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（由申请人提供）：细胞命运由潜在的遗传密码、DNA 甲基化、组蛋白修饰和高级染色质结构分层控制，当这些结构被破坏时，会导致 DNA 编码的增强子在染色体内形成环。基因驱动启动子通过控制转录机制的定位来调节组织特异性基因表达。增强子是包含转录因子结合位点的独特基因组区域，并且可以形成长程。尽管增强子和典型基因座控制区域在 30 多年前就已被发现，并且通过拓扑方法进行了全球性的表征，但增强子环的实时形成机制对于该领域的专家来说仍然是一个谜。在全球范围内，增强子通过组织特异性相互作用来控制细胞的命运和疾病。因此，在分子水平上更详细地了解增强子如何形成和介导基因表达可以为通过组织特异性疗法对抗疾病提供新的进展。通过将 CRISPR/dCas9 基因组靶向技术与小分子诱导接近相结合，我将能够控制染色体环。该系统将允许我们劫持活性增强子的功能复合物，并将它们物理地连接到抑制的或低表达的启动子上以控制基因表达。诱导系统允许对二聚化进行精确的时间控制，而核酸酶死亡（dCas9）系统允许协调蛋白质靶向遗传区域，我们将能够物理束缚在细胞整个生命周期中相互作用的任何两个基因组区域。这此外，我们将定义增强子形成和崩溃过程中发生的事件的生化序列，并开发核组织中染色体循环事件的生物物理模型。这项工作，我们将能够验证高阶染色质结构的争议机制，解释为什么人类基因组的某些区域非常容易发生改变和易位，并验证增强子景观的重新利用用于癌症特异性治疗。