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: 7811055
• 负责人: A. THOMAS LOOK
• 金额: $ 39万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7811055
• 关键词: Acute Lymphocytic Leukemia; Acute T Cell Leukemia; Aftercare; Animal Model; Animals; Antibodies; Apoptosis; Apoptotic; B-Lymphocytes; BCL2L11 gene; Biological Assay; Biological Models; Bypass; Caenorhabditis elegans; Cell Cycle Checkpoint; Cell Death; Cells; Cessation of life; Cloning; Coiled-Coil Domain; Complex; DNA Damage; Data; Defect; Development; Disease model; Effectiveness; Embryo; Embryonic Development; Enhancers; Environment; Ethylnitrosourea; Exhibits; Factor Analysis; Family; Family member; Fishes; Follicular Lymphoma; Funding; Funding Mechanisms; Genes; Genetic; Genetic Screening; Goals; Grant; Human; Human Resources; Hypoglycemia; Hypoxia; Knowledge; Laboratories; Larva; Leukemic Cell; Life; Malignant Neoplasms; Mammals; Maps; Mediating; Messenger RNA; Modeling; Molecular; Mouse-ear Cress; Mutagenesis; Mutate; Mutation; Nervous system structure; Normal Cell; Organism; Paper; Parents; Pathway interactions; Pattern; Pharmaceutical Preparations; Phenocopy; Phenotype; Point Mutation; Postdoctoral Fellow; Process; Protein Family; Proteins; Publications; Publishing; Puma; Qualifying; Radiation; Radiation Tolerance; Radiosensitization; Reagent; Reporting; Research; Resistance; Role; Saccharomyces cerevisiae; Screening procedure; Stimulus; Suppressor Mutations; System; T-Cell Leukemia; T-Lymphocyte; TP53 gene; Testing; Therapeutic; Therapeutic Agents; Time; Training; Translations; United States National Institutes of Health; Work; Yeasts; Zebrafish; alternative treatment; base; biological adaptation to stress; c-myc Genes; cancer cell; caspase-2; caspase-3; design; endoplasmic reticulum stress; falls; improved; in vivo; killings; leukemia; mutant; novel; overexpression; parent grant; public health relevance; research study; response; small molecule; thymocyte; tool; tumor

DESCRIPTION (provided by applicant): The underlying hypothesis of this funded grant is that knowledge of the genes that can suppress apoptosis in thymocytes and cancer cells overexpressing BCL-2, after treatment with radiation or drugs that induce DNA damage, will implicate relevant, and drug targetable, pathways through which BCL-2 exerts its anti-apoptotic activity. As part of the two aims of the original parent proposal, a forward genetic screen was conducted and outstanding progress has been made identifying novel mutants that suppress BCL-2 following DNA damage. During the course of these studies, unexpected and exciting new findings were made indicating that a less studied apoptotic pathway triggered by Endoplasmic Reticulum (ER) stress can identify a completely different set of targets that can mediate suppression of the anti-apoptotic BCL-2 pathway. Thus, we are submitting this competitive revision application to add a new research aim for our grant to exploit the promising ER stress-induced apoptotic pathway as a mechanism to sensitize and kill BCL-2-overexpressing cancer cells. Remarkably, our preliminary studies show that ER stress can selectively kill T-cells in zebrafish larvae independent of BCL-2 status and we already have convincing preliminary data that establishes the zebrafish as a powerful new vertebrate model system for dissecting the molecular mechanisms of ER stress-induced apoptosis. We propose the following New Aim 3: To identify and analyze critical genes that modulate ER stress-induced apoptosis in the context of overexpressed BCL-2 during zebrafish embryogenesis and in zebrafish models of T-ALL. Due to our significant progress in preliminary studies establishing critical assay systems and reagents, we are 'shovel ready' to make major advances on this new Aim through experiments that can definitely be completed within the one and one-half year time-frame of this ARRA stimulus mechanism. In accomplishing this aim, the embryonic role of pro-apoptotic BH3-only proteins in ER stress will be established, two novel ER stress enhancer mutants that we already have recovered will be genetically mapped and analyzed, and the effects of ER stress enhancer mutations on ER-stressed T-ALL cells will be determined. In order to accelerate the conduct of these proposed studies we urgently need funding to retain Dr. Ujwal Pyati, a postdoctoral fellow currently supported on an NIH training grant that expires June 30, 2009, who developed our preliminary data on the zebrafish ER stress model. We also will immediately hire a qualified research technologist and order the supplies and reagents needed for our new experiments. These experiments will be completed and the papers describing these results will be submitted within 18 months of receipt of the funds supported by this competitive renewal application. PUBLIC HEALTH RELEVANCE: The discovery of BCL-2 suppressor mutations will identify pivotal targets whose inhibition by antibodies or small molecules will restore normal cell death pathways, thus enhancing the effectiveness of apoptosis-inducing therapies for B-cell follicular lymphoma and other BCL-2-overexpressing apoptosis-resistant cancers in humans.

