Dissecting and Targeting Deregulated Mitochondrial Apoptosis in Human Cancer

剖析和靶向人类癌症中失调的线粒体凋亡

• 批准号: 10299794
• 负责人: Loren David Walensky
• 金额: $ 107.18万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-24 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10299794
• 关键词: Advanced Development; Apoptosis; Apoptotic; BAX gene; BCL-2 Protein; BCL2 gene; Binding; Binding Sites; Biochemistry; Cancer Biology; Cells; Cellular biology; Cessation of life; Chemicals; Chemistry; Chemoresistance; Cysteine; Development; Enzymes; Family; Goals; Human; Length; Ligands; Long-Chain-Acyl-CoA Dehydrogenase; MCL1 gene; Maintenance; Malignant Neoplasms; Mediating; Metabolic; Mitochondria; Modification; Molecular; Molecular Conformation; Pathogenesis; Pathologic; Pathway interactions; Pediatric Oncologist; Pharmacology; Phase; Post-Translational Protein Processing; Protein Family; Proteins; Proteomics; Regulation; Relapse; Research; Resistance; Role; Signal Pathway; Structure; Surface; Technology; Work; apoptosis deregulation; cancer therapy; fatty acid metabolism; fatty acid oxidation; in vivo evaluation; inhibitor/antagonist; insight; interdisciplinary approach; monomer; mutant; next generation; novel; novel therapeutics; programs; prototype; refractory cancer; structural biology; tool; tumor metabolism

PROJECT SUMMARY/ABSTRACT BCL-2 proteins participate in a dynamic interaction network that determines whether a cell will live or die. Deregulation of this essential signaling pathway underlies the pathogenesis of human cancer and resistance to treatment. The goal of this R35 research program is to elucidate the fundamental protein interaction mechanisms that drive the apoptotic program and harness these insights to develop next-generation cancer treatments. Over the last five years of R35 support, we applied novel chemical tools and a host of analytical technologies to achieve mechanistic discoveries that revealed new druggable binding sites and compounds to target them. We found that covalent modification of distinct cysteines in pro-apoptotic BAX and anti-apoptotic MCL-1 and BFL-1 differentially regulate their apoptotic functions. Our pursuit of covalent ligands that mimic these post-translational modifications are yielding prototype BAX activators and MCL-1 and BFL-1 inhibitors for cancer therapy. Deciphering how BAX and BAK are directly activated, and the conformational mechanisms that underlie their conversion from latent monomers into toxic mitochondrial oligomers, has also been a major focus of our work. Indeed, the elusive structures of the BAX and BAK death channels represent the “holy grail” of apoptosis research. We recently generated the first full-length homogeneous BAX oligomer (BAXO) amenable to structure- function characterizations, providing a glimpse into the macromolecular organization of a functional BAXO species. BAXO and its mutants are enabling us to pinpoint the structural determinants for each step of the BAX- activation pathway and thus inform new control points for pharmacologic activation of apoptosis. In addition to dissecting these high-priority, canonical BCL-2 protein interactions, we have developed proteomic tools to identify non-canonical targets and recently found that MCL-1 directly interacts with the fatty acid oxidation enzyme VLCAD, revealing a dual role for MCL-1 at the intersection of apoptosis and metabolic regulation. We hypothesize that MCL-1-driven cancers rely on both apoptotic suppression and fatty acid metabolism to maximize pathologic survival, potentially explaining why MCL-1 is the most widely expressed anti-apoptotic protein across human cancers. Here, we build on our newest mechanistic insights to interrogate a spectrum of BCL-2 family interactions that drive human cancer and mine each opportunity to pharmacologically subvert them. Specifically, our next set of R35 goals are: (1) identify the structural and functional determinants that mediate the “execution phase” of mitochondrial apoptosis; (2) solve the structure of a BAX oligomer; (3) characterize the non-canonical role of MCL-1 at the intersection of apoptosis and cancer metabolism; and (4) advance the development and in vivo testing of BCL-2 family molecular modulators as next-generation therapies for human cancer. We tackle these goals using multidisciplinary approaches that span chemistry, structural biology, proteomics, biochemistry, cell biology, and in vivo testing. As a chemical biologist and pediatric oncologist, I am committed to transforming our fresh mechanistic insights into new therapies for relapsed and refractory cancers.

项目摘要/摘要 Bcl-2蛋白参与一个动态交互网络，该网络确定单元格是生命还是死亡。 这种埃经信号通路的放松管制是人类癌症的发病机理和对 治疗R35研究计划的目的是阐明基本蛋白质相互作用 这推动了凋亡计划，并利用这些见解发展下一代癌症 R35支持的最后五年，我们应用了新颖的化学工具和许多分析技术 实现启示新的可药物结合的机理发现，并瞄准了我们 发现在促凋亡的Bax抗磷脂MCL-1和BFL-1中，不同半胱氨酸的共价修饰 差异调节其凋亡功能。 修改是屈服原型Bax激活剂，MCL-1和BFL-1抑制剂进行癌症治疗。 破译Bax和Bak是如何直接激活的，以及其基础的同意机制 从后期的单体转换为tomitochrial寡聚物，也是我们工作的重点。 确实，Bax和Bak死亡通道的难以捉摸的结构代表了凋亡的“圣杯” 研究。 功能表征，可瞥见功能背部的大分子组织 物种。 激活途径，从而为凋亡的药理学激活提供了新的控制点 这些高优先级，规范的Bcl-2蛋白相互作用，我们已经开发了蛋白质组学 识别非经典靶标，最近发现MCL-1与脂肪酸直接相互作用 酶VLCAD，在凋亡和代谢调节的交集中，MCL-1的双重作用 假设MCL-1驱动的癌症既依赖于示例性抑制和脂肪酸代谢 最大化病理生存，有可能解释为什么MCL-1是最广泛表达的抗凋亡 遍布人类癌的蛋白质。 Bcl-2家庭互动，这些家庭互动会推动人类癌症，并为药理学颠覆它们的每一个机会。 具体而言，我们的下一组R35目标是：（1）确定结构和功能决定因素的结构和功能决定因素 Mitchondrial凋亡的“执行阶段”； MCL-1在凋亡和癌症代谢的交点中的非传统作用； Bcl-2家族分子调节剂的开发和体内测试作为人类的下一代疗法 癌症。 蛋白质组学，生物化学，细胞生物学和体内测试。 致力于将我们的新机械洞察力转变为新的疗法，以恢复难治性癌症。