Targeting FOXM1 in chemo-resistant monocytic AML

靶向 FOXM1 治疗耐药单核细胞 AML

• 批准号: 10187213
• 负责人: ANDREI L GARTEL
• 金额: $ 26.38万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10187213
• 关键词: 11q23; Acute Myelocytic Leukemia; Acute leukemia; Acute monocytic leukemia; Acute respiratory failure; Adult; Antineoplastic Agents; Apoptosis; Biological Availability; Blood Coagulation Disorders; CASP3 gene; Cell Line; Cells; ChIP-seq; Characteristics; Chemoresistance; Cleaved cell; Clinical; Cytarabine; Cytogenetics; Cytoplasm; Data; Disease remission; Dose; Drug Kinetics; FOXM1 gene; Family; Genes; Gingival Hyperplasia; Immunoblotting; Individual; KRAS2 gene; Laboratory Research; Leukemic Cell; Libraries; Life Expectancy; Link; Mammalian Cell; Maximum Tolerated Dose; Molecular Chaperones; Monitor; Monocytic leukemia; Mus; Mutate; Mutation; NPM1 gene; Nature; Neuraxis; Nuclear; Outcome; Patients; Pharmaceutical Preparations; Preparation; Prognosis; Promonocyte; Protein Analysis; Proteins; Protocols documentation; RNA; Receptor Protein-Tyrosine Kinases; Relapse; Sampling; Specificity; Subgroup; Tail; Testing; Tissues; Toxic effect; Toxicology; Ursidae Family; Veins; Visceromegaly; Whole-Body Irradiation; Xenograft procedure; acute myeloid leukemia cell; anticancer activity; base; benzamil; beta Actin; chemotherapy; data integration; differential expression; disease phenotype; experimental study; improved; improved outcome; inhibitor/antagonist; leukemia; lymphadenopathy; monoblast; monocyte; mortality; mouse model; mutant; novel; nucleophosmin; nucleoplasmin; prognostic; small molecule; synergism; transcriptome sequencing; tumor xenograft

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.

在过去的30年中，急性髓样白血病（AML）的结果一直保持较差。 20-40％ 患者无法通过诱导化疗来缓解，而50-70％的患者达到了 在3年内完全缓解复发。剖析预后亚组的主要突破来了 在40％-60％的细胞遗传学正常（CN）-AML中发现了核素（NPM1）突变 案例。在随后的分析中，已经显示出具有野生型FMS样受体酪氨酸的AML患者 激酶（FLT3），轴承突变的NPM1（NPM1MUT）表现出改善的总生存率和无复发生存率。 我们提出，突变的NPM1（NPM1MUT）通过封存FOXM1在AML-M5中赋予了这一优势。 FOXM1不活跃的细胞质。在初步实验中，我们表明FOXM1抑制剂： STL427944（C25H23N7O4）和盐酸苯甲酰基（C13H15CL2N7O）抑制AML-M5中的核FOXM1 细胞系。我们提出以下特定目的：特定目的1。研究STL427944的特异性 和盐酸苯甲胺盐抑制及其在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THP1，AML-193和U937来确认我们的初步数据 这些药物抑制AML-M5细胞系中的FOXM1。接下来，我们将使用STL427944处理AML-M5细胞 盐酸苯甲胺盐，并将在父母中使用RNA-Seq比较FOXM1调节网络 细胞。我们还将执行CHIP-SEQ实验，我们将通过比较FOXM1的直接目标 RNA-Seq和ChIP-Seq实验。因此，由于数据集成，我们希望有优先的列表 属于FOXM1网络的功能重要基因。特定目标2。确定 新型FOXM1抑制剂在AML-M5小鼠模型中的药代动力学和功效。首先，我们会的 确定毒性，药代动力学（PK）和STL427944和盐酸苯甲胺的毒性。到 确定STL427944/Benzamil在小鼠以及药代动力学（PK）和生物利用度中的毒性，我们将 与毒理学研究实验室（TRL），STL427944和Benzamil合作将在CD-1中进行测试 老鼠。每天两次，将评估毒性和死亡率的临床迹象2天。建立功效 STL427944/Benzamil作为小鼠AML-M5的抗癌药 STL427944/Benzamil的MTD与Cytarabine或Venetoclax结合使用。我们将建立几组的 使用不同的AML-M5细胞系使用鼠AML异种移植物，并将用STL427944/Benzamil对其进行处理 盐酸盐单独或与黄细胞捕滨或venetoclax结合使用。小鼠将被监测12 几周和疾病表型，预期寿命和FOXM1水平将与经过处理的车辆进行比较 老鼠。这些实验对于确定STL427944/Benzamil是否可以进一步发展至关重要 作为AML-M5患者的抗癌药物。