Dependency of AML on CARM1 activity

AML 对 CARM1 活性的依赖性

• 批准号: 10202528
• 负责人: Stephen D. Nimer
• 金额: $ 39.01万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10202528
• 关键词: AML1-ETO fusion protein; Affect; Arginine; Blood Cells; Cancer Cell Growth; Cancer Patient; Cell Death; Cell Differentiation process; Cell Line; Cell Survival; Cells; ChIP-seq; Chromatin Remodeling Factor; Co-Immunoprecipitations; Combined Modality Therapy; Dependence; Development; Disease; Enzymes; Epigenetic Process; Genes; Genetic; Genetic Transcription; Goals; Hematopoietic; Hematopoietic stem cells; Histones; Human; Human Cell Line; JAK2 gene; Knock-out; Malignant Neoplasms; Mass Spectrum Analysis; Messenger RNA; Modeling; Modification; Molecular; Mutation; Myelogenous; NOD/SCID mouse; Nuclear; Oncogenes; Oncogenic; Pathogenesis; Pathway interactions; Patients; Phosphorylation; Pre-Clinical Model; Protein-Arginine N-Methyltransferase; Proteins; RUNX1 gene; Regulation; Role; Sampling; Signal Transduction; Specificity; Survival Rate; System; Techniques; Testing; Therapeutic; Toxic effect; Tyrosine; Up-Regulation; arginine methyltransferase; base; cancer cell; cancer type; cell growth; cell type; coactivator-associated arginine methyltransferase 1; effective therapy; experimental study; improved; inhibitor/antagonist; insight; knock-down; leukemia; metaplastic cell transformation; mouse model; non-histone protein; novel; protein protein interaction; resistance mechanism; small molecule inhibitor; targeted cancer therapy; targeted treatment; therapeutically effective; therapy development; transcription factor; transcriptome sequencing

Project Summary/ Abstract Defining the genetic and epigenetic abnormalities that drive cellular transformation has provided insights into potential therapeutic approaches to cancer. However, the myriad of genetic changes that accompany specific types of cancer makes it difficult to find therapeutic approaches applicable to the bulk of patients with a given disease. We have identified an oncogenic role for the epigenetic enzyme “co-activator associated arginine methyltransferase 1” (CARM1) in AML, and a dependency of AML cells, but not normal hematopoietic stem cells, on CARM1 activity. CARM1 methylates a variety of histone and non-histone substrates and targeting its activity, via small molecule inhibitors, knockdown (KD) or knockout (KO) approaches, triggers AML cell differentiation and death. The proposed studies represent a conceptual shift from targeting specific mutations found in AML to developing therapies that target chromatin modifiers and transcriptional regulators that are essential to large numbers of AML patients. To advance this new paradigm we have proposed the following specific aims: Aim 1: We will define the molecular basis for the differentiation-promoting effects of CARM1 inhibition on AML cells, identifying the critical CARM1-interacting proteins, substrates and target genes that regulate myeloid differentiation, proliferation and survival. We will use state-of- the-art techniques including the BioID2 system, mass-spectrometry, RNA-Seq and ChIP-Seq. Aim 2: We will determine the mechanisms that control the level of CARM1 activity in AML cells, defining how changes in CARM1 mRNA levels, protein levels, and post- transcriptional modifications, including phosphorylation, affect CARM1 activity in AML cells. Aim 3: Determine the basis for the unique sensitivity of AML cells to CARM1 inhibition or KD. We will also define CARM1 inhibitor resistance mechanisms, and test CARM1 inhibitor-containing combination therapies, using human AML cell lines and primary AML samples and NOD/SCID mice engrafted with human AML cells. These studies will help establish the importance of CARM1 in the pathogenesis of AML, and advance our understanding of how CARM1 inhibitors can be used as part of an effective and novel “epigenetic-targeted” strategy for treating cancer.

项目概要/摘要 定义驱动细胞转化的遗传和表观遗传异常提供了 对癌症潜在治疗方法的见解。然而，无数的基因变化 伴随特定类型的癌症使得很难找到治疗方法 适用于大多数患有特定疾病的患者。我们已经确定了致癌作用 AML 中的表观遗传酶“辅激活剂相关精氨酸甲基转移酶 1”(CARM1)， AML 细胞（而非正常造血干细胞）对 CARM1 活性的依赖性。 CARM1 甲基化多种组蛋白和非组蛋白底物并靶向其活性，通过 小分子抑制剂、敲低 (KD) 或敲除 (KO) 方法，触发 AML 细胞 分化和死亡。拟议的研究代表了从瞄准目标的概念转变 AML 中发现的特定突变可用于开发针对染色质修饰剂的疗法 对大量 AML 患者至关重要的转录调节因子。为了推进这个 新范式我们提出了以下具体目标： 目标1：我们将定义CARM1促进分化作用的分子基础 对 AML 细胞的抑制，识别关键的 CARM1 相互作用蛋白、底物和 调节骨髓分化、增殖和存活的靶基因。我们将使用状态- 最先进的技术，包括 BioID2 系统、质谱、RNA-Seq 和 ChIP 测序。目标2：我们将确定控制CARM1水平的机制 AML 细胞中的活性，定义了 CARM1 mRNA 水平、蛋白质水平和后处理如何变化 转录修饰（包括磷酸化）会影响 AML 细胞中的 CARM1 活性。目的 3：确定 AML 细胞对 CARM1 抑制或 KD 的独特敏感性的基础。我们将 还定义了CARM1抑制剂耐药机制，并测试了含有CARM1抑制剂的药物 联合疗法，使用人类 AML 细胞系和原发性 AML 样本以及 NOD/SCID 移植了人类 AML 细胞的小鼠。这些研究将有助于确定 CARM1 的重要性 AML 的发病机制，并加深我们对 CARM1 抑制剂如何发挥作用的理解 用作治疗癌症的有效且新颖的“表观遗传靶向”策略的一部分。