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: 10265363
• 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/10265363
• 关键词: 11q23; Acute Myelocytic Leukemia; Alkylating Agents; Anabolism; Apoptotic; Blocking Antibodies; Bone Marrow; Bone Marrow Cells; CD34 gene; CREBBP gene; Cells; Chemoresistance; 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; 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; Prognosis; Progressive Disease; Protein Kinase; Protein phosphatase; Proteins; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Recurrent disease; Refractory Disease; Regulation; Relapse; Repression; 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; 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.

急性髓系白血病（AML）是一种异质性疾病，临床结局高度可变。经常性 染色体异常允许将一些AML患者分配为有利与不利的预后 组不良预后也与一组同源域转录的表达增加有关。 因子（HoxA 7 -11和Meis 1）及其靶基因。这种表达谱见于AML易位 或MLL 1基因的部分重复（即11 q23-AML），涉及MYST 3和CREBBP的易位 基因，和细胞遗传学正常（CN）AML的子集。 我们先前鉴定了一组常见的HoxA 9/10靶基因，其富含细胞因子受体， 及其途径，包括Fgf 2、TGFβ2和β3整合素。HoxA 9和10合作激活这些基因， 造血干细胞和白血病干细胞（HSC和LSC）。与此一致，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尼达尼布）可降低增殖， 来自“Hox-特征”AML受试者的CD 34+骨髓细胞与来自“Hox-特征”AML受试者的样本相比的存活率 没有.在初步研究中，我们发现在标准诱导化疗中添加尼达尼布， 在MLL 1重排白血病小鼠模型中，与单独化疗相比，缓解期和生存期延长。 HoxA 9和A10也调节先天免疫应答，但在这种情况下，它们的活性是拮抗的。 在先天性免疫应答期间，紧急粒细胞生成的激活需要HoxA 9，但HoxA 10是 需要终止进程。终止需要通过HoxA 10激活Triad 1基因;这一过程 包括克服HoxA 9的抑制Triad 1是一种E3泛素连接酶，可降解生长因子受体 MDM2。我们发现Triad 1基因敲低可加速11 q23-AML小鼠模型的白血病发生。 在这个小鼠模型中，我们发现Lin-ckit+ LSC转录组富集了参与以下过程的途径： 细胞因子的产生、受体酪氨酸激酶（RTK）信号传导、蛋白激酶活性的调节，以及阳性 与对照细胞相比，免疫应答的调节。我们还发现了以前没有的通路的激活 与11 q23-AML相关，包括Rap 1信号。我们分析了化疗小鼠的基因表达- 诱导缓解（注定复发）vs化疗+尼达尼布（持续缓解）。我们发现 细胞因子受体活性，Pi 3 k/Akt信号传导，鸟嘌呤核苷酸交换因子活性， 嘌呤/嘧啶代谢、氧化应激反应和糖胺聚糖生物合成。 我们假设Hox过度表达AML的不良预后以调节受损为特征， 细胞因子刺激的途径和激活的应激反应。这些途径代表了 降低化疗耐药性的治疗靶点。这一假设将通过三个目标来实现： 目的1：明确受体酪氨酸激酶通路在Hox过表达的耐药细胞中的作用。 急性髓细胞白血病我们将研究在我们的研究中鉴定的小鼠AML骨髓中的RTK/PI 3 K信号通路 模型，包括抗凋亡BH 3蛋白和蛋白磷酸酶2a（PP 2a）的活性。 目的2：确定Rap 1信号对Hox过表达AML疾病进展的影响。 我们将确定各种鸟嘌呤核苷酸交换因子对Rap或Ras活性的作用， 进展和化疗期间LSC持续性。 目的3：研究AML患者样本中的这些途径。来自AML患者的CD 34+骨髓细胞 在标准化疗+/-批准的药物治疗后， 分析了将Hox过表达AML与Hox蛋白表达不增加的AML进行比较。 这些研究的目的是确定治疗靶点，以消除或抑制LSC期间或之后， 化疗治疗预后不良的急性髓细胞白血病。这将是特别相关的设计新的方法，病人 由于难治性疾病或合并症的存在而不适合积极治疗的患者。