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: 7081687
• 负责人: Cicely Anne Jette
• 金额: $ 13.28万
• 依托单位: DANA-FARBER CANCER INST
• 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/7081687
• 关键词: B cell lymphoma; BCL2 gene /protein; DNA damage; T lymphocyte; apoptosis; autosomal recessive trait; gamma radiation; gene complementation; gene expression; gene induction /repression; gene mutation; genetic mapping; molecular genetics; neoplasm /cancer genetics; p53 gene /protein; positional cloning; site directed mutagenesis; small molecule; zebrafish

DESCRIPTION (provided by applicant): 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 powerful tool 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 DNA will 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.

描述（由申请人提供）： 该计划旨在为候选人提供一段在A博士实验室进行指导研究的时间。T.他是造血系统恶性肿瘤研究领域的先驱，并将斑马鱼作为研究这些疾病的模型系统。本申请的目的是使用斑马鱼作为模型来研究Bcl-2的抗凋亡功能所必需的新基因和途径，Bcl-2是一种异常表达与滤泡性B细胞淋巴瘤和其他血液恶性肿瘤高度相关的基因。斑马鱼与人类基因组具有密切的同线性，其保守的分子途径调节组织和器官的发育，为进行此类研究提供了强有力的工具。在淋巴样细胞中特异性表达EGFP标记的zbcl-2融合蛋白的转基因斑马鱼系已被用于遗传修饰剂筛选，并且候选人已鉴定出四种斑马鱼突变系，其证明在体内抑制Bcl-2的抗凋亡功能。潜在的假设是，斑马鱼基因能够抑制Bcl-2介导的细胞凋亡的淋巴细胞与受损的DNA的知识将牵连新的途径，通过人BCL-2发挥其抗凋亡活性的癌细胞。在目标1中，突变基因将被定位和克隆。将使用T-ALL的rag 2-EGFP-mMyc斑马鱼模型分析突变，以确定其使表达Bcl-2的白血病T细胞对辐射诱导的细胞死亡再敏感的能力。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。本申请的长期目标是通过用小分子抑制剂或抗体靶向关键分子来恢复B细胞滤泡性淋巴瘤和其他血液癌症中的正常细胞死亡途径。 作为其调节的生命周期的一部分，正常细胞响应于DNA损伤而经历程序性细胞死亡（凋亡）。来自B细胞滤泡性淋巴瘤以及许多其他血液疾病的癌细胞具有异常高的Bcl-2水平，Bcl-2是一种蛋白质，在面对诱导凋亡的癌症治疗时会导致细胞存活。该应用的目标是使用斑马鱼模型系统来识别Bcl-2功能小分子抑制剂的靶点，以恢复癌细胞的正常细胞死亡途径。