RNA Splicing Modulators for MDS/AML

目前治疗骨髓增生异常综合征(MDS)和急性髓系白血病(AML)的方法是 不令人满意的是，在过去的一年里，我们和其他三个小组发现U2AF1和其他七个 在MDS和AML患者中，前mRNA剪接复合体的组成部分反复突变。 我们的合作者和其他人已经开发出了这种复合体的小分子调节剂。最多的 这一类中的高级化合物(称为sudemycins)是可溶的、稳定的、对肿瘤移植瘤有活性的。 在小鼠中，对正常细胞无毒，并可按克量合成。速德霉素与SF3B结合 剪接复合体，并在短暂暴露后诱导细胞系凋亡，部分是通过引起选择性剪接 MDM2是P53的负调控因子。该项目的长期目标是发展RNA剪接 调节剂作为MDS和AML的靶向治疗药物，我们的初步数据显示敏感性增强 表达突变型U2AF1的细胞对苏德霉素的抗性。我们假设RNA剪接机制中的突变 通过失调选择性剪接产物的表达来增强对剪接调节剂的敏感性 参与MDS/AML的发病。我们将在体外和体内使用定义良好的系统来验证这一假设 活着。在特定目标1中，具有或不具有剪接基因突变的原代人AML成髓细胞，基于 对这些样本中所有编码基因的完整测序将用于评估 Sudemycins(和其他剪接调节剂)在体外对生长、活力、凋亡和细胞周期动力学的影响 免疫缺陷异种移植后体内嵌合体、细胞凋亡和三倍体分化 老鼠。这将决定剪接基因突变是否可以作为对这些药物敏感性的生物标记物。 在特定的目标2中，我们将发现剪接调节子和剪接调节剂引起的转录组的变化 开发灵敏和定量的分析方法，以评估这些制剂的靶向效应。这 该项目通过识别可预测的遗传生物标记物来解决孢子翻译终点问题 对剪接调节剂的反应和开发用于药效学的分析方法 剪接调节剂临床试验中的监测： 相关性(请参阅说明)： 目前的化疗方法治愈了少数骨髓增生异常综合征(MDS)患者和 急性髓系白血病(AML)。我们和其他人最近发现，RNA剪接是一种关键的 代谢途径在许多患者中发生了变化，调节这一途径的化合物是 可用。该项目将测试这种新型化合物在人类细胞中的活性，并开发出 在MDS或AML患者中启动这些药物的临床试验所需的分析和临床前数据。

Current therapies for myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) are unsatisfactory, During the past year, we and three other groups discovered that U2AF1 and seven other components of the pre-mRNA splicing complex are recurrently mutated in patients with MDS and AML. Small molecule modulators of this complex have,been developed by our collaborators and others. The most advanced compounds in this class (called sudemycins) are soluble, stable, active against tumor xenografts in mice, non-toxic to normal cells, and can be synthesized in gram quantities. Sudemycins bind the SF3b splicing complex and induce apoptosis in cell lines after brief exposure, in part by causing alternative splicing of MDM2, a negative regulator of P53. The long-term goal of this proiect is to develop RNA splicing modulators as targeted therapv for MDS and AML, Our preliminary data demonstrate enhanced sensitivity of cells expressing mutant U2AF1 to sudemycin. We hypothesize that mutations in RNA splicing machinery confer sensitivity to splicing modulators by dysreguiating expression of alternatively spliced products that contribute to MDS/AML pathogenesis. We will test this hypothesis using well-defined systems in vitro and in vivo. In Specific Aim 1, primary human AML myeloblasts with or without splicing gene mutations, based on completed sequencing of all coding genes in these samples, will be used to assess the impact of sudemycins (and other splicing modulators) on growth, viability, apoptosis, and cell cycle kinetics in vitro and chimerism, apoptosis, and trilineage differentiation in vivo after xenotransplantation into immune deficient mice. This will determine whether splicing gene mutations serve as biomarkers of sensitivity to these drugs. In Specific Aim 2, we will discover alterations in the transcriptome induced by splicing modulators and develop sensitive and quantitative assays for assessment of the on-target effects of these agents. This project addresses SPORE translational endpoints by identifying genetic biomarkers that are predictive of response to splicing modulators and by developing assays that will be used for pharmacodynamic monitoring in clinical trials of splicing modulators: RELEVANCE (See instructions): Current chemotherapy treatments cure a minority of patients with myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). We and others have recently discovered that RNA splicing, a critical metabolic pathway, is altered in many of these patients and compounds that modulate this pathway are available. This project will test the activity of this novel class of compounds in human cells and develop assays and preclinical data needed to initiate clinical trials of these agents in patients with MDS or AML.