HIJACKING OF SUPER-ENHANCERS FOR CANCER-SPECIFIC THERAPEUTICS

劫持癌症特异性治疗的超级增强剂

• 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
• 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

 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级控制，当其被破坏时可导致癌症。具有基因驱动启动子的DNA编码的增强子的染色体内环形成通过控制转录机器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不同基因组区域，可以形成跨越数十到数百千碱基的长距离染色质环。尽管增强子和典型的基因座控制区在30多年前就被发现，并通过拓扑学方法在全球范围内进行了表征，但增强子环实时形成的机制对转录领域的专家来说仍然是一个谜。在全球范围内，增强子通过与组织特异性蛋白相互作用来控制细胞命运和疾病。 启动子或关键肿瘤发病基因。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，可以为用组织特异性疗法对抗疾病提供新的进展。 该提案旨在开发一种系统，以快速诱导长距离染色体内环，用于询问和劫持增强子环机制的目的。通过将CRISPR/dCas 9基因组靶向技术与小分子诱导的邻近结合，我将能够控制染色体成环事件，从而以癌症特异性方式控制基因表达。这个系统将允许我们劫持一个活性增强子的功能复合物，并将它们物理地拴在一个受抑制的或低表达的启动子上，以控制基因表达。由于化学诱导系统允许精确的二聚化时间控制，并且核酸酶死亡（dCas 9）系统允许协调蛋白质靶向遗传区域，因此我们将能够物理地拴系在细胞的整个生命周期中相互作用的任何两个基因组区域。我们试图在癌症特异性环境中控制细胞阻滞基因的表达。此外，我们将定义在增强子形成和崩溃期间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。在这项工作的最后，我们将能够验证高阶染色质结构的controversia机制，解释为什么人类基因组的某些区域非常容易改变和易位，并验证增强子景观的重新利用癌症特异性治疗。