Dissecting the Mitochondrial Translocation Mechanism of Anti-Apoptotic BCL-w

解析抗凋亡 BCL-w 的线粒体易位机制

• 批准号: 9192531
• 负责人: Edward Paul Harvey
• 金额: $ 4.36万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-04 至 2019-10-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9192531
• 关键词: Affinity Labels; Apoptosis; Apoptosis Regulation Gene; Apoptotic; BAX gene; BCL1 Oncogene; BCL2 gene; BH3 Domain; BH3 peptide; Binding; Biochemical; Biochemistry; Biological Assay; Biology; C-terminal; Cell Death; Cell Survival; Cells; Cellular translocation; Chemicals; Chemistry; Cytosol; Dana-Farber Cancer Institute; Development; Epitopes; Family; Goals; Homeostasis; Homologous Gene; Human; Induction of Apoptosis; Investigation; Lead; Length; Libraries; Ligands; Link; Maintenance; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Mediating; Medicine; Membrane; Methods; Mitochondria; Modeling; Molecular; Monitor; NMR Spectroscopy; Pathologic; Photoaffinity Labels; Physiological; Power Plants; Protein Family; Proteins; Regulation; Research Personnel; Research Proposals; Resistance; Series; Signal Transduction; Site; Stimulus; Stress; Structure; Time; Tissues; Training Programs; abstracting; affinity labeling; alpha helix; anticancer research; cancer cell; cytochrome c; design; human disease; innovation; medical schools; member; novel; novel therapeutic intervention; novel therapeutics; response; structural biology; treatment effect

PROJECT SUMMARY/ABSTRACT BCL-2 family proteins are key regulators of apoptosis, a form of programmed cell death essential to normal development and tissue homeostasis. Deregulation of the apoptotic machinery can lead to a host of human diseases characterized by too many or too few cells. Pathologic enforcement of cell survival by anti-apoptotic BCL-2 family proteins, which bind to and inactivate the BAX/BAK initiators of cell death, can lead to the development, maintenance, and chemoresistance of human cancer. In particular, anti-apoptotic BCL-w has been linked to the progression and invasiveness of a broad range of human cancers. BCL-w is believed to migrate from cytosol to mitochondria in response to interaction with BH3-only members of the BCL-2 family, but the physiological triggers and structure-function mechanism of BCL-w translocation are unknown. The Walensky lab has previously employed stapled “BH3” peptides, which recapitulate the natural structure and function of this critical signaling domain, to identify a novel regulatory site on pro-apoptotic BAX that mediates its mitochondrial translocation and resultant induction of apoptosis. Specifically, BH3 engagement of a groove formed by the juxtaposition of α-helices 1 and 6 of BAX induces allosteric release of its C-terminal helix for mitochondrial targeting and insertion. I hypothesize that BCL-w, as an anti-apoptotic homologue of BAX, may likewise be subject to regulation of its subcellular distribution. To elucidate the BCL-w translocation mechanism, I aim to: (1) synthesize and characterize stabilized alpha-helix of BCL-2 domains (SAHBs) modeled after BH3 helices to assess their functional interactions with anti-apoptotic BCL-w; (2) locate the binding interface and induced conformational changes associated with BH3-triggered BCL-w translocation; and (3) investigate the physiologic implications of stress-induced BCL-w translocation in cancer. Thus, the overarching goal of my proposal is to characterize the structure-function mechanism for ligand-stimulated BCL- w translocation and determine its contribution to enforcing apoptotic resistance in human cancer. I believe this study could inform a novel therapeutic strategy to disarm BCL-w in cancer by targeted disruption of the binding interface that drives its mitochondrial translocation. I am eager to embark on the rigorous training program proposed for my graduate studies at Harvard Medical School and the Dana-Farber Cancer Institute and look forward to developing as an independent and innovative investigator at the interface of chemical biology, apoptosis research, and cancer medicine.

项目摘要/摘要 Bcl-2家族蛋白是细胞凋亡的关键调节剂，这是一种正常的编程细胞死亡形式 发展和组织稳态。凋亡机制的放松管制会导致许多人 以太多或太少的细胞为特征的疾病。抗凋亡对细胞存活的病理执行 Bcl-2家族蛋白与细胞死亡的Bax/Bak启动者结合并失活，可能导致 人类癌症的开发，维持和化学抗性。特别是，抗凋亡BCl-W具有 我们与广泛的人类癌症的进步和侵入性联系在一起。据信BCL-W 响应与BCl-2家族的仅BH3成员的相互作用，从细胞质到线粒体迁移到线粒体 但是BCL-W易位的物理触发器和结构功能机理尚不清楚。这 Walensky Lab以前曾在“ BH3”肽中工作，该肽概括了自然结构和 这个关键信号域的功能，以识别培养基的促凋亡Bax上的新调节位点 它的线粒体易位并导致细胞凋亡。具体而言，BH3凹槽的参与度 由BAX的α-螺旋1和6的并置诱导其C末端螺旋的变构释放 线粒体靶向和插入。我假设Bcl-W是Bax的抗凋亡同源物，可能 同样，受其亚细胞分布的调节。阐明BCL-W易位 我的目标是：（1）合成并表征Bcl-2域（SAHB）的稳定α-螺旋（SAHB） 以BH3螺旋为模型，以评估其与抗凋亡Bcl-W的功能相互作用； （2）找到 与BH3触发的BCl-W易位相关的绑定界面和诱导的会议变化；和 （3）研究癌症中压力诱导的BCL-W易位的生理意义。那， 我的建议的总体目标是表征配体刺激的BCl-的结构功能机制 W易位并确定其对在人类癌症中强制凋亡抗性的贡献。我相信这个 研究可以通过针对结合的靶向破坏来告知一种新型的热策略，以解除癌症的BCL-W 驱动其线粒体易位的接口。我渴望开始严格的培训计划 我在哈佛医学院和达纳 - 弗伯癌研究所的研究生培训提议 在化学生物学界面上以独立和创新的研究者的形式发展， 凋亡研究和癌症医学。