Dependency of AML on CARM1 activity

项目摘要/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洞察。然而，无数的基因变化 与特定类型的癌症伴发的癌症使寻找治疗方法变得困难 适用于大多数患有某种疾病的患者。我们已经确定了一种致癌作用 AML中的表观遗传酶“共激活物相关精氨酸甲基转移酶1”(CARM1)， AML细胞，但不是正常的造血干细胞，对CARM1活性的依赖。 CARM1甲基化多种组蛋白和非组蛋白底物，并通过 小分子抑制剂，基因敲除(KD)或基因敲除(KO)方法，触发AML细胞 分化和死亡。拟议的研究代表了从以目标为目标的概念转变 在急性髓系白血病中发现特定突变以开发靶向染色质修饰物和 转录调控因子对大量AML患者来说是必不可少的。为了推进这一进程 新范式我们提出了以下具体目标： 目的1：我们将确定CARM1促进分化作用的分子基础 对AML细胞的抑制，确定关键的CARM1相互作用蛋白、底物和 调控髓系分化、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、质谱学、RNA-Seq和 芯片序列目标2：我们将确定控制CARM1水平的机制 AML细胞中的活性，定义CARM1 mRNA水平、蛋白水平和细胞周期后的变化 转录修饰，包括磷酸化，影响AML细胞中CARM1的活性。目标 3：确定AML细胞对CARM1抑制或KD的独特敏感性的基础。我们会 还定义了CARM1抑制剂的抗性机制，并测试了含有CARM1抑制剂的 使用人AML细胞系、原代AML样本和NOD/SCID的联合治疗 植入人急性髓系白血病细胞的小鼠。这些研究将有助于确定CARM1的重要性 在AML的发病机制中的作用，并促进我们对CARM1抑制剂如何 作为治疗癌症的有效和新颖的“表观遗传靶向”策略的一部分。

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.