RNA Splicing Modulators for MDS/AML

骨髓增生异常综合征（MDS）和急性髓性白血病（AML）的当前疗法是 在过去的一年里，我们和其他三个小组发现，U2 AF 1和其他七个小组， 前体mRNA剪接复合物的组分在MDS和AML患者中反复突变。 我们的合作者和其他人已经开发了这种复合物的小分子调节剂。最 这类高级化合物（称为sudemycins）是可溶的、稳定的、对肿瘤异种移植物有活性的 在小鼠中，对正常细胞无毒，并且可以克量合成。苏地霉素结合SF 3b 剪接复合物，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 MDM 2是P53的负调节因子。本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，我们的初步数据表明， 表达突变体U2 AF 1的细胞转化为苏地霉素。我们假设RNA剪接机制的突变 通过使选择性剪接产物的表达失调而赋予剪接调节剂敏感性， 有助于MDS/AML发病机制。我们将在体外和体内使用定义明确的系统来检验这一假设。 vivo.在特定目标1中，基于以下研究，研究了具有或不具有剪接基因突变的原代人AML成髓细胞： 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）对体外生长、活力、凋亡和细胞周期动力学的影响， 免疫缺陷小鼠异种移植后体内嵌合、凋亡和三系分化 小鼠这将决定剪接基因突变是否作为对这些药物敏感的生物标志物。 在具体目标2中，我们将发现剪接调节剂诱导的转录组改变， 开发敏感的定量分析方法，以评估这些药剂的靶向效应。这 该项目通过识别预测以下疾病的遗传生物标志物来解决SPORE翻译终点： 对剪接调节剂的反应，并通过开发将用于药效学研究的测定， 剪接调节剂临床试验中的监测： 相关性（参见说明）： 目前的化疗治疗治愈了少数骨髓增生异常综合征（MDS）患者， 急性髓性白血病（AML）。我们和其他人最近发现，RNA剪接，一个关键的 代谢途径，在这些患者中的许多人中被改变，并且调节该途径的化合物是 available.该项目将测试这类新型化合物在人体细胞中的活性， 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.