Pathologic Signaling Pathways in AML Cells

AML 细胞中的病理信号通路

• 批准号: 10553601
• 负责人: MARTIN CARROLL
• 金额: --
• 依托单位: PHILADELPHIA VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10553601
• 关键词: Acute Myelocytic Leukemia; Affect; American; Binding; Biochemical Pathway; Biological Assay; Cell Count; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cell Survival; Clinical; Clinical Trials; Complex; Cytarabine; DNA; Data; Disease; Drug Combinations; Engraftment; Equilibrium; Etoposide; FLT3 gene; FLT3 inhibitor; FOXO3A gene; FRAP1 gene; Generations; Growth Factor; Hematopoietic Neoplasms; Human; IL3 Gene; In Vitro; Individual; Inositol; Laboratories; Lead; Leukemic Cell; Life; Metabolic; Methodology; Methods; Modeling; Mus; Mutate; PIK3CG gene; Pathogenesis; Pathologic; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Pre-Clinical Model; Process; Proteins; Proto-Oncogene Proteins c-akt; Regulation; Relapse; Reporting; Research Design; Research Proposals; Sampling; Series; Signal Pathway; Signal Transduction; Sirolimus; Therapeutic; Transcriptional Regulation; Translations; United States; United States National Institutes of Health; Veterans; Work; Xenograft Model; acute myeloid leukemia cell; c-myc Genes; cancer cell; cell growth; cell transformation; chemotherapy; chromatin immunoprecipitation; combinatorial; cytotoxic; improved; in vivo; inhibitor; insulin signaling; leukemic stem cell; molecular subtypes; novel strategies; novel therapeutics; promoter; small molecule inhibitor; success

Abstract: Acute myeloid leukemia (AML) is a serious blood cancer that affects United States Veterans and other individuals. Multiple studies have demonstrated that active signaling pathways are a necessary step to leukemic cell transformation. However, the mechanism(s) of activated signaling in FLT3 wild type AML are poorly described. We hypothesize that constitutive activation of the PI3 kinase pathway leads to activation of a complex called mammalian target of rapamycin 2 (mTORC2) which regulates diverse pathways to balance cell growth and survival in AML cells. In this proposal, we will define these signaling pathways and determine if inhibition of mTORC2 alone or in combination with other drugs will enhance survival in pre-clinical models of AML leading to new therapies for this challenging disease. 1. Objective(s): The overall objective of the research proposal is to define the mechanisms of activated cell signaling in AML cells, determine the biochemical pathways regulated by mTORC2 in AML cells and determine if inhibition of mTORC2 in pre-clinical models can increase survival of individuals with AML. 2. Research Design: We propose three Specific Aims. Specific Aim 1) Confirm that 4EBP1 is a target of MTORC2 and not MTORC1 in human AML samples and define whether MTORC2 regulates cell survival or cell growth. Specific Aim 2) Determine if mTORC2 Regulates FOXO 3 phosphorylation in AML cells to Regulate Cell Survival. Specific Aim 3) Determine if combinatorial MTORC1/MTORC2 inhibition in combination with chemotherapy or Bcl2 inhibition suppresses AML cell growth in pre- clinical models. 3. Methodology: Specific Aim 1 will primary use human AML cells and primary human AML cells in culture. Specific Aim 2 will use similar methods but also assess FOXO binding to DNA using chromatin immunoprecipitation. Specific Aim 3 will focus on studies in a xenotransplantation model of AML developed by our laboratory. 4. Findings: We anticipate confirming that mTORC2 is actually a master regulator of signaling in AML cells. In particular, we anticipate that mTORC2 regulates 4EBP1 to regulate protein translation. We also anticipate that mTORC2 regulates FOXO proteins to regulate c-Myc expression. Finally, we anticipate that mTORC2 inhibition will work alone or in combination with other therapies to inhibit AML cell survival in vivo. 5. Clinical Relationships: These studies will use primary material from patients with AML and will be used to develop a new approach to therapy of the disease.

【摘要】：急性髓系白血病(AML)是一种严重影响人类健康的血液癌症。 各州退伍军人和其他个人。多项研究表明，主动 信号通路是白血病细胞转化的必要步骤。然而， FLT3 野生型 AML 中激活信号传导的机制描述很少。我们 假设 PI3 激酶途径的组成型激活导致 称为哺乳动物雷帕霉素靶标 2 (mTORC2) 的复合体，可调节多种 平衡 AML 细胞生长和存活的途径。在本提案中，我们将 定义这些信号通路并确定是否单独抑制 mTORC2 或联合抑制 mTORC2 与其他药物联合使用将提高 AML 临床前模型的生存率 导致针对这种具有挑战性的疾病的新疗法。 1. 目标：研究计划的总体目标是定义 AML 细胞中激活的细胞信号传导机制，确定生化 AML 细胞中 mTORC2 调节的通路并确定 mTORC2 的抑制是否有效 在临床前模型中可以提高 AML 患者的生存率。 2. 研究设计：我们提出了三个具体目标。具体目标 1) 确认 4EBP1 是人类 AML 样本中 MTORC2 而非 MTORC1 的靶标，并定义 MTORC2 是否调节细胞存活或细胞生长。具体目标 2) 确定是否 mTORC2 调节 AML 细胞中的 FOXO 3 磷酸化以调节细胞存活。 具体目标 3) 确定 MTORC1/MTORC2 组合抑制是否 与化疗或 Bcl2 抑制剂联合使用可抑制 AML 细胞生长 临床模型。 3. 方法：具体目标 1 将主要使用人类 AML 细胞和主要 培养中的人类 AML 细胞。具体目标 2 将使用类似的方法，但也会评估 使用染色质免疫沉淀法将 FOXO 与 DNA 结合。具体目标3将重点关注 对我们实验室开发的 AML 异种移植模型进行研究。 4. 研究结果：我们预计确认 mTORC2 实际上是 AML 细胞中的信号传导。特别是，我们预计 mTORC2 调节 4EBP1 调节蛋白质翻译。我们还预计 mTORC2 调节 FOXO 蛋白 调节 c-Myc 表达。最后，我们预计 mTORC2 抑制将会发挥作用 单独或与其他疗法联合使用可抑制体内 AML 细胞存活。 5. 临床关系：这些研究将使用来自患有以下疾病的患者的主要材料： AML 将用于开发治疗该疾病的新方法。