Arsenic Trioxide and Acute Myeloid Leukemia

三氧化二砷与急性髓系白血病

• 批准号: 8034820
• 负责人: YONGKUI JING
• 金额: $ 24.74万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-15 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8034820
• 关键词: 4-ethoxymethylene-2-phenyl-2-oxazoline-5-one; Acids; Acute Myelocytic Leukemia; Acute Promyelocytic Leukemia; Aftercare; Animal Model; Apoptosis; Apoptotic; Arsenic Trioxide; Bacteria; Blood Cells; Blood specimen; Catabolism; Cell Death; Cell Line; Cells; Ceramides; Clinical; Clinical Trials; Combined Modality Therapy; Complex; Defect; Derivation procedure; Diuretics; Drug usage; Effectiveness; Enzymes; Esters; Ethacrynic Acid; Event; Fenretinide; Generations; Glutathione; Glutathione Reductase; Glutathione S-Transferase; Goals; HL60; Health; Human; Hydrogen Peroxide; Hydroxyl Radical; In complete remission; Individual; Induction of Apoptosis; K562 Cells; Leukemic Cell; Luciferases; MCL1 protein; Malignant Neoplasms; Mediating; Membrane; Messenger RNA; Mitochondria; Mus; NADPH Oxidase; NOD/SCID mouse; Organ; Pathway interactions; Patients; Peroxidases; Pharmaceutical Preparations; Physiological; Plasmids; Production; Proteins; Reactive Oxygen Species; Refractory; Relapse; Resistance; Respiration; Retinoids; Role; Sampling; Source; Sphingomyelins; Stratification; Superoxides; System; Tail; Testing; Therapeutic; Therapeutic Effect; Time; Toxic effect; Translating; Tretinoin; Tumor Burden; Weight; Xenograft procedure; analog; bioimaging; cancer cell; catalase; chemotherapy; clinical remission; design; efficacy testing; enzyme activity; glutathione peroxidase; in vivo; killings; leukemia; neutrophil; older patient; programs; response; small hairpin RNA; success

DESCRIPTION (provided by applicant): The outstanding therapeutic success of As2O3 in a subtype of acute myelocytic leukemia (AML), the acute promyelocytic leukemia (APL), prompted widespread effort to extend this therapy to other malignancies. We and others determined that, through generation of reactive oxygen species (ROS), As2O3 induced apoptosis of APL cells. We also showed that degradation of PML-RARa was not a crucial event in the pro-apoptotic program of As2O3, but high ROS production ability was. We thus postulate that by enhancing the ROS-generating capacity through combination therapies we will be able to effectively target non-APL AML for apoptosis. Membrane localized NADPH oxidase and mitochondria are the main sources of ROS production. AML cells share lineage derivation with polymorphonuclear cells, which use NADPH oxidase generated ROS to kill bacteria. We showed that As2O3 treatment of AML cells increases the expression, without causing activation, of a crucial component of NADPH oxidase, p47phox. Our preliminary results show that a combination of therapeutically achievable concentration of As2O3 with fenretinide (4-HPR), a retinoid approved for clinical trials, synergizes in causing apoptosis in AML cells which express detectable (basal or As2O3-induced) NADPH oxidase components and extends survival of AML bearing SCID/NOD mice. Under physiological conditions ROS generated as a byproduct of mitochondrial respiration are detoxified by glutathione-enzymes, glutathione peroxidase (GPx), glutathione-s-transferase ? (GST?) and glutathione reductase (GR). Inhibition of the activities of these enzymes combined with increased ROS generation due to malfunction of the mitochondrial respiration in cancer cells, should generate levels of ROS that are beyond tolerable, causing mitochondria-mediated apoptosis of AML cells. Indeed, a combination of As2O3, which inhibits GPx, with ethacrynic acid (a clinically used diuretic drug) and its new derivative, which we have developed, and which inhibit GST? and GR, synergistically induce apoptosis in AML cells. This effect is independent of NADPH oxidase expression. We propose to study the mechanisms underlying these findings and their effect in primary AML and in vivo (animal models) in 4 specific aims. In aim 1 we will study the mechanism of As2O3 induced NADPH oxidase expression and its activation by 4-HPR through a ceramide dependent pathway. In aim 2 we will study the mechanism of the synergy of apoptosis induction by As2O3 and ethacrynic acid, and its derivative, through inhibition of ROS degradation and decrease of an antiapoptotic protein Mcl-1. In aim 3 we will study the apoptotic effects of As2O3 and 4-HPR or As2O3 and ethacrynic acid in primary AML samples. Importantly, we will investigate the possibility of using NADPH oxidase/myeloperoxidase or GST?/catalase levels in primary human AML cells as potential predictive markers of patient stratification to sensitive or resistant to As2O3/4-HPR or As2O3/ethacrynic acid therapies, respectively. In specific aim 4 we will test the efficacy of the combination treatments in xenografts of human AML. If further studies confirm that these treatment combinations are indeed effective in inducing apoptosis in AML cell lines and primary cells, and that they have therapeutic effect in xenografts, without excessive toxicity, because of the current clinical use of each of the tested compounds, it will be possible to rapidly translate these finding into clinical trials. PUBLIC HEALTH RELEVANCE: Arsenic trioxide (As2O3) is a drug used world-wide that induces complete clinical remission in 90% of APL patients without significant toxicity. Our therapeutic design includes the use of 4 HPR, a drug already in clinical trial, that activates a membrane system for the production of ROS and the use of new non-diuretic derivatives of ethyacrynic acid which block enzymes that diminish ROS and markedly enhanceAs2O3 induced leukemic cell death by several mechanisms. Our goal will be to utilize As2O3 in combination with 4 HPR or EA analogs in clinical trials in patients with refractory, secondary or in elderly patients with acute myelogenous leukemia.

