Defining the Roles of Polycomb Repressive Complex 2 (PRC2) Subcomplexes in H3 K27M Gliomas

定义 Polycomb 抑制复合物 2 (PRC2) 子复合物在 H3 K27M 胶质瘤中的作用

• 批准号: 10389924
• 负责人: Tyler J Reich
• 金额: $ 4.11万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10389924
• 关键词: Ablation; Achievement; Address; Affinity; Applications Grants; Binding; Biochemical; Biochemistry; Biological Assay; Biology; Brain Neoplasms; CRISPR/Cas technology; Cancer Biology; Cell Maintenance; Cell Proliferation; Cells; Child; Childhood; Childhood Brain Neoplasm; Childhood Malignant Brain Tumor; Chromatin; Chromatin Structure; Clinical; Clinical Trials; Collaborations; Complex; CpG Islands; DNA; Data; Development; Diffuse; Diffuse intrinsic pontine glioma; Disease; Ensure; Environment; Event; Fatal Outcome; Foundations; Future; Gene Expression; Gene Silencing; Genes; Genome; Genomics; Glioma; Gliomagenesis; Higher Order Chromatin Structure; Histone H3; Histones; Human; In Vitro; Kinetics; Life; Lysine; Malignant Neoplasms; Measures; Mediating; Mentors; Methionine; Methylation; Methyltransferase; Modeling; Molecular; Multienzyme Complexes; Mutate; Mutation; Outcome; Pathogenesis; Patients; Physicians; Polycomb; Post-Translational Protein Processing; Predisposition; Prognosis; Proteomics; Recurrence; Regulation; Reporting; Research; Research Personnel; Residual state; Resistance; Resources; Role; Scientist; Series; Site; Testing; Therapeutic; Tissues; Training; Tumor Suppressor Genes; Tumor Suppressor Proteins; Universities; Variant; Wisconsin; Work; anti-cancer; base; career; cell growth; cell type; defined contribution; design; experimental study; genome-wide; glioma cell line; human disease; improved; inhibitor; innovation; insight; loss of function mutation; mortality; mouse model; novel therapeutics; oncohistone; preclinical efficacy; prevent; promoter; recruit; therapeutic development; therapeutic target; tumor; tumorigenesis; virtual

PROJECT ABSTRACT/SUMMARY Cell type-specific gene expression, the basis for complex multicellular life, is achieved through packaging a common, invariant genome into higher-order chromatin structures with myriad levels of regulation. Covalent post- translational modifications (PTMs) at specific histone residues are major points of regulation that are dynamically modified throughout development to ensure proper tissue lineage specification, cell fate transitions, and maintenance of cell identity. Mutations in histone modifiers, as well as the histone substrates themselves, underlie a variety of human diseases including cancer. Recurrent lysine-to-methionine mutations at residue 27 of histone H3 (H3 K27M) are found in >80% of diffuse intrinsic pontine gliomas (DIPG), a high-grade pediatric brain cancer with dismal prognosis. The H3 K27M oncohistone drives gliomagenesis in part through its inhibition of polycomb repressive complex 2 (PRC2), the methyltransferase complex responsible for methylating H3K27, resulting in genome-wide depletion of the repressive mark H3K27me3. This proposal endeavors to address a long-observed yet poorly characterized aspect of H3K27M-driven PRC2 dysregulation: residual PRC2 activity at focal sites of H3K27me3. Leveraging biochemical, genomic, and cell- based approaches, the experiments described in this study seek to define the PRC2 subunits necessary for targeting to, and catalytic activity at, sites of PRC2 recruitment in DIPG cells. These results stand to significantly deepen our understanding of H3K27M-dysregulated chromatin and may reveal exploitable vulnerabilities for H3 K27M diffuse gliomas. The training plan outlined in this proposal will be performed at the University of Wisconsin-Madison within an excellent institutional research environment boasting cutting-edge resources, facilities, and dynamic opportunities for collaboration. Execution of this training plan, under the guidance of expert mentors and collaborators in fields of cancer biology, chromatin biochemistry, proteomics, and genomics, will provide an invaluable foundation meant to launch a successful career as a physician-scientist cancer researcher.

项目摘要/总结 细胞类型特异性基因表达是复杂多细胞生命的基础，是通过包装 将常见的、不变的基因组转化为具有无数调控水平的高级染色质结构。共价后 特定组蛋白残基的翻译修饰 (PTM) 是动态调节的主要点 在整个发育过程中进行修改，以确保适当的组织谱系规范、细胞命运转变，以及 维持细胞身份。组蛋白修饰物以及组蛋白底物本身的突变， 是包括癌症在内的多种人类疾病的根源。残基 27 处反复发生赖氨酸至蛋氨酸突变 组蛋白 H3 (H3 K27M) 存在于 >80% 的弥漫性内质性脑桥胶质瘤 (DIPG) 中，这是一种高级儿科肿瘤 脑癌预后不佳。 H3 K27M 癌组蛋白部分通过其抑制作用驱动神经胶质瘤发生 多梳抑制复合物 2 (PRC2)，负责甲基化 H3K27 的甲基转移酶复合物， 导致全基因组范围内抑制性标记 H3K27me3 的缺失。 该提案致力于解决 H3K27M 驱动的 PRC2 的一个长期观察但特征不明的方面 失调：H3K27me3 焦点部位残留 PRC2 活性。利用生物化学、基因组和细胞 基于方法，本研究中描述的实验旨在定义 PRC2 亚基 DIPG 细胞中 PRC2 募集位点的靶向及其催化活性。这些结果显着 加深我们对 H3K27M 染色质失调的理解，并可能揭示 H3 可利用的漏洞 K27M 弥漫性胶质瘤。 本提案中概述的培训计划将在威斯康星大学麦迪逊分校执行 优良的机构研究环境，拥有尖端的资源、设施和活力 合作的机会。本培训计划的执行，在专家导师的指导下进行 癌症生物学、染色质生物化学、蛋白质组学和基因组学领域的合作者将提供 为作为一名医师科学家癌症研究人员开启成功的职业生涯奠定了宝贵的基础。