Role of Protein Methylation in Cell Mitosis and Glioblastoma

蛋白质甲基化在细胞有丝分裂和胶质母细胞瘤中的作用

基本信息

批准号：
10322748
负责人：
Shi-Yuan Cheng
金额：
$ 42.96万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-15 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10322748
关键词：
Address Affect Arginine Biological Cell Shape Cell division Cell physiology Cells Chromatin Chromosome Condensation Chromosomes Clinical Combined Modality Therapy DNA Damage Enzymes Epigenetic Process Event Gene Expression Genetic Glioblastoma Goals Growth Guanine Nucleotide Exchange Factors Guanosine Triphosphate Phosphohydrolases Histones In Vitro Knock-out Knowledge Learning Malignant - descriptor Malignant Neoplasms Messenger RNA Methylation Methyltransferase Mitosis Mitotic Molecular Nuclear Organelles Patient-Focused Outcomes Patients Phenotype Phosphorylation Phosphotransferases Physical condensation Play Post-Translational Protein Processing Process Prognostic Marker Property Protein Analysis Protein Family Protein Inhibition Protein Methylation Protein-Arginine N-Methyltransferase Protein-Serine-Threonine Kinases Proteins RNA Splicing Radiation therapy Regulation Role Signal Pathway Signal Transduction Substrate Interaction Testing Threonine Transferase Tumor Biology Tumorigenicity antitumor effect casein kinase II clinical application enzyme substrate experimental study improved in silico in vivo innovation non-histone protein novel pre-clinical research protein arginine methyltransferase 2 protein function radiation response recruit response small molecule standard care standard of care temozolomide therapeutic target treatment response treatment strategy tumor tumor growth

项目摘要

Methylation on histone and non-histone proteins is a common and important post-translational modification (PTM) that controls protein functions and activities in modulating cancer tumorigenicity and responses to therapies. In this project, we will investigate the mechanism for protein arginine (R) methylation regulation of GBM tumorigenicity by elucidating the role of protein arginine methyltransferase 6 (PRMT6) on cell mitosis through specific arginine methylation of regulator of chromosome condensation 1 (RCC1), thereby regulating GBM phenotype and responses to therapies. We provide novel evidence that PRMT6 regulates RCC1 activity through methylation of RCC1 at R214, the arginine residue required for RCC1 activation of Ran GTPase through association with histones in chromosome, thereby affecting cell mitotic process, GBM tumorigenicity and responses to therapies. We demonstrate that casein kinase 2 (CK2), a ubiquitously expressed and constitutively active serine/threonine kinase that is important in cancers stimulates PRMT6 methyltransferase activity by phosphorylating threonine 21 of PRMT6 protein. Additionally, genetic depletions or small molecule targeting of PRMT6, RCC1 or CK2 inhibited GBM tumorigenicity in vitro and in vivo, and enhanced anti-tumor effects by radiation therapy (RT). These strong scientific premises provide the basis for the overarching hypothesis of this proposal: CK2-activated PRMT6 induces asymmetric dimethylation (aDMA) of RCC1, thereby regulating cell mitosis, GBM tumorigenicity as well as GBM responses to RT. We will address this hypothesis in the context of three specific aims: 1) Determine how PRMT6-induced R214 aDMA of RCC1 affects cell mitotic process as well as GBM tumor biologic properties in vitro and in vivo; 2) Define mechanisms by which CK2 regulates PRMT6 activity, and the relationships between CK2-associated phosphorylation of PRMT6, RCC1 function, cell mitosis, GBM tumor phenotypes, and association with GBM patient outcome; and 3) Determine whether PRMT6 and CK2 directed combination therapy is more effective than corresponding monotherapies, and whether such combination therapy enhances the anti-tumor activity of RT and temozolomide (TMZ) treatment. At the completion of this project, we will learn how PRMT6-catayzed aDMA of RCC1 shapes of cell mitosis, GBM phenotype and response to therapy. Our proposed studies will be the first to address the role of PRMT activity on non-histone aDMA of RCC1 in cell mitosis, GBM tumor biology and responses to therapies. This knowledge, in turn, will provide clear indication of the potential benefit of including combination of inhibition of PRMT6 and CK2 with RT and TMZ as part of treatment strategy for patient with GBM.

组蛋白和非组蛋白的甲基化是一种常见且重要的翻译后过程修饰（PTM），控制蛋白质功能和活性，调节癌症致瘤性和对治疗的反应。在这个项目中，我们将研究蛋白质精氨酸（R）甲基化的机制通过阐明蛋白精氨酸甲基转移酶 6 (PRMT6) 对细胞的作用来调节 GBM 致瘤性通过染色体浓缩调节因子 1 (RCC1) 的特异性精氨酸甲基化来进行有丝分裂，从而调节 GBM 表型和对治疗的反应。我们提供了 PRMT6 调节 RCC1 的新证据通过 RCC1 R214 甲基化来发挥活性，R214 是 RCC1 激活 Ran GTPase 所需的精氨酸残基通过与染色体中的组蛋白结合，从而影响细胞有丝分裂过程、GBM致瘤性和对治疗的反应。我们证明酪蛋白激酶 2 (CK2) 是一种普遍表达且组成型的在癌症中很重要的活性丝氨酸/苏氨酸激酶通过刺激 PRMT6 甲基转移酶活性磷酸化 PRMT6 蛋白的苏氨酸 21。此外，基因缺失或小分子靶向 PRMT6、RCC1或CK2在体外和体内抑制GBM致瘤性，并通过以下方式增强抗肿瘤作用：放射治疗（RT）。这些强有力的科学前提为这一总体假设提供了基础提议：CK2激活的PRMT6诱导RCC1的不对称二甲基化（aDMA），从而调节细胞有丝分裂、GBM 致瘤性以及 GBM 对 RT 的反应。我们将在以下背景下讨论这个假设：三个具体目标：1) 确定 PRMT6 诱导的 RCC1 R214 aDMA 如何影响细胞有丝分裂过程 GBM 肿瘤的体外和体内生物学特性； 2) 定义CK2调节PRMT6的机制活性，以及 CK2 相关的 PRMT6 磷酸化、RCC1 功能、细胞有丝分裂、 GBM 肿瘤表型以及与 GBM 患者预后的关联； 3) 确定 PRMT6 和 CK2定向联合疗法比相应的单一疗法更有效，以及是否这样联合治疗增强了 RT 和替莫唑胺 (TMZ) 治疗的抗肿瘤活性。在完成这个项目后，我们将了解PRMT6如何催化RCC1形状的细胞有丝分裂、GBM的aDMA 表型和对治疗的反应。我们提出的研究将是第一个探讨 PRMT 活动的作用的研究 RCC1 的非组蛋白 aDMA 在细胞有丝分裂、GBM 肿瘤生物学和治疗反应中的作用。这些知识，反过来，将清楚地表明包括 PRMT6 抑制和 CK2 联合 RT 和 TMZ 作为 GBM 患者治疗策略的一部分。