Targeting FOXM1 in chemo-resistant monocytic AML

在过去的30年里，急性髓细胞白血病（AML）的预后一直很差。20-40%的 诱导化疗未能达到缓解的患者，50-70%达到缓解的患者， 3年内完全缓解复发。在剖析预后亚组方面的一个重大突破是 随着在40%-60%的细胞遗传学正常（CN）-AML中发现核磷蛋白（NPM 1）突变， 例在随后的分析中，已经表明具有野生型FMS样受体酪氨酸的AML患者， 携带突变的NPM 1（NPM 1 mut）的FLT 3激酶（FLT 3）显示出改善的总体存活和无复发存活。 我们提出，突变的NPM 1（NPM 1 mut）通过在AML-M5中螯合FOXM 1而在AML-M5中赋予这种优势。 FOXM 1失活的细胞质。在初步实验中，我们发现FOXM 1抑制剂： STL 427944（C25 H23 N7 O 4）和盐酸苯扎米尔（C13 H15 Cl 2N 7 O）抑制AML-M5中的核FOXM 1 细胞系我们提出以下具体目标：具体目标1。研究STL 427944的特异性 和盐酸苯扎米尔用于FOXM 1抑制及其在AML-M5细胞系中的抗癌活性。 首先，我们将使用单核细胞AML-M5细胞系THP 1、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FOXM 1。接下来，我们将用STL 427944处理AML-M5细胞， 盐酸苯扎明，并将使用RNA-seq比较亲代和治疗组的FOXM 1调控网络 细胞我们还将进行ChIP-seq实验，并通过比较来识别FOXM 1的直接靶点。 RNA-seq和ChIP-seq实验。因此，作为数据集成的结果，我们希望有一个优先级列表 属于FOXM 1网络的功能显著基因。具体目标2。以确定 在AML-M5小鼠模型中的新型FOXM 1抑制剂的药代动力学和功效。一是 测定STL 427944和盐酸苯扎明的毒性、药代动力学（PK）和生物利用度。到 我们将确定STL 427944/苯扎米尔在小鼠中的毒性以及药代动力学（PK）和生物利用度， 与毒理学研究实验室（TRL）合作，将在CD-1中检测STL 427944和苯扎明 小鼠将评估毒性和死亡的临床体征，持续2天，每天两次。为了确定 我们将用STL 427944/苯扎米尔作为小鼠中AML-M5的抗癌药物来治疗AML-M5异种移植肿瘤， STL 427944/苯扎米尔与阿糖胞苷或维奈托克组合的MTD。我们将建立几个小组， 使用不同AML-M5细胞系进行鼠AML异种移植，并将用STL 427944/苯扎米尔处理它们 盐酸盐单独或与阿糖胞苷或维奈托克组合。将监测小鼠12 将与溶媒治疗的患者进行比较，包括治疗时间、疾病表型、预期寿命和FOXM 1水平 小鼠这些实验对于确定是否可以进一步开发STL 427944/苯扎明至关重要 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.