描述（由申请人提供）：这项资助的基本假设是，在用辐射或诱导 DNA 损伤的药物治疗后，了解可以抑制胸腺细胞和过度表达 BCL-2 的癌细胞凋亡的基因，将暗示 BCL-2 发挥其抗凋亡活性的相关且药物可靶向的途径。作为最初母提案的两个目标的一部分，进行了正向遗传筛选，并在识别 DNA 损伤后抑制 BCL-2 的新型突变体方面取得了显着进展。在这些研究过程中，出现了意想不到且令人兴奋的新发现，表明由内质网 (ER) 应激触发的较少研究的细胞凋亡途径可以识别一组完全不同的靶标，这些靶标可以介导抗凋亡 BCL-2 途径的抑制。因此，我们提交这份竞争性修订申请，为我们的资助增加一个新的研究目标，即利用有前途的 ER 应激诱导细胞凋亡途径作为敏化和杀死 BCL-2 过表达癌细胞的机制。值得注意的是，我们的初步研究表明，ER 应激可以选择性地杀死斑马鱼幼虫中的 T 细胞，与 BCL-2 状态无关，并且我们已经有了令人信服的初步数据，将斑马鱼确立为一个强大的新脊椎动物模型系统，用于剖析 ER 应激诱导细胞凋亡的分子机制。我们提出以下新目标 3：识别和分析在斑马鱼胚胎发生期间和 T-ALL 斑马鱼模型中过表达 BCL-2 的情况下调节 ER 应激诱导细胞凋亡的关键基因。由于我们在建立关键检测系统和试剂的初步研究方面取得了重大进展，我们已做好准备，通过实验在这一新目标上取得重大进展，这些实验绝对可以在 ARRA 刺激机制的一年半时间内完成。为了实现这一目标，将确定促凋亡 BH3-only 蛋白在 ER 应激中的胚胎作用，对我们已经恢复的两种新型 ER 应激增强子突变体进行基因图谱和分析，并确定 ER 应激增强子突变对 ER 应激 T-ALL 细胞的影响。为了加速这些拟议研究的进行，我们迫切需要资金来留住 Ujwal Pyati 博士，他是一名博士后研究员，目前接受 NIH 培训补助金的支持，该补助金将于 2009 年 6 月 30 日到期，他为我们开发了斑马鱼 ER 压力模型的初步数据。我们还将立即聘请一名合格的研究技术专家，并订购我们新实验所需的用品和试剂。这些实验将在收到此竞争性续订申请支持的资金后 18 个月内完成，并提交描述这些结果的论文。 公共健康相关性：BCL-2抑制突变的发现将确定关键靶标，通过抗体或小分子抑制这些靶标将恢复正常的细胞死亡途径，从而增强细胞凋亡诱导疗法对人类B细胞滤泡性淋巴瘤和其他BCL-2过表达的抗凋亡癌症的有效性。

项目成果

p63 mediates an apoptotic response to pharmacological and disease-related ER stress in the developing epidermis.

• DOI: 10.1016/j.devcel.2011.07.012
• 发表时间: 2011-09-13
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Pyati, Ujwal J.;Gjini, Evisa;Carbonneau, Seth;Lee, Jeong-Soo;Guo, Feng;Jette, Cicely A.;Kelsell, David P.;Look, A. Thomas
• 通讯作者: Look, A. Thomas