Molecular mechanisms of drug resistance and disease progression in acute myeloid leukemia

急性髓系白血病耐药和疾病进展的分子机制

• 批准号: 9922661
• 负责人: Elizabeth Ann Eklund
• 金额: --
• 依托单位: JESSE BROWN VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9922661
• 关键词: 11q23; Acute Myelocytic Leukemia; Alkylating Agents; Anabolism; Apoptotic; Blocking Antibodies; Bone Marrow; Bone Marrow Cells; CD34 gene; CREBBP gene; Cells; Chimeric Proteins; Chromosome 7; Chromosome abnormality; Clinical; Complex; Cytogenetics; Cytokine Receptors; Cytokine Signaling; Cytotoxic Chemotherapy; Disease; Disease Progression; Disease remission; Drug resistance; Dysmyelopoietic Syndromes; Emergency Situation; Equilibrium; Event; Expression Profiling; FLT3 gene; Fas-associated phosphatase-1; Gene Activation; Gene Expression; Genes; Genetic Models; Glycosaminoglycans; Goals; Granulopoiesis; Growth Factor Receptors; Guanine Nucleotide Exchange Factors; HOX protein; HOXA10 gene; Hematopoietic; Human; ITGB3 gene; Immune response; Impairment; Innate Immune Response; Integrin alphaV; Karyotype; MLL gene; Mediating; Modeling; Molecular; Mus; Mutation; Myelopoiesis; Myeloproliferative disease; Ontology; Outcome; Oxidative Stress; PIK3CG gene; Pathway interactions; Patients; Phagocytes; Phosphorylation; Point Mutation; Process; Production; Progressive Disease; Protein Kinase; Protein phosphatase; Proteins; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Recurrent disease; Refractory Disease; Regulation; Relapse; Repression; Resistance; Role; Sampling; Signal Pathway; Signal Transduction; Stress; TP53 gene; Topoisomerase-II Inhibitor; Transcriptional Activation; acute myeloid leukemia cell; adverse outcome; aggressive therapy; autocrine; biological adaptation to stress; chemotherapy; chromosome loss; comorbidity; cytokine; design; drug relapse; homeodomain; immunoregulation; inhibitor/antagonist; knock-down; leukemia; leukemic stem cell; leukemogenesis; mouse model; neoplasm therapy; novel strategies; outcome forecast; overexpression; prevent; prognostic; protein expression; purine/pyrimidine metabolism; response; small molecule inhibitor; therapeutic target; transcription factor; transcriptome; translational approach; ubiquitin-protein ligase; wound

Acute myeloid leukemia (AML) is a heterogeneous disease with highly variable clinical outcomes. Recurring chromosomal abnormalities permit assignment of some AML patients to favorable vs unfavorable prognostic groups. Adverse prognosis is also associated with increased expression of a set of homeodomain transcription factors (HoxA7-11 and Meis1) and their target genes. This expression profile is found in AML with translocation or partial duplication of the MLL1 gene (i.e. 11q23-AML), translocations involving the MYST3 and CREBBP genes, and a subset of cytogenetically normal (CN) AML. We previously identified a set of common HoxA9/10 target genes that was enriched for cytokine receptors and their pathways, including Fgf2, Tgfβ2 and β3 integrin. HoxA9 and 10 cooperate to activate these genes in hematopoietic and leukemia stem cells (HSC and LSC). Consistent with this, we found that treatment with Fgf- R inhibitors (Fgf-R blocking antibody or the small molecule inhibitor, nintedanib) decreased proliferation and survival of CD34+ bone marrow cells from subjects with “Hox-profile” AML compared to samples from those without. In preliminary studies, we found that adding nintedanib to standard induction chemotherapy significantly prolonged remission and survival vs chemotherapy alone in a murine model of MLL1 rearranged leukemia. HoxA9 and A10 also regulate the innate immune response, but in this case their activities are antagonistic. During the innate immune response, activation of emergency granulopoiesis requires HoxA9, but HoxA10 is required to terminate the process. Termination required activation of the Triad1 gene by HoxA10; a process that involved overcoming repression by HoxA9. Triad1 is an E3 ubiquitin ligase that degrades growth factor receptors and Mdm2. We found Triad1 knockdown accelerated leukemogenesis in a murine model of 11q23-AML. In this murine model, we found the Lin-ckit+ LSC transcriptome was enriched for pathways involved in cytokine production, receptor tyrosine kinase (RTK) signaling, regulation of protein kinase activity, and positive regulation of the immune response vs control cells. We also found activation of pathways not previously associated with 11q23-AML, including Rap1 signaling. We profiled gene expression in mice in chemotherapy- induced remission (destined to relapse) vs chemotherapy + nintedanib (with sustained remission). We found differences in cytokine receptor activity, Pi3k/Akt signaling, guanine nucleotide exchange factor activity, purine/pyrimidine metabolism, oxidative stress response, and glycosamino-glycan biosynthesis. We hypothesize that adverse prognosis in Hox-overexpressing AML is characterized by impaired regulation of cytokine stimulated pathways and an activated stress response. These pathways represent rationale therapeutic targets to decrease chemotherapy resistance. This hypothesis will be pursued through three aims: Aim 1: Define the role of receptor tyrosine kinase pathways in drug resistance in Hox-overexpressing AML. We will study RTK/PI3K signaling pathways identified in our studies in bone marrow from murine AML models, including activity of anti-apoptotic BH3 proteins and protein phosphatase 2a (PP2a). Aim 2: Determine the impact of Rap1 signaling on disease progression in Hox-overexpressing AML. We will determine the role of various guanine nucleotide exchange factors on Rap or Ras activity, disease progression, and LSC persistence during chemotherapy. Aim 3: Investigate these pathways in AML patient samples. CD34+ bone marrow cells from AML patients at presentation or with relapsed or refractory disease after standard chemotherapy +/- approved agents will be analyzed. Hox-overexpressing AML will be compared to AML without increased Hox protein expression. The goal of these studies is to identify therapeutic targets to abolish or suppress LSCs during or after chemotherapy in adverse prognosis AML. This will be especially relevant to design new approaches to patients who are not candidates for aggressive treatment due to refractory disease or the presence of co-morbidities.

