Cytoplasmic FOXM1 contributes to the higher complete remission and longer overall survival of AML patients after chemotherapy

细胞质 FOXM1 有助于 AML 患者化疗后更高的完全缓解率和更长的总生存期

• 批准号: 8876933
• 负责人: ANDREI L GARTEL
• 金额: $ 20.85万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8876933
• 关键词: Accounting; Acute Myelocytic Leukemia; Affect; Allogenic; Apoptosis; Archives; Biological Markers; Blast Cell; Bone Marrow; Bone Marrow Transplantation; Categories; Cell Communication; Cell Line; Cell Nucleus; Cell Proliferation; Cells; Characteristics; Chromosome abnormality; Classification; Clinical; Cytarabine; Cytogenetics; Cytoplasm; Data; Development; Diagnosis; Diagnostic; Disease; Disease remission; Doxorubicin; Gene Expression Profiling; Genes; Goals; Grant; HL-60 Cells; HL60; Hematologic Neoplasms; Human; Immunofluorescence Immunologic; In Vitro; Laboratories; Link; Malignant Neoplasms; Molecular; Molecular Profiling; Mus; Mutate; Mutation; NPM1 gene; Neoplasms; Nuclear; Nuclear Export; Nude Mice; Oncogenic; One-Step dentin bonding system; Outcome; Pathologic; Patients; Pharmaceutical Preparations; Phenotype; Prognostic Factor; Prognostic Marker; Property; Proteins; RNA Interference; Recurrence; Relapse; Resistance; Risk; Role; Sampling; Stratification; Subgroup; Testing; Time; Tissues; Transplantation; U937 Cells; Vertebral column; Xenograft procedure; biobank; cancer cell; chemotherapy; clinically significant; cohort; imaging modality; imaging system; improved; in vivo; leukemia; liquid crystal polymer; mouse model; mutant; novel; novel strategies; novel therapeutics; nucleophosmin; outcome forecast; overexpression; prognostic; public health relevance; research study; response; transcription factor; treatment strategy; tumor; tumor xenograft

 DESCRIPTION (provided by applicant): The outcomes for acute myeloid leukemia (AML) have remained abysmal 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. While cytogenetics are the backbone of risk-stratification, nearly 50% of AML cases are cytogenetically normal (CN-AML) and, biologically and clinically, the most heterogenous group. The nucleo-cytoplasmic shuttle protein nucleophosmin (NPM1) is mutated in 50% of CN-AML cases resulting in the nuclear export of this protein. This has been shown in large clinical cohorts to confer a favorable prognosis and obviates the need for a bone marrow transplant in these patients. This subset of AML has been recognized in a unique diagnostic category in the WHO 2008 classification of hematologic malignancies. The mechanism by which mutated NPM1 (NPM1mut) confers this advantage in CN-AML is unclear. It has previously been hypothesized that NPM1mut dislocates other protein partners into the cytoplasm and consequently affecting their function. We have previously demonstrated that FOXM1, an oncogenic transcription factor, co-localizes with NPM1 in cancer cells. In this proposal, using a novel multispectral imaging modality we will study the cellular interaction of FOXM1 and NPM1 in AML primary blast cells. We will also investigate the functional significance of FOXM1 localization in this disease using AML cell lines. Importantly, we will assess the clinical correlation of mislocalized cytoplasmic FOXM1 with outcomes in AML with the goal of examining its role as a favorable prognostic marker in CN-AML. We intend to show that FOXM1 is the oncogenic driver dictating prognosis in AML. Mutations in NPM1 resulting in its nuclear export mislocalizes FOXM1 to the cytoplasm where it is inactive as a transcription factor. This may account for the improved outcome in this sizeable subset of AML patients. In several AML cell lines we will test how FOXM1 knockdown or FOXM1 overexpression will affect sensitivity to chemotherapeutic drugs in vitro and vivo. We will determine whether FOXM1 knockdown in AML cell lines with nuclear localization of FOXM1 makes them more sensitive to chemotherapy. Nude mice will be injected with AML cell lines with stable FOXM1 knockdown or OCI-AML3 cell line with stable FOXM1 knockdown or with FOXM1 overexpression, and with control cell lines to establish xenograft tumors. We expect that FOXM1 knockdown in U937 and HL60 AML cell lines will increase sensitivity to treatment in xenograft tumors induced by these AML cell lines. I contrast, suppression of FOXM1, localized in the cytoplasm, in OCI-AML3 cell line would not affect sensitivity to chemotherapy in xenograft tumors induced by this cell line. The experiments described in our proposal will reveal if cytoplasmic FOXM1 is a novel favorable prognostic factor in AML and whether nuclear expression of FOXM1 determines the resistance of xenograft tumors induced by AML cell lines to chemotherapy.

 描述（由申请人提供）：在过去的30年里，急性髓性白血病（AML）的结局一直很糟糕。20-40%的患者在诱导化疗后未能达到缓解，50-70%达到完全缓解的患者在3年内复发。虽然细胞遗传学是风险分层的支柱，但近50%的AML病例细胞遗传学正常（CN-AML），并且在生物学和临床上是最异质的组。核质穿梭蛋白核磷蛋白（NPM 1）在50%的CN-AML病例中发生突变，导致该蛋白质的核输出。这已在大型临床队列中显示出有利的预后，并避免了这些患者对骨髓移植的需要。AML的这一亚组在WHO 2008血液系统恶性肿瘤分类中被确认为独特的诊断类别。突变的NPM 1（NPM 1 mut）在CN-AML中赋予这种优势的机制尚不清楚。以前曾假设NPM 1 mut将其他蛋白质伴侣移位到细胞质中，从而影响它们的功能。我们以前已经证明，FOXM 1，一种致癌转录因子，与NPM 1在癌细胞中共定位。在这个建议中，使用一种新的多光谱成像模式，我们将研究AML原代母细胞中FOXM 1和NPM 1的细胞相互作用。我们还将使用AML细胞系研究FOXM 1定位在这种疾病中的功能意义。重要的是，我们将评估错误定位的细胞质FOXM 1与AML结局的临床相关性，目的是检查其作为CN-AML有利预后标志物的作用。我们打算证明FOXM 1是决定AML预后的致癌驱动因子。NPM 1突变导致其核输出，将FOXM 1错误定位到细胞质中，在细胞质中它作为转录因子是无活性的。这可能是这一相当大的AML患者亚组结局改善的原因。在几种AML细胞系中，我们将测试FOXM 1敲低或FOXM 1过表达如何影响体外和体内化疗药物的敏感性。我们将确定在具有FOXM 1核定位的AML细胞系中FOXM 1敲低是否使它们对化疗更敏感。将用具有稳定的FOXM 1敲低的AML细胞系或具有稳定的FOXM 1敲低或具有FOXM 1过表达的OCI-AML 3细胞系以及对照细胞系注射裸鼠以建立异种移植肿瘤。我们预计，在U937和HL 60 AML细胞系中的FOXM 1敲低将增加由这些AML细胞系诱导的异种移植肿瘤对治疗的敏感性。相反，在OCI-AML 3细胞系中抑制位于细胞质中的FOXM 1不会影响由该细胞系诱导的异种移植肿瘤对化疗的敏感性。在我们的提案中描述的实验将揭示胞质FOXM 1是否是AML中的一种新的有利预后因素，以及FOXM 1的核表达是否决定了AML细胞系诱导的异种移植肿瘤对化疗的抗性。