Project 2: Elucidating Mechanisms of Chromatin Dysregulation by Oncohistones

项目 2：阐明肿瘤组蛋白染色质失调的机制

基本信息

批准号：
10024844
负责人：
CHARLES DAVID ALLIS
金额：
$ 27.88万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-09 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10024844
关键词：
ATAC-seq Affect Alleles Allografting Animal Model Bar Codes Biological Assay Biology Cancer Biology Cancer cell line Cell Differentiation process Cell Fate Control Cell Line Cell physiology Cells Cellular Structures ChIP-seq Characteristics Charge Chemicals Childhood Glioma Chondroblastoma Chromatin Chromatin Structure Collaborations Complementary DNA Cues DNA DNA Damage DNA Methylation DNA Modification Process DNA Repair Data Defect Development Epigenetic Process Frequencies Gene Expression Genetic Genetic Screening Genetic Transcription Genome Goals Grant Growth Histone H3 Histones Human Knock-in Laboratories Lead Libraries Light Link Lysine Malignant Childhood Neoplasm Malignant Neoplasms Mass Spectrum Analysis Mediating Mesenchymal Stem Cells Methionine Missense Mutation Modeling Modification Molecular Mus Mutate Mutation N-terminal Nucleosomes Oncogenic Pathologic Pathway interactions Patients Pharmacology Phenotype Play Positioning Attribute Post-Translational Protein Processing Pre-Clinical Model Process Property Proteins Reader Recurrence Research Research Personnel Role Site Somatic Mutation Structure Tail Technology Testing Therapeutic Translating Transplantation Tumor Subtype Undifferentiated Variant Work Writing Xenograft procedure cancer cell cancer type cell type cellular transduction design driver mutation epigenome gain of function genome-wide histone methyltransferase histone modification in vivo insight interdisciplinary approach mouse model multidisciplinary novel prevent programs response sarcoma self-renewal small molecule small molecule inhibitor success transcriptome sequencing transcriptomics translational approach tumor tumorigenesis tumorigenic

项目摘要

PROJECT SUMMARY (ALLIS) Genome-wide sequencing technologies have allowed an unprecedented discovery of somatic mutations in chromatin and epigenetic modifiers in human cancers, providing mechanistic links between cancer epigenomes and genetic alterations. The collective number of oncogenic mutations in epigenetic regulators has led to the emerging view of “driver mutations” underlying cancer epigenomes. Nowhere is this better illustrated than with the now classical findings of high-frequency (50-95%) missense mutations in core histones, such as histone H3 lysine 27 to methionine (H3K27M) mutation in pediatric gliomas, and H3 lysine 36 to methionine (H3K36M) mutations in chondroblastomas and undifferentiated sarcomas. During the prior grant period, we have shown that these mutations directly prevent the ‘writing’ of some critical regulatory histone post-translational modifications (PTMs) to promote oncogenesis through altered chromatin organization, transcription, and in some cases cell fate and differentiation. More recently, we have extended our understanding of the landscape of histone mutations in cancers. We characterized an unexpectedly broad landscape of novel oncohistone mutations that occur in roughly 4% of all cancers. These mutations are found not only in the H3 N-terminal tail, which is the site of classical oncohistones, but also in the globular domain and in all four core histones. Our preliminary data suggest that a least a subset of these mutations affect one or more properties of chromatin and chromatin-dependent processes including nucleosome stability, histone PTMs, and cellular differentiation. We therefore hypothesize that novel oncohistone mutations will impact the landscape of histone PTMs and chromatin organization in a context dependent manner, leading to dysregulation of gene expression and effects on cell fate and tumorigenesis. The goal of this work is to rigorously test these hypotheses for a comprehensive set of cancer-associated histone mutations using a multidisciplinary approach that include genetics (barcoded oncohistone libraries, mouse models, barcoded-cell lines), epigenetics (ChIP-seq, ATAC-seq, DNA-methylation profiling), transcriptomics (RNA-seq), and chemical biology (“designer chromatin”, small molecule inhibitors). Specifically, we will 1) define molecular mechanisms by which novel oncohistones act and their impact on chromatin and gene expression; 2) determine how these molecular changes translate into phenotypes using cellular differentiation and tumor allograft models, and explore pharmacologic strategies to rescue differentiation blockade; and 3) extend our studies into animal models and diverse cellular contexts to test the roles of novel oncohistones in tumorigenesis and development. Together, these approaches will shed light on the function of newly discovered oncohistones and provide important insight into the role of histones and chromatin structure in tumorigenesis. Our findings are expected to pave new avenues towards intervening pharmacologically the aberrant epigenetic pathways for cancer therapeutics. To facilitate the success of this proposal, a world-class team of investigators, experts in cancer, chromatin and chemical biology, have been assembled.

项目摘要（Allis）全基因组的测序技术允许在史无前例的中发现体细胞突变人类癌症中的染色质和表观遗传学修饰剂，提供癌症基因组之间的机械联系和遗传改变。表观遗传调节剂中的集体致癌突变导致新兴的“驾驶员突变”潜在癌症表观性观测的观点。没有比这更好的说明了现在的核心组蛋白中高频（50-95％）错义突变的古典发现，例如组蛋白H3 赖氨酸27至小儿神经胶质瘤中的方法（H3K27M）突变，H3赖氨酸36至Metagine（H3K36M）软骨细胞瘤和未分化的肉瘤的突变。在以前的赠款期间，我们已经表明这些突变直接阻止了某些关键调节性的Hisstone翻译后的“写作” 修改（PTMS）通过改变染色质组织，转录和某些改变病例命运和分化。最近，我们扩展了对景观的理解癌症中的Hisstone突变。我们描述了一个令人意外的宽阔的酒精酮景观大约4％的所有癌症发生的突变。这些突变不仅在H3 N末端尾部，还发现这是古典酒精酮的所在地，也是全球领域和所有四个核心Hisons的地点。我们的初步数据表明，至少这些突变的一个子集会影响染色质的一个或多个特性染色质依赖性过程，包括核小体稳定性，组蛋白PTM和细胞分化。我们因此，假设新颖的酒精酮突变会影响Hisstone PTM和染色质的景观以上下文依赖性方式组织，导致基因表达和对细胞命运的影响失调和肿瘤发生。这项工作的目的是严格检验这些假设的全面集合使用包括遗传学（条形码）的多学科方法与癌症相关的组蛋白突变关于酒精酮库，小鼠模型，条形码细胞线），表观遗传学（chip-seq，atac-seq，DNA-甲基化分析），转录组学（RNA-SEQ）和化学生物学（“设计器染色质”，小分子抑制剂）。具体而言，我们将1）定义新颖的酒精作用及其对它们对的分子机制染色质和基因表达； 2）确定这些分子变化如何使用细胞分化和肿瘤同种异体移植模型，并探索挽救分化的药物策略 blocade; 3）将我们的研究扩展到动物模型和各种细胞环境中，以测试新颖的作用关于肿瘤发生和发育中的酒精酮。这些方法在一起将阐明在酒精酮上新发现，并为组蛋白和染色质结构在肿瘤发生。我们的发现有望将新的途径铺平到介入药物的情况下癌症治疗的异常表观遗传途径。为了促进该提议的成功，世界一流的研究人员，癌症，染色质和化学生物学专家团队已经组装。