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.

项目概要急性髓系白血病（AML）是一种致命的疾病。尽管细胞遗传学和分子风险适应方法可以指导治疗，但总体结果仍然不佳。目前可用的基于细胞周期的化疗或靶向治疗，例如使用酪氨酸激酶抑制剂（TKI），并不能消除所有白血病克隆。越来越多的证据表明，依赖癌蛋白生存的剩余细胞变成了复发的根源。许多这些癌蛋白，例如 Mcl-1、STAT5 和 c-Myc，都是短暂的，并且它们的高活性肿瘤中的水平可能是由于翻译机器的异常激活所致，这是癌症的一个标志。因此，了解翻译失调的机制对于对抗白血病是必要的坚持。我们最近发现PRMT9（最新定义的对称- 二甲基精氨酸 (SDMA) 形成酶与 AML 患者总体生存率降低有关。我们的蛋白质组学和诱变分析揭示了 PRMT9 催化甲基化的新机制翻译延伸因子 eEF1A1，将 PRMT9 与主动翻译联系起来并促进 AML 维持。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，同时保留正常造血细胞。 PRMT9/eEF1A 信号传导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，并显着延长白血病小鼠的存活时间。因此，我们假设 PRMT9 对于 AML 发病机制至关重要，并且 PRMT9介导的eEF1A甲基化促进mRNA翻译和蛋白质合成，从而使AML成为可能维护。为了检验这个假设，我们将：1）定义 PRMT9 介导的 eEF1A 精氨酸的功能使用 MLL-AF9 相关双重打击 AML 小鼠模型和新开发的方法研究 AML 发病机制中的甲基化 PRMT9条件敲除小鼠； 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 结合来改变其 GTPase 活性来进行 AML 基于核糖体分析评估 PRMT9 缺陷是否会引起 AML 的整体翻译变化测序； 3) 确定 PRMT9 缺失或抑制与柔红霉素/阿糖胞苷的联合效应或 AC220 治疗可消除小鼠和人类 AML 模型中的 AML。我们的研究应该揭示功能性 PRMT9 和 eEF1A 之间的相互作用，并表明高度保守的 eEF1A R166 的精氨酸甲基化如何控制癌细胞中的翻译。这些研究将缩小与白血病细胞如何发生相关的知识差距通过翻译机器的异常活动获得生存/增长优势，并可能展示如何将 PRMT9 的靶向与当前治疗相结合可能是一种更有效的策略来消除 AML细胞。