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

基本信息

批准号：
8955883
负责人：
Loren David Walensky
金额：
$ 104.34万
依托单位：
DANA-FARBER CANCER INST
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-10 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8955883
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 Complex 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 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 therapeutics peptide structure public health relevance response small molecule structural biology tool

项目摘要

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（由申请人提供）：BCL-2 凋亡蛋白家族调节细胞生命与死亡之间的关键平衡，这种重要信号转导网络的失调会驱动多种人类癌症的发展、维持和化学耐药性。正在解决 BCL-2 家族蛋白之间的相互作用如何在稳态和恶性转化过程中调节其结构变化和信号传导功能的问题，因为 BCL-2 蛋白相互作用决定细胞的生存或死亡。三十年的 BCL-2 家族研究已经开发出了第一种能够对 BCL-2 依赖性癌症进行反应性细胞凋亡的小分子药物。尽管取得了显着的科学进展，并且在癌症中靶向 BCL-2 蛋白有了新的希望，但关于这个复杂的蛋白家族如何调节线粒体凋亡的基本问题仍然存在，事实上，它们的主要膜定位使得 BCL-2 家族蛋白的研究变得非常具有挑战性。激活的形式BAX 和 BAK 自组装成有毒寡聚孔（介导细胞凋亡的死亡通道）尚不清楚 BCL-2 等抗细胞凋亡蛋白如何改变其膜内结构以阻止 BAX/BAK 激活的各个步骤。事实上，BCL-2 蛋白的许多保守结构域之间的接触面的全部范围尚不清楚，也许最令人困惑的问题是，这种结构相似的蛋白如何具有截然相反的功能，无论是作为抑制剂还是作为抑制剂。我们通过广泛的实验方法来解决这些机制问题，涵盖化学、结构生物学、蛋白质组学、生物化学、细胞生物学、小鼠模型和药理学。针对BCL-2家族蛋白相互作用，先进的新方法利用光反应结构肽和质谱快速准确地识别相互作用位点，目前正在应用氢-氘交换质谱来研究BCL-2家族我们的 R35 癌症研究计划的目标是首次实时检测膜中的构象变化，包括定义 BAX 和 BAK 的构象激活和同源寡聚机制，描述通过抑制 BAX 和 BAK 的新机制。抗凋亡 BCL-2 蛋白的 BH4 结构域，并研究控制 BCL-2 蛋白凋亡功能的新变构机制。作为一名化学生物学家和执业儿科肿瘤学家，我致力于开发针对癌症细胞凋亡抵抗的新方法，我的研究实验室致力于破译 BCL-2 家族介导的癌症机制，以便对其蛋白质相互作用生物学的新见解可以为下一代提供信息。人类癌症的促凋亡疗法。