Redox Control of Apoptosis Signaling

细胞凋亡信号传导的氧化还原控制

• 批准号: 7625232
• 负责人: MARGARET M BRIEHL
• 金额: $ 22.13万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-15 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7625232
• 关键词: Antioxidants; Apoptosis; Attenuated; Biological Assay; Biological Models; CCAAT-Enhancer-Binding Protein-beta; CCAAT-Enhancer-Binding Proteins; Carrier Proteins; Cell Death; Cell Line; Cells; Complementary DNA; Data; Electron Transport; Environment; Enzymes; Fluorescent Probes; Gel; Generations; Genetic; Genetic Polymorphism; Glucocorticoid-induced apoptosis; Glucocorticoids; Glucosephosphate Dehydrogenase; Hexokinase 2; Hydrogen Peroxide; Immunoblotting; Immunohistochemistry; In Vitro; Knowledge; Lymphocyte; Lymphoma; Lymphoproliferative Disorders; Manganese Superoxide Dismutase; Metabolism; Mitochondria; Mitochondrial Proteins; Molecular Target; Mus; Nuclear Magnetic Resonance; Oxidation-Reduction; Oxidative Stress; Pharmaceutical Preparations; Phase; Production; Proteins; Proteomics; Reactive Oxygen Species; Research; Research Personnel; Resistance; Resistance development; Signal Transduction; Steroids; Superoxides; Testing; Thioredoxin; Thymic Lymphoma; Transfection; Transgenic Mice; Variant; Work; apoptosis in lymphocytes; base; cancer therapy; carbonyl group; catalase; chemotherapeutic agent; chemotherapy; clinically relevant; effective therapy; expectation; fatty acid oxidation; hexokinase; improved; inhibitor/antagonist; large cell Diffuse non-Hodgkin' s lymphoma; novel; overexpression; oxidation; programs; research study; resistance mechanism; response; transcription factor

DESCRIPTION (provided by applicant): The broad, long-term objective of the project is to elucidate mechanisms by which an altered cellular redox environment allows lymphoma cells to evade chemotherapy-induced apoptosis. Steroids are used as a first line therapy for lymphoma and development of resistance to these is a frequent barrier to effective treatment. The hypothesis to be tested is that mitochondria are key to the critical generation of reactive oxygen species during glucocorticoid-induced lymphocyte apoptosis and acquired tolerance to reactive oxygen species is a clinically-relevant mechanism of apoptosis resistance. The specific aims of the proposed research are as follows. 1) To use specific inhibitors of the electron transport chain and beta-oxidation of fatty acids, and cells depleted of functional mitochondria, to determine whether the reactive oxygen species which are critical for the signaling phase of glucocorticoid-induced apoptosis derive from mitochondria; and to use a novel fluorescent probe to test for differences in mitochondrial production of superoxide following glucocorticoid treatment of apoptosis-sensitive versus apoptosis-resistant cells. 2) To use immunoblots and 2D gels with proteomic analyses to identify key differences in mitochondrial proteins between glucocorticoid-sensitive and glucocorticoid-resistant lymphoma-derived cell lines; 2D gels will also be immunoblotted to detect significant differences in carbonyl groups that are generated as the result of protein oxidation. 3) To use immunohistochemistry and PCR-based assays to test whether decreased expression of manganese superoxide dismutase occurs in different types of lymphoma and is associated with polymorphisms that alter the transport of this protein into mitochondria or reduce its stability. 4) To use C/EBPbeta -/- transgenic mice to determine whether loss of this transcription factor in lymphocytes results in an altered cellular redox environment and resistance to apoptosis induced by chemotherapeutic agents used to treat lymphoma. Relevance: The proposed project is aimed at understanding why some lymphomas are not cured by current therapies. The experiments test the idea that treatment-resistant lymphoma cells have changes that make them resistant to oxidative stress and cell death (apoptosis). If this idea is correct, new, more effective anti-lymphoma drugs could be developed to counteract these changes.

描述（由申请人提供）：该项目的广泛，长期目标是阐明机制，通过这种机制，细胞氧化还原环境改变的机制使淋巴瘤细胞可以逃避化学疗法诱导的细胞凋亡。类固醇被用作淋巴瘤的第一线治疗，并且对这些耐药性的发展是有效治疗的常见障碍。要检验的假设是，线粒体是糖皮质激素诱导的淋巴细胞凋亡过程中活性氧的关键产生的关键，并且对活性氧的耐受性是一种临床上与凋亡抗性的临床抗性机制。拟议研究的具体目的如下。 1）使用电子传输链的特异性抑制剂以及脂肪酸的β-氧化以及功能性线粒体耗尽的细胞，以确定对糖皮质激素诱导的凋亡诱导的信号传导阶段至关重要的活性氧是否至关重要。并使用新型的荧光探针测试糖皮质激素治疗凋亡敏感性与抗凋亡耐药细胞后，超氧化物的线粒体产生差异。 2）使用蛋白质组学分析的免疫印迹和2D凝胶来鉴定糖皮质激素敏感和抗糖皮质激素耐药性淋巴瘤细胞系之间线粒体蛋白的关键差异；还将对2D凝胶进行免疫印迹，以检测因蛋白质氧化而产生的羰基的显着差异。 3）使用免疫组织化学和基于PCR的测定法测试锰超氧化物歧化酶的表达降低是否发生在不同类型的淋巴瘤中，并且与多态性有关，这些多态性会改变该蛋白质转移到线粒体中还是降低其稳定性。 4）使用C/EBPBETA - / - 转基因小鼠来确定淋巴细胞中这种转录因子的丧失是否会导致细胞氧化还原环境的改变，并且对治疗淋巴瘤的化学治疗剂诱导的凋亡抗性。相关性：拟议的项目旨在理解为什么某些淋巴瘤未通过当前疗法治愈。该实验测试了耐药性淋巴瘤细胞具有变化的观念，使它们抗氧化应激和细胞死亡（凋亡）。如果这个想法是正确的，可以开发出新的，更有效的抗淋巴瘤药物来抵消这些变化。