Inhibition of Pim kinases in acute myeloid leukemia

急性髓性白血病中 Pim 激酶的抑制

• 批准号: 8733336
• 负责人: MARIA R BAER
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-10-01 至 2018-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8733336
• 关键词: ABCB1 gene; ABCC1 gene; ABCG2 gene; Accounting; Acute Myelocytic Leukemia; Acute leukemia; Adult; Age; Anthracyclines; Antimetabolites; Apoptosis; Apoptotic; Applications Grants; Binding; Cell Cycle; Cell Line; Cells; Clinical; Clinical Trials; Cytarabine; DNA Damage; DNA Repair; Daunorubicin; Development; Diagnosis; Disease; Disease Progression; Disease Resistance; Down-Regulation; Drug effect disorder; Drug resistance; Drug usage; Environmental Exposure; Exposure to; Feedback; Female; Generations; Genes; Genomic Instability; Goals; Health; Hematopoietic; Hematopoietic Stem Cell Transplantation; In Vitro; Incidence; Induction of Apoptosis; Kinetics; Lead; Ligands; MCL1 gene; Malignant Neoplasms; Mediating; Medical; Military Personnel; Molecular Abnormality; Multi-Drug Resistance; Mutation; Nonhomologous DNA End Joining; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Phosphorylation; Play; Population; Protein-Serine-Threonine Kinases; Proteins; Radiation therapy; Reactive Oxygen Species; Receptor Protein-Tyrosine Kinases; Receptor Tyrosine Kinase Gene; Regimen; Relapse; Reporting; Resistance; Sampling; Schedule; Services; Signal Transduction; Stem cells; System; Testing; Therapeutic Index; Treatment Protocols; Treatment outcome; Up-Regulation; Vascular Endothelial Growth Factor Receptor-1; Veterans; Work; chemotherapy; design; disorder risk; immunodeficient mouse model; improved; in vitro testing; in vivo; in vivo Model; inhibitor/antagonist; insight; kinase inhibitor; leukemia treatment; leukemic stem cell; male; overexpression; pre-clinical; pro-apoptotic protein; proto-oncogene protein pim; proto-oncogene protein pim-1; repaired; resistance mechanism; response; uptake

DESCRIPTION (provided by applicant): Acute myeloid leukemia (AML) is an important health concern for veterans, due to its high incidence and frequent poor response to treatment. Notably, internal tandem duplication (ITD) mutations of the fms-like tyrosine kinase 3 (FLT3) receptor tyrosine kinase genes are present AML cells in a third of AML patients and are associated with rapid relapse following chemotherapy. FLT3 inhibitors have activity, but responses are limited and transient. Pim-1 kinase is a serine threonine kinase that is expressed in AML cells and is overexpressed downstream of FLT3-ITD in AML cells with this molecular abnormality. Pim-1 kinase regulates diverse proteins involved in proliferation, cell cycle, apoptosis and drug resistance, including, among others, c-MYC, the pro-apoptotic protein Bad and the anti-apoptotic protein Mcl-1, and is also implicated in DNA repair. We demonstrated that Pim-1 also phosphorylates FLT3 and promotes its aberrant signaling in a positive feedback loop in FLT3-ITD AML cells, and that Pym kinase inhibitors sensitize FLT3-ITD AML cells to apoptosis induction by FLT3 inhibitors. We then found that Pim kinase inhibitors also sensitize FLT3-ITD AML cells to apoptosis induction by chemotherapy drugs used to treat AML. Pim-1 substrate proteins are also expressed in key pathways in AML stem cells, and Pim-1 phosphorylation of these proteins may protect AML stem cells from the effects of chemotherapy and of FLT3 inhibitors. Pim kinase inhibitors are in current preclinical and clinical development. 1. The overall hypotheses of this grant proposal is that Pim-1 plays a key role in resistance of FLT3-ITD AML to available treatments, and that inhibition of Pim kinase will improve treatment outcomes in FLT3-ITD AML by abrogating multiple mechanisms of drug resistance and of disease progression. The Specific Aims of the proposed work are: 1. To determine the mechanisms by which Pim kinase inhibition sensitizes FLT3-ITD AML cells to induction of apoptosis by chemotherapy drugs and by FLT3 inhibitors; 2. To optimize scheduling of administration of Pim kinase inhibitors, chemotherapy drugs and by FLT3 inhibitors in FLT3-ITD AML; and 3. To test the effects of in vivo administration of Pim kinase inhibitors with chemotherapy drugs and with FLT3 inhibitors on FLT3-ITD AML cells and AML stem cells and on normal hematopoietic cells. The effects of Pim-1 kinase inhibition will be studied in AML cell lines and patient samples using both in vitro culture systems and an in vivo model. In Aim 1, we will test the hypotheses that Pim kinase inhibition sensitizes FLT3-ITD cells to apoptosis by inhibiting FLT3-ITD signaling, altering expression and activation of pro- and anti-apoptotic proteins, promoting generation of reactive oxygen species and/or inhibiting repair of DNA damage. Mechanistic insights achieved in Aim 1 will guide design of scheduling of Pim kinase inhibition in relation to exposure to chemotherapy drugs and to FLT3 inhibitors. Approaches to scheduling will be tested in vitro, with apoptosis as the readout. In addition, effects of Pim kinase inhibition on cell cycle could result in kinetic resistance to drugs, and notably cytarabine, and will be tested. Finally, effects on normal hematopoietic cells will be studied to establish a therapeutic index. In Aim 3, strategies optimized in vitro in Aim 2 will be tested in vivo an immunodeficient mouse model. It is anticipated that the proposed work will lead to development and optimization of regimens incorporating Pim kinase inhibitors in the treatment of FLT3-ITD AML, with the goal of improved outcomes in this unfavorable AML subset...

