Targeting FOXM1 in chemo-resistant monocytic AML

在过去的30年里，急性髓系白血病(AML)的预后一直很差。20%-40% 患者未能通过诱导化疗获得缓解，50%-70%的患者实现了 3年内完全缓解复发。在分离出预测亚群方面取得了重大突破 在40%-60%的细胞遗传学正常(CN)-AML中发现核磷蛋白(NPM1)突变 案子。随后的分析表明，具有野生型FMS样受体酪氨酸的AML患者 携带突变的NPM1(NPM1mut)的蛋白激酶(Flt3)基因可提高患者的总生存率和无复发生存率。 我们认为，突变的NPM1(NPM1mut)通过隔离FOXM1在AML-M5中赋予这一优势 FOXM1不活跃的细胞质。在初步实验中，我们表明FOXM1抑制剂： STL427944(C25H23N7O4)和苯扎米(C13H15Cl2N7O)对AML-M5细胞核FOXM1的抑制作用 细胞系。我们提出了以下具体目标：具体目标1.研究STL427944的特异性 和苯扎米对AML-M5细胞株FOXM1的抑制作用及其抗癌活性。 首先，我们将使用单核细胞AML-M5细胞株THP1，AML-193和U937来证实我们的初步数据 这些药物抑制AML-M5细胞系中的FOXM1。接下来，我们将用STL427944和STL427944处理AML-M5细胞 并将FOXM1调控网络在亲代和治疗中使用RNA-seq进行比较 细胞。我们还将进行芯片序列实验，并通过比较确定FOXM1的直接目标 RNA-SEQ和芯片-SEQ实验。因此，作为数据集成的结果，我们希望有一个按优先顺序排列的列表 属于FOXM1网络的重要功能基因。具体目标2.确定 新型FOXM1抑制剂在AML-M5小鼠模型中的药代动力学和疗效。首先，我们将 测定STL427944和盐酸苯扎米的毒性、药代动力学(PK)和生物利用度。至 测定STL427944/苯扎米在小鼠体内的毒性及药代动力学和生物利用度 与毒理学研究实验室(TRL)合作，STL427944和苯扎米将在CD-1中进行测试 老鼠。毒性和死亡率的临床体征将进行为期2天的评估，每天两次。确立…的效力 STL427944/苯扎米作为小鼠AML-M5的抗癌药物我们将对AML-M5移植瘤用 STL427944/苯扎米与阿糖胞苷或万乃馨联合应用的MTD。我们将建立几个小组 使用不同AML-M5细胞系的小鼠AML移植瘤，并将用STL427944/苯扎米处理它们 盐酸盐单独或与阿糖胞苷或文奈德合用。这些老鼠将被监测12年 周数和疾病表型，以及预期寿命和FOXM1水平将与车辆治疗进行比较 老鼠。这些实验将是确定STL427944/苯扎米能否进一步开发的关键 作为AML-M5患者的抗癌药物。

The outcomes for acute myeloid leukemia (AML) have remained poor for the past 30 years. 20- 40% of patients fail to achieve remission with induction chemotherapy, and 50-70% of patients who achieve a complete remission relapse within 3 years. A major breakthrough in dissecting out prognostic subgroups came with the discovery of the nucleophosmin (NPM1) mutation in 40%-60% of cytogenetically normal (CN)-AML cases. In subsequent analyses it has been shown that AML patients with wild-type FMS-like receptor tyrosine kinase (FLT3), bearing mutated NPM1 (NPM1mut) showed improved overall survival and relapse-free survival. We proposed that mutated NPM1 (NPM1mut) confers this advantage in AML-M5 via sequestration of FOXM1 in the cytoplasm where FOXM1 is inactive. In preliminary experiments we showed that FOXM1 inhibitors: STL427944 (C25H23N7O4) and benzamil hydrochloride (C13H15Cl2N7O) suppress nuclear FOXM1 in AML-M5 cell lines. We propose the following specific aims: Specific Aim 1. To investigate specificity of STL427944 and benzamil hydrochloride for FOXM1 suppression and their anticancer activity in AML-M5 cell lines. First, we will use monocytic AML-M5 cell lines THP1, AML-193 and U937 to confirm our preliminary data that these drugs inhibit FOXM1 in AML-M5 cell lines. Next, we will treat AML-M5 cells with STL427944 and benzamil hydrochloride and will compare FOXM1 regulatory network using RNA-seq in parental and treated cells. We will also perform ChIP-seq experiments and we will identify direct targets of FOXM1 by comparing RNA-seq and ChIP-seq experiments. Thus, as the result of data integration, we expect to have a prioritized list of functionally significant genes that belong to FOXM1 network. Specific Aim 2. To determine pharmacokinetics and efficacy of novel FOXM1 inhibitors in AML-M5 mouse models. First, we will determine toxicity, pharmacokinetics (PK) and bioavailability of STL427944 and benzamil hydrochloride. To determine toxicity of STL427944/benzamil in mice and also pharmacokinetics (PK) and bioavailability we will collaborate with the Toxicology Research Laboratory (TRL), STL427944 and benzamil will be tested in CD-1 mice. Clinical signs of toxicity and mortality will be assessed for 2 days, twice a day. To establish efficacy of STL427944/benzamil as anticancer drugs for AML-M5 in mice we will treat AML-M5 xenograft tumors with MTD of STL427944/benzamil in combination with cytarabine or venetoclax.. We will establish several groups of murine AML xenografts using different AML-M5 cell lines and will treat them with that STL427944/benzamil hydrochloride individually or in combination with cytarabine or venetoclax. The mice will be monitored for 12 weeks and disease phenotype, and life expectancy and FOXM1 levels will be compared with vehicle treated mice. These experiments will be crucial to determine whether STL427944/benzamil could be further developed as the anticancer drugs for AML-M5 patients.