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）：这项资助的授予的基本假设是，在用辐射或诱导DNA损伤的药物治疗后，可以抑制胸腺细胞和过表达BCL-2凋亡的基因的知识，将暗示相关，并导致BCL-2促进其抗磷脂的途径。作为原始父求目标的两个目标的一部分，进行了一个正向遗传筛选，并取得了出色的进步，以识别出在DNA损伤后抑制Bcl-2的新型突变体。在这些研究的过程中，提出了意外且令人兴奋的新发现，表明由内质网触发的较少研究的凋亡途径（ER）应力可以识别一组完全不同的靶标，可以介导对抗凋亡BCL-2途径的抑制。因此，我们正在提交这项竞争性修订应用程序，以增加新的研究目的，以利用有希望的ER应力诱导的凋亡途径作为一种机制，以使其敏感和杀死BCL-2过表达的癌细胞。值得注意的是，我们的初步研究表明，ER应力可以选择性地杀死斑马鱼幼虫中独立于Bcl-2状态的T细胞，并且我们已经具有令人信服的初步数据，该数据将斑马鱼确立为有力的新脊椎动物模型系统，用于剖析ER压力诱导的凋亡的分子机制。我们提出以下新目标3：识别和分析关键基因，这些基因在斑马鱼胚胎发生期间和T-All的斑马鱼模型中调节ER应力诱导的凋亡。由于我们在建立关键测定系统和试剂的初步研究方面取得了重大进展，因此我们“准备好铲”，通过在该ARRA刺激机制的一半和一半的时间内完成实验，从而在这一新目标上取得了重大进步。在实现这一目标时，将建立促凋亡BH3蛋白在ER应激中的胚胎作用，我们已经恢复了两个新型的ER应力增强子突变体，将在遗传上映射和分析，以及ER应力增强子突变对ER压力T-ALL细胞的影响。为了加速这些拟议的研究的进行，我们急需资金来保留乌贾瓦尔·帕蒂（Ujwal Pyati）博士，乌贾瓦尔·帕蒂（Ujwal Pyati）博士（目前支持2009年6月30日到期的NIH培训补助金），该资助于2009年6月30日到期，该补助金已经开发了我们对斑马AR应力模型的初步数据。我们还将立即聘请一名合格的研究技术专家，并订购新实验所需的耗材和试剂。这些实验将完成，描述这些结果的论文将在收到该竞争性更新申请支持的资金后的18个月内提交。 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.

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