描述（由申请人提供）：As2O3在急性髓细胞白血病（AML）亚型，即急性早幼粒细胞白血病（APL）中的显著治疗成功，促使了将该疗法扩展到其他恶性肿瘤的广泛努力。我们和其他人确定，通过产生活性氧（ROS）， As2O3诱导APL细胞凋亡。我们还发现PML-RARa的降解不是As2O3促凋亡程序的关键事件，但高ROS生成能力是。因此，我们假设通过联合治疗增强ros生成能力，我们将能够有效地靶向非apl AML的细胞凋亡。膜定位NADPH氧化酶和线粒体是ROS产生的主要来源。AML细胞与多形核细胞共享谱系派生，多形核细胞利用NADPH氧化酶产生的ROS杀死细菌。我们发现，As2O3处理AML细胞增加了NADPH氧化酶的关键成分p47phox的表达，而不会引起激活。我们的初步结果表明，治疗上可达到的As2O3浓度与芬维甲酸（4-HPR）（一种批准用于临床试验的类维甲酸）的组合，可协同导致表达可检测的（基础或As2O3诱导的）NADPH氧化酶成分的AML细胞凋亡，并延长患有AML的SCID/NOD小鼠的生存期。在生理条件下，作为线粒体呼吸副产物产生的活性氧被谷胱甘肽酶、谷胱甘肽过氧化物酶（GPx）、谷胱甘肽s-转移酶？（GST）和谷胱甘肽还原酶（GR）抑制这些酶的活性，再加上癌细胞线粒体呼吸功能障碍导致ROS生成增加，应该会产生超出可耐受水平的ROS，导致线粒体介导的AML细胞凋亡。实际上，抑制GPx的As2O3与我们开发的乙稀酸（临床上使用的利尿剂）及其新衍生物的组合，抑制GST？和GR协同诱导AML细胞凋亡。这种作用不依赖于NADPH氧化酶的表达。我们拟在4个特定目的中研究这些发现的潜在机制及其在原发性AML和体内（动物模型）中的作用。在目的1中，我们将通过神经酰胺依赖途径研究As2O3诱导NADPH氧化酶表达及其被4-HPR激活的机制。在目的2中，我们将研究As2O3和乙酸及其衍生物通过抑制ROS降解和降低抗凋亡蛋白Mcl-1诱导细胞凋亡的协同作用机制。在目标3中，我们将研究As2O3和4-HPR或As2O3和乙酸在原发性AML样本中的凋亡作用。重要的是，我们将研究使用NADPH氧化酶/髓过氧化物酶或GST？/过氧化氢酶水平作为患者分层对As2O3/4- hpr或As2O3/乙酸治疗敏感或耐药的潜在预测标志物。在特定的目标4中，我们将测试联合治疗在人类AML异种移植中的疗效。如果进一步的研究证实这些治疗组合在诱导AML细胞系和原代细胞凋亡方面确实有效，并且它们对异种移植物有治疗效果，没有过度的毒性，因为目前每种被测试化合物的临床使用，将有可能迅速将这些发现转化为临床试验。公共卫生相关性：三氧化二砷（As2O3）是一种世界范围内使用的药物，可诱导90%的APL患者完全临床缓解，且无明显毒性。我们的治疗设计包括使用4 HPR，一种已经在临床试验中的药物，它可以激活一个膜系统来产生ROS，并使用新的非利尿剂乙酸衍生物，它可以阻断减少ROS的酶，并通过几种机制显著增强eas2o3诱导的白血病细胞死亡。我们的目标是将As2O3与4种HPR或EA类似物联合用于难治性、继发性或老年急性髓性白血病患者的临床试验。