HIJACKING OF SUPER-ENHANCERS FOR CANCER-SPECIFIC THERAPEUTICS

劫持癌症特异性治疗的超级增强剂

基本信息

批准号：
9050039
负责人：
Emma J Chory
金额：
$ 3.51万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9050039
关键词：
Address Automobile Driving Binding Sites Biochemical Biological Assay Biophysics Biopolymers CRISPR/Cas technology Cell Cycle Cell Cycle Arrest Cell Cycle Stage Cell Line Cell Lineage Cell Proliferation Cells Chemicals Chromatin Chromatin Loop Chromatin Structure Clustered Regularly Interspaced Short Palindromic Repeats Complex Confocal Microscopy DNA DNA Methylation Dimerization Disease Embryo Engineering Enhancers Environment Event Fibroblasts Fluorescent in Situ Hybridization Frequencies Gene Activation Gene Expression Gene Expression Regulation Genes Genetic Genetic Code Genetic Transcription Genome Genomic Instability Genomic Segment Goals Higher Order Chromatin Structure Histones Homo Human Genome Imagery Kinetics Knowledge Lead Life Locus Control Region Malignant Neoplasms Maps Measurement Measures Mechanics Mediating Methods Microscopic Modeling Molecular Motion Mus Nuclear Nucleic Acid Regulatory Sequences Pathogenesis Play Polymers Population Proteins Recruitment Activity Regulation Reporter Genes Resistance Resolution Role System Techniques Technology Therapeutic Time Tissue-Specific Gene Expression Tissues Transcription Coactivator Transcriptional Activation Domain Work atherogenesis biochemical tools biophysical model cancer cell cell type chromatin immunoprecipitation chromosome conformation capture combat epigenetic regulation gain of function histone modification inhibitor/antagonist insight interest lipid biosynthesis novel nuclease promoter public health relevance small molecule tool transcription factor tumor

项目摘要

DESCRIPTION (provided by applicant): Cellular fate is hierarchically controlled by the underlying genetic code, DNA methylation, histone modifications, and higher order chromatin structure, which when disrupted can lead to cancer. Intra- chromosomal loop-formation of DNA-encoded enhancers with gene-driving promoters regulates tissue- specific gene expression by controlling the localization of transcriptional machinery. Enhancers are distinct genomic regions containing transcription faction binding sites and can form long-range chromatin loops that span tens to hundreds of kilobases. Though enhancers and canonical locus control regions were discovered over 30 years ago and are globally characterized by topological methods, the mechanisms of enhancer-loop formation in real-time remains a mystery to experts in the field of transcription. Globally, enhancers govern cell fate and disease by interacting with tissue-specific promoters or key tumor pathogenesis genes. Thus, a more detailed understanding at the molecular level of how enhancers form and mediate gene expression could provide novel inroads into combatting diseases with tissue-specific therapies. This proposal seeks to develop a system to rapidly induce long-range intra-chromosomal loops for the purpose of interrogating and hijacking enhancer looping mechanisms. By combining the CRISPR/dCas9 genome targeting technology with small-molecule induced proximity, I will be able to control chromosomal looping events, and hence gene expression in a cancer-specific manner. This system will allow us to hijack the functional complexes of an active enhancer, and physically tether them to a repressed, or low-expressing promoter to control gene expression. Because the chemical-induced system allows precise temporal control of dimerization, and the nuclease dead (dCas9) system allows for coordinated protein targeting to genetic regions, we will be able to physically tether any two genomic regions that interact throughout the life of a cell. We seek to control the expression of cell-arresting genes in a cancer-specific environment. Further, we will both define the biochemical sequence of events that occur during enhancer formation and collapse, and develop of biophysical model of chromosomal looping events in the context of nuclear organization. At the culminating of this work, we will be able to validate the controversia mechanics of higher-order chromatin structure, explain why certain regions of the human genome are highly susceptible to alteration and translocation, and validate the repurposing of enhancer landscapes for cancer-specific therapies.

描述（由适用提供）：细胞命运在层次上受到基本遗传密码，DNA甲基化，组蛋白修饰和高阶染色质结构的控制，而在干扰后会导致癌症。 DNA编码增强子与基因驱动启动子的染色体内环形成通过控制转录机械的定位来调节组织 - 特定基因表达。增强子是含有转录因子结合位点的不同基因组区域，可以形成跨越数十千酶的远程染色质环。尽管增强子和规范的基因座控制区域是在30年前发现的，并且以拓扑方法为特征，但实时增强器 - 环形成的机制对于转录领域的专家来说仍然是一个神秘的。在全球范围内，增强剂通过与组织特异性相互作用来控制细胞命运和疾病启动子或关键肿瘤发病机理。这是在分子水平上对增强子形成和介导基因表达方式的更详细的理解，可以为用组织特异性疗法打击疾病提供新颖的侵害。该提案旨在开发一个系统，以迅速诱导远程染色体内环，以询问和劫持增强子循环机制。通过将CRISPR/DCAS9基因组靶向技术与小分子诱导的接近性相结合，我将能够以癌症特异性方式控制染色体环循环事件，从而控制基因表达。该系统将使我们能够劫持活动增强子的功能复合物，并将它们物理绑在反射或低表达的启动子中，以控制基因表达。由于化学诱导的系统允许精确地临时控制二聚化，并且核酸酶死亡（DCAS9）系统允许对通用区域进行协调的蛋白质靶向，因此我们将能够在整个细胞生命的整个生命周期内将任何两个基因组区域束缚。我们试图在癌症特异性环境中控制细胞心的基因的表达。此外，我们都将定义在增强子形成和崩溃过程中发生的事件的生化序列，并在核组织的背景下开发染色体循环事件的生物物理模型。在这项工作的最终结束时，我们将能够验证高阶染色质结构的争议力学，解释为什么人类基因组的某些区域非常容易受到改变和易位的影响，并验证重新利用增强剂景观的癌症特异性疗法。