of EMT, an early event that leads to development of breast cancer. Aim 1 will use candidate and unbiased CRISPR screens to determine how RUNX1 bookmarked genes are physiologically linked to the mammary epithelial phenotype and whether estrogen receptor signaling—a key determinant of NME phenotype—plays a role in regulation of RUNX1- bookmarked genes. Aim 2 will employ Hi-ChIP and HIPMap to establish whether RUNX1 bookmarked genes reside in same genomic neighborhoods for coordinate control. We will assess whether disruption of these genomic regions confers EMT. Aim 3 will take advantage of degron-based RUNX1 depletion to investigate the regulatory impact of RUNX1 mitotic bookmarking on target gene expression and maintenance of the NME phenotype. Project 1 studies will address significant gaps in current knowledge of the epigenetic mechanisms that regulate EMT. Our findings will establish RUNX1 mitotic bookmarking as a unique dimension to epigenetic control of the normal mammary epithelial phenotype, which when disrupted, results in EMT, a key early event in the development and progression of breast cancer.

项目1 |概括上皮到间充质转变（EMT）的机制 - 正常的发育过程和在上皮癌开始时的早期事件是最少理解的。我们的发现得到了证实在多个生物学相关模型中，其他人已经建立了有丝分裂基因书签有丝分裂过程中的转录因子靶基因 - 作为谱系表观遗传控制的新维度承诺和细胞身份。由于EMT是由于上皮表型的丢失而导致的，因此项目1将解决有丝分裂基因书签的破坏是否构成EMT的关键机械组成部分。最近，我们已经表明，正常乳腺上皮（NME）中Runx1转录因子的部署细胞在体内和基于细胞的模型中均导致EMT，表明RUNX1稳定了正常的ME 表型。我们的初步数据表明，在我经历有丝分裂的我的细胞中，runx1书签基因涉及在细胞增殖中，上皮表型的生长和维持。这些发现为我们假设Runx1对靶基因的有丝分裂书签是一种关键的表观遗传机制维持正常的乳腺上皮表型和该机制的扰动导致EMT。创新的分子和细胞实验策略将与最先进的基因组相结合研究Runx1有丝分裂遗传组如何促进EMT倡议的方法，EMT的早期事件是导致乳腺癌的发展。 AIM 1将使用候选人和公正的CRISPR屏幕来确定 RUNX1如何将基因与乳腺上皮表型物理联系在一起以及是否存在雌激素受体信号传导（NME表型的关键决定仪）在调节Runx1-的调节中起作用书签基因。 AIM 2将使用Hi-Chip和Hipmap来确定Runx1是否为基因的基因驻留在相同的基因组社区以进行协调控制。我们将评估是否破坏这些基因组区域赋予EMT。 AIM 3将利用基于DEGRON的Runx1部署来调查 RUNX1有丝分裂书签对目标基因表达和NME维持的调节影响表型。项目1研究将解决当前对调节表观遗传机制的知识的差距 EMT。我们的发现将建立Runx1有丝分裂书签，作为对表观遗传控制的独特维度正常的乳腺上皮表型（当被破坏时）导致EMT，这是一个关键的早期事件乳腺癌的发展和进展。