急性髓样白血病（AML）是一种异质性疾病，具有高度可变的临床结局。再次发生的 染色体异常允许某些AML患者分配有利与不利的预后 组。不良提示也与一组同源域转录的表达增加有关 因素（HOXA7-11和MEIS1）及其目标基因。此表达式曲线在AML中发现了易位 或MLL1基因的部分重复（即11q23-AML），涉及MYST3和CREBBP的易位 基因，以及细胞遗传学正常（CN）AML的子集。 我们先前鉴定了一组富含细胞因子接收器的常见HOXA9/10靶基因 以及它们的途径，包括FGF2，TGFβ2和β3整合素。 Hoxa9和10合作激活这些基因 造血和白血病干细胞（HSC和LSC）。与此相一致，我们发现用FGF-治疗 R抑制剂（FGF-R阻断抗体或小分子抑制剂Nintedanib）降低了增殖和 与这些样品相比 没有。在初步研究中，我们发现将Nyntedanib添加到标准诱导化疗中 在MLL1重新排列白血病的鼠模型中，仅长时间缓解和存活与化疗。 Hoxa9和A10还调节了先天免疫响应，但在这种情况下，它们的活动是对立的。 在先天免疫响应期间，紧急颗粒膜的激活需要Hoxa9，但Hoxa10是 终止过程所需。终止需要通过Hoxa10激活Triad1基因；一个过程 涉及hoxa9的克服代表。 Triad1是E3泛素连接酶，可降解生长因子受体 和MDM2。我们在11q23-AML的鼠模型中发现了Triad1敲低加速的白血病。 在此鼠模型中，我们发现Lin-Ckit+ LSC转录组丰富了涉及的途径 细胞因子产生，受体酪氨酸激酶（RTK）信号传导，蛋白激酶活性的调节和阳性 免疫反应与对照细胞的调节。我们还发现途径的激活 与11q23-AML相关，包括RAP1信号。我们在化学疗法中介绍了小鼠的基因表达 - 诱导的缓解（注定要缓解）与化学疗法 + Nyntedanib（持续缓解）。我们发现 细胞因子受体活性，PI3K/AKT信号传导，鸟嘌呤核丁基交换因子活性的差异， 嘌呤/嘧啶的代谢，氧化应激反应和糖尿氨基 - 聚糖生物合成。 我们假设在过表达HOX的AML中的不良预后的特征是调节受损 细胞因子刺激途径和激活的应力反应。这些途径代表理由 治疗靶标可降低化学疗法抗性。该假设将通过三个目标提出： AIM 1：定义受体酪氨酸激酶途径在过表达HOX的耐药性中的作用 AML。我们将研究在我们的研究中从Murine AML的骨髓研究中鉴定出的RTK/PI3K信号传导途径 模型，包括抗凋亡BH3蛋白和蛋白质磷酸酶2a（PP2A）的活性。 AIM 2：确定RAP1信号传导对过表达HOX的AML中疾病进展的影响。 我们将确定各种鸟嘌呤核丁基交换因子对RAP或RAS活性，疾病的作用 化疗期间的进展和LSC持久性。 AIM 3：在AML患者样品中研究这些途径。来自AML患者的CD34+骨髓细胞 标准化学疗法+/-批准的药物后，在介绍或中继或难治性疾病中将是 分析。将过表达HOX的AML与AML进行比较，而HOX蛋白表达不增加。 这些研究的目的是确定在或之后废除或抑制LSC的治疗靶点 提前化学疗法预后AML。这将与为患者设计新方法特别相关 由于难治性疾病或合并症的存在而不是候选人进行积极治疗的候选者。