Genetic Modifiers of the Anti-apoptotic Functions of Bcl-2

Bcl-2 抗凋亡功能的遗传修饰

• 批准号: 8003527
• 负责人: Cicely Anne Jette
• 金额: $ 9.38万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8003527
• 关键词: Antibodies; Apoptosis; Apoptotic; B-Cell Lymphomas; B-Lymphocytes; Biological Models; Cell Death; Cell Survival; Chimeric Proteins; Complement; DNA Damage; Development; Disease; Effectiveness; Embryo; Face; Follicular Lymphoma; Genes; Genetic; Goals; Hematologic Neoplasms; Hematological Disease; Hematopoietic Neoplasms; Human; Human Genome; Knowledge; Laboratories; Life Cycle Stages; Lymphoid Cell; Malignant Neoplasms; Maps; Mediating; Mentors; Modeling; Molecular; Mus; Mutate; Mutation; Normal Cell; Organ; Pathway interactions; Phenotype; Proteins; Radiation; Radiation Tolerance; Research; Resistance; Suppressor Mutations; Synteny; System; T-Lymphocyte; Tissues; Transgenic Organisms; Zebrafish; apoptosis in lymphoid cells; cancer cell; cancer therapy; design; in vivo; inhibitor/antagonist; mutant; novel; positional cloning; response; small molecule; thymocyte; tumor

This proposal is designed to provide the candidate a period of mentored research in the laboratory of Dr. A. T. Look, a pioneer in the field of hematopoietic malignancies and the use of zebrafish as a model system to study these diseases. The goal of this application is to use the zebrafish as a model to study novel genes and pathways necessary for the anti-apoptotic functions of Bcl-2, a gene whose aberrant expression is highly associated with follicular B-cell lymphoma and other hematological malignancies. The zebrafish, with its close synteny to the human genome and its conserved molecular pathways regulating the development of tissues and organs, offers a powerfultool with which to conduct such research. A transgenic zebrafish line that specifically expresses an EGFP-tagged zbcl-2 fusion protein in lymphoid cells has been employed in a genetic modifier screen, and the candidate has identified four zebrafish mutant lines that demonstrate suppression of the anti-apoptotic functions of Bcl-2 in vivo. The underlying hypothesis is that knowledge of the zebrafish genes able to suppress Bcl-2-mediated apoptosis in lymphoid cells with damaged DMAwill implicate novel pathways through which human BCL-2 exerts its anti-apoptotic activity in cancer cells. In Aim 1,the mutated genes will be mapped and positionally cloned. Mutations will be analyzed using a rag2- EGFP-mMyc zebrafish model of T-ALL to determine their ability to resensitize Bcl-2-expressing leukemic T cells to radiation-induced cell death. In Aim 2, functional and molecular analyses will be used to investigate the mechanisms by which each mutant gene suppresses the function of Bcl-2. The long-term goal of this application is to restore normal cell death pathways in B-cell follicular lymphoma and other hematological cancers by targeting pivotal molecules with small molecule inhibitors or antibodies. As part of their regulated life cycle, normal cells undergo programmed cell death (apoptosis) in response to DNA damage. Cancer cells derived from B-cell follicular lymphoma, as well as many other blood diseases, have aberrantly high levels of Bcl-2, a protein that causes cell survival in the face of apoptosis- inducing cancer therapies. The goal of this application is to use the zebrafish model system to identify targets for small molecule inhibitors of Bcl-2 function in order to restore normal cell death pathways in cancer cells.

该计划旨在为候选人提供一段时间的指导研究在博士的实验室。 a. t.听着，造血系统恶性肿瘤领域的先驱者和以斑马鱼为模型系统的研究者 来研究这些疾病。这项应用的目标是利用斑马鱼作为研究新基因的模型 以及Bcl-2的抗凋亡功能所必需的途径，Bcl-2是一种异常表达的基因， 与滤泡性B细胞淋巴瘤和其他血液恶性肿瘤相关。斑马鱼， 与人类基因组及其保守的调节发育的分子途径密切相关， 组织和器官，提供了一个强大的工具来进行这样的研究。转基因斑马鱼品系 在淋巴样细胞中特异性表达EGFP标记的zbcl-2融合蛋白的细胞已经被用于 遗传修饰剂筛选，候选人已经确定了四个斑马鱼突变系， 抑制Bcl-2在体内的抗凋亡功能。潜在的假设是， 斑马鱼基因能够抑制损伤DMA的淋巴细胞中Bcl-2介导的凋亡， 暗示了人BCL-2在癌细胞中发挥其抗凋亡活性新途径。在 目的1、对突变基因进行定位和克隆。突变将使用rag2- T-ALL的EGFP-mMyc斑马鱼模型，以确定其对表达Bcl-2的白血病T细胞再致敏的能力 辐射诱导的细胞死亡。在目标2中，将使用功能和分子分析来研究 每个突变基因抑制Bcl-2功能的机制。长期目标是 应用是恢复B细胞滤泡性淋巴瘤和其他血液系统疾病的正常细胞死亡途径。 通过用小分子抑制剂或抗体靶向关键分子来治疗癌症。 作为其调节的生命周期的一部分，正常细胞经历程序性细胞死亡（凋亡）以响应 DNA损伤。来自B细胞滤泡性淋巴瘤的癌细胞，以及许多其他血液 疾病，有异常高水平的Bcl-2，一种蛋白质，导致细胞存活在面对凋亡- 诱导癌症治疗。这个应用的目标是使用斑马鱼模型系统来识别 Bcl-2功能的小分子抑制剂的靶点，以恢复癌症中的正常细胞死亡途径 细胞