Dependency of AML on CARM1 activity

项目总结/摘要 确定驱动细胞转化的遗传和表观遗传异常提供了 深入了解癌症的潜在治疗方法。然而，无数的基因变化 伴随特定类型的癌症，很难找到治疗方法 适用于大多数患有特定疾病的患者。我们已经确定了一个致癌作用， AML中的表观遗传酶“辅激活因子相关精氨酸甲基转移酶1”（CARM 1）， 以及AML细胞而非正常造血干细胞对CARM 1活性的依赖性。 CARM 1甲基化多种组蛋白和非组蛋白底物，并通过以下途径靶向其活性： 小分子抑制剂，敲低（KD）或敲除（KO）方法，触发AML细胞 分化与死亡拟议的研究代表了一种概念上的转变， AML中发现的特定突变，以开发靶向染色质修饰剂的疗法， 转录调节因子是大量AML患者所必需的。推进这一 我们提出了以下具体目标： 目标1：我们将定义CARM 1促进分化作用的分子基础 抑制AML细胞，鉴定关键的CARM 1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- 最先进的技术，包括BioID 2系统，质谱，RNA-Seq和 ChIP测序目的2：我们将确定控制CARM 1水平的机制。 AML细胞中的活性，定义了CARM 1 mRNA水平、蛋白水平和治疗后 包括磷酸化在内的转录修饰影响AML细胞中的CARM 1活性。目的 3：确定AML细胞对CARM 1抑制或KD的独特敏感性的基础。我们将 还定义了CARM 1抑制剂耐药机制，并测试了含有CARM 1抑制剂的 使用人AML细胞系和原代AML样品和NOD/SCID的联合疗法 移植了人AML细胞的小鼠。这些研究将有助于确定CARM 1的重要性。 在AML的发病机制中，并推进我们对CARM 1抑制剂如何被治疗的理解。 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.