Subnuclear Targeting and Architectural Epigenetics in Cancer Cells

癌细胞的亚核靶向和结构表观遗传学

• 批准号: 8601045
• 负责人: Gary S. Stein
• 金额: $ 25.32万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8601045
• 关键词: Acetylation; Acute Myelocytic Leukemia; Architecture; Biochemical; Biological; Breast Cancer Cell; Cell Fate Control; Cell Nucleus; Cell division; Cells; Chimeric Proteins; Chromatin; Chromatin Structure; Chromosomes; Chromosomes, Human, Pair 1; Collaborations; Commit; DNA Methylation; DNA Polymerase I; DNA Polymerase II; DNA Sequence; Dimensions; Environment; Epigenetic Process; Etiology; Event; Exhibits; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genomics; Growth; Histones; Instruction; Interphase; Link; Malignant Neoplasms; Mediating; Methods; Microscopic; Microscopy; Mitosis; Mitotic; Mitotic Chromosome; Modification; Molecular; Normal Cell; Nuclear; Nuclear Structure; Oncogenic; Phenotype; Post-Translational Protein Processing; Proteins; Proteomics; Regulation; Regulator Genes; Research Personnel; Role; Shapes; Signal Transduction; Somatic Cell; Structure; Variant; cancer cell; cancer stem cell; cell growth; cohort; leukemia; metaplastic cell transformation; novel; programs; promoter; runx proteins; t(8; 21)(q22; q22); trafficking; transcription factor; tumorigenesis

Cancer cells exhibit alterations in parameters of nuclear architecture that control cell fate and compromise control of cell growth. Our Program has established new biological paradigms by showing that gene regulatory factors integrate cell signaling at chromatin microenvironments ('subnuclear foci') and support epigenetic mechanisms through association with mitotic chromosomes. In collaboration with other Program Project investigators. Project 1 will now establish new dimensions in gene regulation by defining perturbations in architecturally linked regulatory mechanisms during interphase and mitosis in AML and breast cancer cells. Our central hypothesis is that (i) subnuclear targeting of transcription factors to gene regulatory foci during interphase and (ii) the association of transcription factors with their target genes in mitotic chromosomes are fundamental to the retention of biological states of normal and cancer cells. Therefore, we will use IF microscopy, biochemical, genomic and proteomic approaches (i) to characterize modifications in architectural epigenetics and molecular pathological consequences of expressing the translocation-related t(8;21) AML-ETO fusion protein (Aim 1), (ii) to analyze genes that are transcriptionally and spatially controlled by Runx2 in chromatin micro-environments ('subnuclear foci') during interphase in breast cancer cells (Aim 2), and (iii) to examine Runx2 mediated architectural epigenetics in breast cancer cells by characterization of Runx2 and cognate gene regulatory factors that associate with mitotic chromosomes (Aim 3). By investigating the functional role of Runx2 in establishing chromatin micro- environments ('subnuclear foci') during interphase and architectural epigenetics in cancer cells during mitosis, we will challenge traditional biochemical views of gene regulation by defining the pathological linkages between modifications in nuclear architecture and gene expression that are fundamental to the molecular etiology of tumorigenesis.

癌细胞表现出核结构参数的变化，这些参数控制着细胞的命运，损害了细胞生长的控制。我们的计划已经建立了新的生物学范式，表明基因调控因子整合了染色质微环境(亚核焦点)的细胞信号，并通过与有丝分裂染色体的关联来支持表观遗传机制。与其他项目项目调查员合作。项目1现在将通过定义急性髓细胞白血病和乳腺癌细胞间期和有丝分裂期间结构上相关的调控机制的扰动，建立基因调控的新维度。我们的中心假设是：(I)转录因子在间期亚核定位于基因调控区；(Ii)转录因子与其靶基因在有丝分裂染色体中的结合是正常细胞和癌细胞保持生物学状态的基础。因此，我们将使用IF显微镜、生化、基因组和蛋白质组学方法(I)表征体系表观遗传学中的修饰和表达易位相关t(8；21)AML-ETO融合蛋白(目标1)的分子病理学后果，(Ii)分析乳腺癌细胞间期在染色质微环境中受Runx2转录和空间调控的基因(‘亚核焦点’)(目标2)，以及(Iii)通过表征Runx2和与有丝分裂染色体相关的同源基因调控因子(目标3)来研究Runx2介导的乳腺癌细胞的体系表观遗传学。通过研究Runx2在建立癌细胞间期染色质微环境(亚核焦点)和有丝分裂期间构造性表观遗传中的功能作用，我们将通过确定核结构修改和基因表达之间的病理联系来挑战传统的基因调控观点，这些变化是肿瘤发生的分子病因学的基础。