Role of Protein Arginine Methyltransferase 9 in Acute Myeloid Leukemia Maintenance

蛋白精氨酸甲基转移酶 9 在急性髓系白血病维持中的作用

基本信息

批准号：
10348138
负责人：
LING LI
金额：
$ 42.48万
依托单位：
BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
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：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10348138
关键词：
Acute Myelocytic Leukemia Arginine Binding Blood CD34 gene CEBPA gene Cell Cycle Cell Maintenance Cell Survival Cells Cytarabine Cytogenetics Daunorubicin Dependence Disease Elongation Factor Enzymes FLT3 gene Genetic Genetic Translation Goals Growth Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Hematologic Neoplasms Hematological Disease Hematopoietic Human Knock-in Knock-in Mouse Knockout Mice Knowledge Lead Leukemic Cell Link MCL1 gene MLL-AF9 Maintenance Malignant Neoplasms Manuscripts Mass Spectrum Analysis Mediating Messenger RNA Methylation Modeling Molecular Mus Mutagenesis Mutation Oncogenic Oncoproteins Outcome Pathogenesis Patients Peptide Elongation Factor 1 Play Protein Biosynthesis Protein-Arginine N-Methyltransferase Proteins Proteomics Relapse Risk Role Signal Transduction Small Interfering RNA Source Specimen Stat5 protein TP53 gene Testing Translations Transplantation Tyrosine Kinase Inhibitor Work acute myeloid leukemia cell base c-myc Genes cancer cell cell growth cellular transduction chemotherapy cohort conditional knockout conventional therapy dimethylarginine effective therapy interest knock-down leukemia mouse model novel overexpression patient derived xenograft model polysome profiling ribosome profiling standard care targeted treatment therapeutically effective transplant model tumor

项目摘要

Project Summary Acute myeloid leukemia (AML) is a deadly disease. Although cytogenetic and molecular risk-adapted approaches can guide treatment, overall outcomes remain poor. Currently available cell cycle-based chemotherapy or targeted therapy, such as use of tyrosine kinase inhibitors (TKIs), cannot eliminate all leukemia clones. Mounting evidence suggests that remaining cells that depend on oncoproteins for survival become a source of relapse. Many of these oncoproteins, such as Mcl-1, STAT5, and c-Myc, are short-lived, and their high levels in tumors are likely due to aberrant activation of the translation machinery, a hallmark of cancer. Thus, understanding mechanisms underlying dysregulated translation is necessary to antagonize leukemia persistence. We recently found that higher expression levels of PRMT9, the most recently defined symmetric- dimethylarginine (SDMA)-forming enzyme, are associated with decreased overall AML patient survival. Our proteomics and mutagenesis analysis revealed a novel mechanism whereby PRMT9 catalyzes methylation of translation elongation factor eEF1A1, linking PRMT9 to active translation and promoting AML maintenance. Accordingly, PRMT9 inhibition blocked AML cell survival/proliferation in an eEF1A1 methylation-dependent manner, while sparing normal hematopoietic cells. Inhibition of PRMT9/eEF1A signaling indeed decreased protein biosynthesis, reducing levels of the short-lived oncoproteins Mcl-1, c-Myc, and STAT5 and significantly extending leukemic mouse survival. Thus, we hypothesize that PRMT9 is critical for AML pathogenesis and that PRMT9-mediated eEF1A methylation promotes mRNA translation and protein synthesis to enable AML maintenance. To test this hypothesis, we will: 1) define the function of PRMT9-mediated eEF1A arginine methylation in AML pathogenesis using a MLL-AF9-related doubly-hit AML mouse model and a newly developed PRMT9 conditional knockout mouse; 2) define molecular mechanisms underlying PRMT9/eEF1A signaling in AML by testing whether eEF1A methylation alters its GTPase activity by enhancing GTP/GDP binding and assessing whether PRMT9 deficiency induces global translation changes in AML based on ribosome profiling sequencing; 3) determine combined effects of PRMT9 deletion or inhibition with daunorubicin/cytarabine or AC220 treatments on AML eradication in murine and human AML models. Our studies should reveal functional interaction between PRMT9 and eEF1A and indicate how arginine methylation of highly conserved eEF1A R166 governs translation in cancer cells. These studies will close the knowledge gap relevant to how leukemia cells acquire a survival/growth advantage through aberrant activity of the translation machinery and may show how combining targeting of PRMT9 with current treatments could represent a more effective strategy to eliminate AML cells.

项目摘要急性髓样白血病（AML）是一种致命疾病。虽然细胞遗传学和分子风险适应方法可以指导治疗，总体结果仍然很差。目前可用的基于单元的周期化学疗法或靶向疗法，例如使用酪氨酸激酶抑制剂（TKI），无法消除所有白血病克隆。越来越多的证据表明，其余依赖癌蛋白生存的细胞成为一个复发来源。这些癌蛋白中的许多，例如Mcl-1，Stat5和C-Myc，都是短暂的，它们的高肿瘤的水平可能是由于癌症标志的翻译机械的异常激活所致。因此，了解翻译失调的基础机制对于拮抗白血病是必要的持久性。我们最近发现，PRMT9的较高表达水平是最近定义的对称性二甲基精氨酸（SDMA）形成酶与AML总体患者存活降低有关。我们的蛋白质组学和诱变分析揭示了一种新的机制，PRMT9催化甲基化的甲基化翻译伸长因子EEF1A1，将PRMT9与主动翻译和促进AML维护联系起来。因此，PRMT9抑制阻断了EEF1A1甲基化依赖性的AML细胞存活/增殖方式，同时保留正常的造血细胞。 PRMT9/EEF1A信号的抑制确实减少了蛋白质生物合成，降低短寿命癌蛋白Mcl-1，C-Myc和STAT5的水平，并且显着扩展白血病小鼠的生存。因此，我们假设PRMT9对于AML发病机理至关重要，并且 PRMT9介导的EEF1A甲基化促进mRNA翻译和蛋白质合成以实现AML 维护。为了检验该假设，我们将：1）定义PRMT9介导的EEF1A精氨酸的功能使用MLL-AF9相关的双重打击AML小鼠模型和新开发的AML发病机理中的甲基化 PRMT9有条件的敲除鼠标； 2）定义PRMT9/EEF1A信号传导的分子机制通过测试EEF1A甲基化是否通过增强GTP/GDP结合和评估PRMT9缺乏是否基于核糖体分析诱导AML的全局翻译变化测序； 3）确定daunorubicin/cytarabine或 AC220关于鼠和人类AML模型中AML根除的处理。我们的研究应揭示功能 PRMT9与EEF1A之间的相互作用，并指示高度保守的EEF1A R166的精氨酸甲基化如何控制癌细胞的翻译。这些研究将缩小与白血病细胞如何相关的知识差距通过翻译机械的异常活动获得生存/成长优势，并可能显示如何将PRMT9与当前治疗的靶向相结合可以代表消除更有效的策略 AML细胞。