Subnuclear Targeting and Architectural Epigenetics in Cancer Cells

癌细胞表现出控制细胞命运和损害细胞生长控制的核结构参数的改变。我们的项目通过展示基因调控因子在染色质微环境（“亚核病灶”）中整合细胞信号，并通过与有丝分裂染色体的关联支持表观遗传机制，建立了新的生物学范式。与其他项目调查员合作。项目1现在将通过定义AML和乳腺癌细胞间期和有丝分裂期间结构相关调控机制的扰动，建立基因调控的新维度。我们的中心假设是：(1)转录因子在间期亚核靶向到基因调控位点；(2)有丝分裂染色体中转录因子与其靶基因的关联是维持正常细胞和癌细胞生物学状态的基础。因此，我们将使用IF显微镜，生化，基因组学和蛋白质组学方法(i)来表征表达易位相关t的结构表观遗传学修饰和分子病理学后果(8；21) AML-ETO融合蛋白（Aim 1）， （ii）分析乳腺癌细胞间期染色质微环境（“亚核焦点”）中由Runx2转录和空间控制的基因（Aim 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来检查Runx2介导的乳腺癌细胞中的结构表观遗传学（Aim 3）。通过研究Runx2在有丝分裂期间癌细胞间期染色质微环境（“亚核病灶”）的建立和结构表观遗传学中的功能作用，我们将通过定义核结构修饰和基因表达之间的病理联系来挑战传统的基因调控生物化学观点，这是肿瘤发生的分子病因学基础。

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.