描述（由申请人提供）： 急性髓系白血病（AML）因其发病率高且治疗反应经常不佳而成为退伍军人的一个重要健康问题。值得注意的是，fms 样酪氨酸激酶 3 (FLT3) 受体酪氨酸激酶基因的内部串联重复 (ITD) 突变存在于三分之一的 AML 患者的 AML 细胞中，并且与化疗后的快速复发相关。 FLT3 抑制剂具有活性，但反应有限且短暂。 Pim-1 激酶是一种丝氨酸苏氨酸激酶，在 AML 细胞中表达，并且在具有这种分子异常的 AML 细胞中 FLT3-ITD 下游过表达。 Pim-1 激酶调节涉及增殖、细胞周期、凋亡和耐药性的多种蛋白，其中包括 c-MYC、促凋亡蛋白 Bad 和抗凋亡蛋白 Mcl-1，并且还参与 DNA 修复。我们证明，Pim-1 还可磷酸化 FLT3，并在 FLT3-ITD AML 细胞的正反馈环中促进其异常信号传导，并且 Pym 激酶抑制剂使 FLT3-ITD AML 细胞对 FLT3 抑制剂诱导的细胞凋亡敏感。然后我们发现 Pim 激酶抑制剂还可使 FLT3-ITD AML 细胞对用于治疗 AML 的化疗药物诱导的细胞凋亡敏感。 Pim-1 底物蛋白也在 AML 干细胞的关键通路中表达，这些蛋白的 Pim-1 磷酸化可以保护 AML 干细胞免受化疗和 FLT3 抑制剂的影响。 Pim 激酶抑制剂目前正处于临床前和临床开发阶段。 1. 本拨款提案的总体假设是，Pim-1 在 FLT3-ITD AML 对现有治疗的耐药性中发挥着关键作用，并且抑制 Pim 激酶将通过消除多种耐药性和疾病进展机制来改善 FLT3-ITD AML 的治疗结果。拟议工作的具体目标是： 1. 确定 Pim 激酶抑制使 FLT3-ITD AML 细胞对化疗药物和 FLT3 抑制剂诱导凋亡敏感的机制； 2. 优化FLT3-ITD AML中Pim激酶抑制剂、化疗药物和FLT3抑制剂的给药安排； 3.测试体内给予Pim激酶抑制剂与化疗药物和FLT3抑制剂对FLT3-ITD AML细胞和AML干细胞以及对正常造血细胞的影响。 将使用体外培养系统和体内模型在 AML 细胞系和患者样本中研究 Pim-1 激酶抑制的效果。在目标 1 中，我们将测试以下假设：Pim 激酶抑制通过抑制 FLT3-ITD 信号传导、改变促凋亡蛋白和抗凋亡蛋白的表达和激活、促进活性氧的产生和/或抑制 DNA 损伤的修复来使 FLT3-ITD 细胞对凋亡敏感。目标 1 中实现的机制见解将指导与化疗药物和 FLT3 抑制剂暴露相关的 Pim 激酶抑制的安排设计。调度方法将在体外进行测试，以细胞凋亡作为读数。此外，Pim 激酶抑制对细胞周期的影响可能会导致对药物（特别是阿糖胞苷）的动力学耐药性，并将进行测试。最后，将研究对正常造血细胞的影响以确定治疗指数。在目标 3 中，目标 2 中体外优化的策略将在免疫缺陷小鼠模型中进行体内测试。预计拟议的工作将导致开发和优化结合 Pim 激酶抑制剂治疗 FLT3-ITD AML 的方案，目标是改善这一不利的 AML 子集的结果......