A rapid spontaneous murine model of CN-AML

摘要 急性髓系白血病(AML)基因复杂，但患者可分为有染色体的患者 易位和细胞遗传学正常的(CN-AML)。CN-AML占人类总数的近50% 急性髓系白血病患者中，成人CN-AML患者的总5年生存率约为30%。De基因的突变 Novo DNA甲基转移酶DNMT3A与FMS样酪氨酸激酶3的内部串联复制 (Flt3-ITD)和是CN-AML中最常见的两个事件。此外，最近的全基因组测序 在CN-AML患者样本中发现：1)Flt3和DNMT3A突变的高频共存，以及 2)CN-AML患者的风险分类相应改变，预后较差。因为 人类流行病学研究不能轻易控制基因工程带来疾病风险的因素 小鼠(GEM)品系为机械剖析疾病病理生物学提供了必要的易处理平台， 异质性和治疗反应。虽然Flt3-ITD和DNMT3A的可诱导缺失都不会导致 当我们将Flt3-ITD突变等位基因与DNMT3A的诱导缺失相结合时，小鼠的自发性白血病 我们发现了一种自发性、快速致命性、完全穿透性和可移植的AML。我们假设 在Flt3-ITD小鼠中可诱导的DNMT3A缺失产生了一种忠实的人CN-AML模型，可用于 推断构成治疗反应的基本生物学和分子因素。为此，我们建议， 基因组、细胞遗传学和单细胞分子分析(与原代人类CN-AML比较) 解开肿瘤的结构和潜在的细胞状态。我们期待着拟议的研究 提供经验证的Flt3-ITD/DNMT3a突变型CN-AML小鼠模型，并定义翻译实用程序。

ABSTRACT Acute myeloid leukemia (AML) is genetically complex, but patients can be divided into those with chromosomal translocations and those that are cytogenetically normal (CN-AML). CN-AML represents nearly 50% of human AML cases, and the overall 5-year survival for adults with CN-AML is approximately 30%. Mutations in the de novo DNA methyltransferase DNMT3A and internal tandem duplications of the FMS-like tyrosine kinase 3 (FLT3-ITD) and are two of the most frequent events in CN-AML. Moreover, recent whole-genome sequencing of CN-AML patient samples identified: 1) high-frequency co-occurrence of FLT3 and DNMT3A mutations, and 2) corresponding changes to the risk classification for these CN-AML patients to a poorer prognosis. Because human epidemiologic studies cannot easily control for factors that confer disease risk, genetically engineered mouse (GEM) strains provide essential tractable platforms to mechanistically dissect disease pathobiology, heterogeneity and therapeutic response. Although neither Flt3-ITD nor inducible deletion of Dnmt3a induces spontaneous leukemia in mice, when we combined Flt3-ITD mutant alleles with inducible deletion of Dnmt3a we find a spontaneous, rapidly lethal, completely-penetrant, and transplantable AML. We hypothesize that inducible deletion of Dnmt3a in Flt3-ITD mice produces a faithful model of human CN-AML that can be used to infer essential biological and molecular factors that constitute therapeutic response. To this end, we propose, genomic, cytogenetic, and single-cell molecular analyses (with comparison to primary human CN-AML) to deconvolute both the tumor architecture and the underlying cellular states. We expect the proposed research to deliver a validated murine model of FLT3-ITD/DNMT3a-mutant CN-AML with defined translational utility.