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

这项建议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.Look，血液病领域的先驱，并使用斑马鱼作为模型系统 来研究这些疾病。这项应用的目标是以斑马鱼为模型来研究新的基因。 以及抗凋亡功能所必需的通路，这是一种高度异常表达的基因 与滤泡性B细胞淋巴瘤和其他血液系统恶性肿瘤有关。斑马鱼，和它的 与人类基因组及其调控发育的保守分子通路密切相关 组织和器官，为进行此类研究提供了一个强有力的工具。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融合蛋白的基因已被用于 遗传修饰物筛选，候选人已经确定了四个斑马鱼突变系，这些突变系证明了 体内抑制Bcl2的抗细胞凋亡功能。潜在的假设是知道 斑马鱼基因可抑制DNA损伤的淋巴样细胞中Bcl2介导的细胞凋亡 揭示人类bcl2在癌细胞中发挥抗凋亡活性的新途径。在……里面 目的1、对突变基因进行定位和克隆。突变将使用RAG2- EGFP-mMyc斑马鱼T-ALL模型检测其对表达Bcl2的白血病T细胞的再增敏能力 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Bc l-2功能的机制。这样做的长期目标是 应用于恢复B细胞滤泡性淋巴瘤和其他血液病的正常细胞死亡途径 通过使用小分子抑制剂或抗体靶向关键分子来治疗癌症。 作为其调节生命周期的一部分，正常细胞经历程序性细胞死亡(凋亡)作为回应 造成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，这是一种在面临凋亡的情况下导致细胞存活的蛋白质- 诱导癌症治疗。这个应用程序的目标是使用斑马鱼模型系统来识别 以小分子抑制物为靶点恢复肿瘤细胞的正常死亡途径 细胞。

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.