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: 9977962
• 负责人: Loren David Walensky
• 金额: $ 104.54万
• 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: 2015-08-10 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9977962
• 关键词: Address; Apoptosis; Apoptotic; BAX gene; BCL-2 Protein; BCL2 gene; BH4 Domain; Biochemistry; Biological; Biology; Cancer Research Project; Cell Death; Cells; Cellular biology; Cessation of life; Chemicals; Chemistry; Deuterium; Development; Equilibrium; Family; Family Research; Goals; Homeostasis; Homo; Human; Hydrogen; Laboratories; Laboratory Research; Life; Maintenance; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Membrane; Methods; Mitochondria; Molecular Conformation; Pediatric Oncologist; Pharmaceutical Preparations; Pharmacology; Protein Family; Proteins; Proteomics; Resistance; Signal Transduction; Site; Stress; Structure; Surface; Time; cancer cell; cancer therapy; inhibitor/antagonist; insight; interdisciplinary approach; mouse model; next generation; novel; novel strategies; novel therapeutic intervention; peptide structure; protein complex; public health relevance; response; small molecule; structural biology; tool

 DESCRIPTION (provided by applicant): The BCL-2 family of apoptotic proteins regulates the critical balance between cellular life and death. Deregulation of this essential signal transductio network drives the development, maintenance, and chemoresistance of a broad spectrum of human cancers. My laboratory is addressing the questions of just how interactions among BCL-2 family proteins regulate their structural changes and signaling functions during homeostasis and malignant transformation. Because BCL-2 protein interactions determine whether the cell will live or die in response to stress, we believe these structure-function studies will both infor basic biological mechanisms and opportunities to pharmacologically modulate them. Thirty years of BCL-2 family research has led to the first small molecule drug to reactive apoptosis in BCL-2 dependent cancer. Despite this remarkable scientific progress and the renewed promise of targeting BCL-2 proteins in cancer, fundamental questions remain about how this complex protein family regulates mitochondrial apoptosis. Indeed, their predominant membrane localization has made BCL-2 family proteins remarkably challenging to study. How the activated forms of BAX and BAK self-assemble into toxic oligomeric pores - the very death channels that mediate apoptosis - is unknown. How anti-apoptotic proteins such as BCL-2 change their structure within the membrane to block the various steps of BAX/BAK activation is unknown. Indeed, the full spectrum of contact surfaces between the many conserved domains of BCL-2 proteins is unknown. Perhaps the most perplexing question of how such structurally similar proteins can have diametrically opposite functions, as either inhibitors or activators of cell deat, remains essentially unknown. We address these mechanistic questions with broad experimental approaches, spanning chemistry, structural biology, proteomics, biochemistry, cell biology, mouse modeling, and pharmacology. For example, we have developed new chemical tools to dissect and target BCL-2 family protein interactions, advanced new methods to rapidly and accurately identify interaction sites using photoreactive structured peptides and mass spectrometry, and are currently applying hydrogen- deuterium exchange mass spectrometry to study BCL-2 family conformational changes in the membrane in real-time for the first time. Our goals for this R35 cancer research program include defining the conformational activation and homo-oligomerization mechanism(s) of BAX and BAK, characterizing a novel mechanism for BAX and BAK suppression by the BH4 domains of anti-apoptotic BCL-2 proteins, and investigating a new allosteric mechanism that controls the apoptotic functionalities of BCL-2 proteins. In each case, the structure- function insights will be harnessed to develop new approaches for targeting apoptotic resistance in cancer. As a chemical biologist and practicing pediatric oncologist, I have dedicated my research laboratory to deciphering BCL-2 family-mediated cancer mechanisms so that fresh insights into their protein interaction biology can inform the next generation of pro-apoptotic therapies for human cancer.

 描述（由适用提供）：凋亡蛋白的Bcl-2家族调节细胞生命与死亡之间的关键平衡。对这种基本信号转换网络的放松管制驱动了广泛的人类癌症的发育，维持和化学抗性。我的实验室正在解决Bcl-2家族蛋白之间的相互作用如何调节其结构变化和信号传导功能的问题。由于Bcl-2蛋白相互作用决定了细胞是否会响应压力而生命或死亡，因此我们认为这些结构功能研究将既融合了基本的生物学机制，又会发现药物调节它们的机会。 BCL-2家庭研究的30年导致了第一种小分子药物在Bcl-2依赖性癌症中的反应性凋亡。尽管这一显着的科学进步以及针对癌症中Bcl-2蛋白的新希望，但关于该复杂蛋白质家庭如何调节线粒体细胞凋亡的基本问题仍然是基本问题。实际上，它们主要的膜定位使Bcl-2家族蛋白蛋白质引起了极大的挑战。 Bax和Bak的激活形式如何自组装成有毒的寡聚孔（介导凋亡的死亡通道）是未知的。抗凋亡蛋白（例如Bcl-2）如何改变其在膜内的结构以阻断Bax/Bak激活的各个步骤。实际上，尚不清楚Bcl-2蛋白的许多配置结构域之间的全部接触表面。对于这种结构相似的蛋白质如何具有截然相反功能的最令人困惑的问题，例如细胞死亡的抑制剂或活化剂，基本上都是未知的。 We address these mechanical questions with broad experimental approaches, spanning chemistry, structural For example, we have developed new chemical tools to dissect and target BCL-2 family protein interactions, advanced new methods to rapidly and accurately identify interaction sites using photoreactive structured pepperides and mass spectrometry, and are currently applying hydrogen-deuterium exchange mass spectrometry to study BCL-2 family conformational changes in the membrane in real-time for the first time.我们的这项R35癌症研究计划的目标包括定义Bax和Bak的会议激活和同性化机制，表征了一种新型的BAX和BAK抑制的机制，并通过抗凋亡BCl-2蛋白的BH4域抑制了BH4域，并研究了一种新的变构机制，以控制BCL-2蛋白质的新型变构机制。在每种情况下，都将利用结构功能见解来开发针对癌症凋亡抗性的新方法。作为一名化学生物学家和练习小儿肿瘤学家，我专门研究了研究实验室对Bcl-2家族介导的癌症机制解密，以便对他们的蛋白质相互作用生物学的新见解可以告知下一代人类癌症的亲凋亡疗法。