Validation and Development of WEE1 as a therapeutic target in AML

WEE1 作为 AML 治疗靶点的验证和开发

• 批准号: 8688970
• 负责人: CHRISTOPHER C PORTER
• 金额: $ 30.56万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8688970
• 关键词: Acute Myelocytic Leukemia; Animal Model; Anthracyclines; Antimetabolites; Antineoplastic Agents; Biological Assay; Biological Markers; Biopsy Specimen; Blast Cell; Bone marrow biopsy; CDC2 Protein Kinase; Cell Cycle Checkpoint; Cell Death; Cell Line; Cell Survival; Cells; Clinic; Clinical Trials; Combination Drug Therapy; Combined Modality Therapy; Cytarabine; Cytotoxic Chemotherapy; DNA Damage; DNA Repair; DNA biosynthesis; Data; Development; Disease remission; Dose; Drug Targeting; Ensure; Event; FLT3 gene; Failure; Genetic; Goals; Heterogeneity; Immunohistochemistry; Knowledge; Lead; Mediator of activation protein; Methodology; Methods; Mitosis; Modeling; Molecular; Molecular Abnormality; Mutation; Myeloid Progenitor Cells; Oncogenic; Outcome; Patients; Pharmaceutical Preparations; Pharmacodynamics; Phase; Proteins; Public Health; Recovery; Relapse; Remission Induction; Research; Resistance; Role; S Phase; Stress; Surveys; Survival Rate; Testing; Therapeutic; Time; Translating; Translations; Treatment Protocols; Validation; Work; Xenograft procedure; cancer diagnosis; cancer therapy; cell killing; chemotherapy; combinatorial; cytotoxic; design; ds-DNA; genome-wide; improved; in vivo; in vivo Model; inhibitor/antagonist; innovation; leukemia; leukemogenesis; mouse model; new therapeutic target; novel; novel therapeutics; pre-clinical; public health relevance; response; small hairpin RNA; therapeutic target; treatment strategy

DESCRIPTION (provided by applicant): Survival rates for patients with acute myeloid leukemia (AML) remain low, despite the fact that knowledge of the molecular events that cause AML has evolved rapidly in the last decade. This is in part due to a distinct lack of understanding of the response to chemotherapy at the cellular and molecular levels. Previous work has been directed toward the long term goal of developing novel, rational combination-therapies to improve the outcomes of patients with AML. The objective of this specific project is to provide pre-clinical validation of one such therapeutic strategy, combining antimetabolite chemotherapeutics with inhibition of Wee1, a cell cycle checkpoint protein. The central hypothesis of this project is that Wee1 is a critical mediator of leukemia cell survival, acting inS phase to ensure accurate DNA replication, and that combining inhibition of Wee1 with antimetabolites will be more effective in eliminating leukemia cells than antimetabolites alone. This hypothesis emanates from our preliminary data from a genome-wide shRNA screen, in which we identified Wee1 as a critical mediator of AML cell death after cytarabine treatment. We have validated these findings in a broad panel of AML cell lines and found that the effect of inhibiting Wee1 in combination with cytarabine involves abrogation of the S phase checkpoint induced by cytarabine. Notably, preliminary data show that cells with Class I leukemogenic mutations are quite sensitive to Wee1 inhibition alone and suggest that Wee1 may contribute to leukemogenesis. The hypothesis will be tested with three specific aims: 1) Determine whether Wee1 contributes to leukemogenesis, 2) Confirm the efficacy and tolerability of cytarabine and Wee1 inhibition in vivo and 3) Identify the mechanism of enhanced AML cell killing with cytarabine and Wee1 inhibition. To achieve Aim 1, we will examine the consequences of Class I and II mutation expression and Wee1 inhibition as they relate to oncogenic stresses in myeloid progenitor cells and perform immunohistochemistry for Wee1 expression in AML bone marrow biopsy specimens. To achieve Aim 2, mouse models of AML, including direct patient xenografts will be used to test the pharmacodynamics, tolerability and efficacy of combining cytarabine with Wee1 inhibition. To achieve Aim 3, pharmacologic and genetic inhibition will be used to test the functional significance of proteins upstream, downstream and in parallel with Wee1 in cell cycle checkpoint cascades to identify the key molecules involved; complementary cellular and molecular assays will be used to further define the mechanism of combinatorial cell killing. The project is innovative in 1) the study of Wee1 contributing to leukemogenesis, 2) the study of non-oncogene vulnerabilities of FLT3-ITD+ cells, 3) the development of a novel therapeutic target in AML, 4) translational modeling of leukemia with direct patient xenografts, and 5) high-throughput methodology to survey the role of DNA damage repair proteins in specific contexts. The proposed research is significant because it is expected to advance the understanding of a novel therapeutic target in AML that can be rapidly translated into clinical trials combining standard and targeted therapy.

描述（由申请人提供）：尽管了解导致AML的分子事件的知识在过去十年中迅速发展，但急性髓样白血病（AML）患者的存活率仍然很低。这部分是由于对细胞和分子水平上化学疗法反应的明显了解。先前的工作已针对开发新颖的理性组合 - 治疗措施以改善AML患者的预后的长期目标。该特定项目的目的是对一种这种治疗策略进行临床前验证，将抗代谢物化学治疗药与抑制WEE1（细胞周期检查点蛋白质）相结合。该项目的中心假设是WEE1是白血病细胞存活的关键介体，作用INS相，以确保准确的DNA复制，并且将WEE1与抗代谢物的抑制相结合将比单独消除白血病细胞更有效。该假设从我们的初步数据中源自全基因组shRNA筛查，在该数据中，我们将WEE1鉴定为细胞治疗后AML细胞死亡的关键介体。我们已经在宽阔的AML细胞系中验证了这些发现，发现抑制WEE1与细胞链滨组合的效果涉及废除由Cytarabine诱导的S相检查点。值得注意的是，初步数据表明，具有I类白血病性突变的细胞仅对WEE1抑制非常敏感，并表明WEE1可能有助于白血病。该假设将以三个特定的目的进行检验：1）确定WEE1是否有助于白血病发生，2）确认细胞中胞滨和WEE1抑制的疗效和耐受性，3）确定通过cytarabine和WEE1抑制作用增强AML细胞杀死的机制。为了实现目标1，我们将检查I类突变表达和WEE1抑制的后果，因为它们与髓样祖细胞中的致癌应激相关，并在AML骨髓活​​检标本中进行WEE1表达进行免疫组织化学。为了达到目标2，AML的小鼠模型（包括直接患者异种移植物）将用于测试将细胞动力学，耐受性和将Cytarabine与WEE1抑制相结合的功效。为了达到目标3，将使用药物和遗传抑制作用来测试上游，下游并与W​​EE1在细胞周期检查点级联反应中的功能意义，以识别所涉及的关键分子；互补的细胞和分子测定将用于进一步定义组合细胞杀死的机制。 The project is innovative in 1) the study of Wee1 contributing to leukemogenesis, 2) the study of non-oncogene vulnerabilities of FLT3-ITD+ cells, 3) the development of a novel therapeutic target in AML, 4) translational modeling of leukemia with direct patient xenografts, and 5) high-throughput methodology to survey the role of DNA damage repair proteins in specific contexts.拟议的研究很重要，因为预计它将提高对AML中新型治疗靶点的理解，该靶标可以迅速转化为结合标准和靶向治疗的临床试验。