Dissecting the molecular mechanisms of PRC2 dysregulation in cancer

剖析癌症中 PRC2 失调的分子机制

• 批准号: 10063688
• 负责人: Agata Ewa Patriotis
• 金额: $ 4.7万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063688
• 关键词: Address; Allosteric Regulation; Architecture; Award; B-Cell Lymphomas; Binding; Biochemistry; Cancer Biology; Cancer Model; Catalysis; Cell Communication; Cell physiology; Cells; Chromatin; Chromatin Remodeling Factor; Clonal Evolution; Clone Cells; Complex; Coupled; Data; Development; Drug resistance; EZH2 gene; Ectoderm; Embryo; Epigenetic Process; Focus Groups; Gene Expression; Gene Mutation; Genetic Screening; Genomics; Goals; Heterochromatin; Heterogeneity; Histone H3; Histones; Human; In Vitro; Lymphoma; Lysine; Maintenance; Malignant - descriptor; Malignant Neoplasms; Methylation; Microscopy; Modeling; Modification; Molecular; Molecular Biology; Mutate; Mutation; N-terminal; Neoplasms; Neoplastic Processes; Neurofibrosarcoma; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Polycomb; Post-Translational Protein Processing; Process; Processed Genes; Proliferating; Radiation therapy; Regulation; Research; Resistance; Role; Somatic Mutation; System; Technical Expertise; Techniques; Technology; Therapeutic; Therapeutic Intervention; Work; cancer cell; cancer prevention; cancer therapy; chemotherapy; drug development; epigenetic regulation; epigenomics; experience; gain of function; gain of function mutation; genome-wide; genotoxicity; in vivo; insight; large cell Diffuse non-Hodgkin' s lymphoma; loss of function; loss of function mutation; malignant state; mutant; novel; novel strategies; preclinical development; programs; reconstitution; shift work; transcriptomics; tumor

Project Summary/Abstract The chromatin landscape governs basic cellular functions that are altered in cancer, including genomic architecture, gene expression, and developmental pathways. Interestingly, epigenetic dysregulation of chromatin is an emerging hallmark of cancer. These epigenetic changes in turn render cancer cells highly reliant on the chromatin machinery to maintain their malignant state, thus creating opportunities for therapeutic intervention by targeting chromatin modifiers. Driven by the desire to understand the basic mechanistic underpinnings of epigenetic regulation, it is my goal to address pressing questions in molecular biology and contribute to the advancement of cancer prevention and treatment. Histone post-translational modifications (PTMs) are central regulators of chromatin processes, and genes encoding chromatin factors are highly mutated in a range of cancers. This project seeks to understand the role of the Polycomb Repressive Complex 2 (PRC2) in cancer development. PRC2 is a major epigenetic machinery responsible for the maintenance of heterochromatin and catalysis of histone H3 lysine 27 methylation. The F99 phase of this proposal is focused on investigating the regulation of PRC2 enzymatic activity by the highly conserved SANT1-like binding (SBD) domain of its EZH2 subunit. Despite the broad understanding of PRC2 function and regulation, the molecular role of the N-terminal SBD of EZH2 is unknown. The preliminary data reveals novel mechanistic insight about this domain in the catalysis of H3K27 methylation. Surprisingly, partial deletion of the SBD domain in EZH2 (SBD-EZH2) leads to a global loss of repressive H3K27me2 and H3K27me3, phenocopying the complete loss of EZH2 at the epigenomic level. In the remainder of the dissertation work, my main research efforts will be directed toward delineating the regulatory significance of the EZH2-SBD domain in the allosteric activation of PRC2 enzymatic activity, as well as determining a potential inhibitory mechanism for lymphoma patients harboring EZH2 gain-of-function mutations. The K00 phase of this project will be focused on studying the role of PRC2 loss in the development of the highly aggressive malignant peripheral nerve sheath tumors (MPNSTs). Interestingly, the loss of PRC2 components is involved in the malignant formation of sporadic and radiotherapy-associated MPNSTs. Thus, to further understand the molecular mechanisms of these tumors, I plan to expand my technical expertise to include high-throughput genetic screening, single-cell epigenomic and transcriptomic techniques, computational approaches, development of preclinical cancer models, and sequencing analysis of human tumor data. These new approaches, coupled with my already strong background in molecular biology, microscopy, and biochemistry, will allow me to address the most pressing and challenging issues in epigenetic regulation and cancer biology today. This award will allow to pursue the above questions and and gain experience in order to become a leader of my own cancer-focused group and a leader in the field of epigenetics.

项目总结/摘要 染色质景观控制着癌症中改变的基本细胞功能，包括 基因组结构、基因表达和发育途径。有趣的是，表观遗传失调 染色质是癌症的一个新的标志。这些表观遗传变化反过来使癌细胞高度依赖于 在染色质机器上维持其恶性状态，从而为治疗创造机会。 通过靶向染色质修饰剂进行干预。受理解基本机械原理的欲望驱使， 作为表观遗传调控的基础，我的目标是解决分子生物学中的紧迫问题， 有助于癌症预防和治疗的进步。组蛋白翻译后修饰 （PTM）是染色质过程的中心调节因子，编码染色质因子的基因高度突变 在一系列癌症中。该项目旨在了解Polycomb Repressive Complex 2（PRC 2）的作用。 在癌症发展中的作用PRC 2是一个主要的表观遗传机制，负责维持 异染色质和组蛋白H3赖氨酸27甲基化催化。 该提案的F99阶段集中于研究PRC 2酶活性的调节， 其EZH 2亚基的高度保守的SANT 1样结合（SBD）结构域。尽管广泛的理解 PRC 2功能和调节的研究中，EZH 2的N-末端SBD的分子作用是未知的。的 初步数据揭示了关于该结构域在H3 K27甲基化催化中的新的机理见解。 令人惊讶的是，EZH 2中SBD结构域的部分缺失（SBD-EZH 2）导致抑制性蛋白的全面丧失 H3 K27 me 2和H3 K27 me 3，表型复制EZH 2在表观基因组水平上的完全丢失。在 在论文的其余部分，我的主要研究工作将是针对划定监管 EZH 2-SBD结构域在PRC 2酶活性的变构活化中的意义，以及 确定携带EZH 2功能获得性突变的淋巴瘤患者的潜在抑制机制。 该项目的K 00阶段将重点研究PRC 2损失在以下发展中的作用： 高度侵袭性恶性外周神经鞘瘤（MPNSTs）。有趣的是，PRC 2的损失 成分参与散发性和放射治疗相关MPNST的恶性形成。从而 为了进一步了解这些肿瘤的分子机制，我计划扩大我的技术专长，包括 高通量遗传筛选，单细胞表观基因组学和转录组学技术，计算 方法、临床前癌症模型的开发和人类肿瘤数据的测序分析。这些 新的方法，再加上我在分子生物学，显微镜， 生物化学，将使我能够解决表观遗传调控中最紧迫和最具挑战性的问题， 今天的癌症生物学这个奖项将允许追求上述问题和获得经验，以 成为我自己的癌症小组的领导者，也成为表观遗传学领域的领导者。