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：
10669117
负责人：
Loren David Walensky
金额：
$ 103.12万
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：
2015-08-24 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10669117
关键词：
Advanced Development Apoptosis Apoptotic BAX gene BCL-2 Protein BCL1 Oncogene 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 Permeability Phase Post-Translational Protein Processing Protein Family Proteins Proteomics Regulation Relapse Research Resistance Role Signal Pathway Structure Surface Technology Work cancer therapy fatty acid metabolism fatty acid oxidation in vivo evaluation inhibitor insight interdisciplinary approach monomer mutant next generation novel novel therapeutics pharmacologic programs prototype refractory cancer structural biology structural determinants tool tumor metabolism

项目摘要

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.

项目概要/摘要 BCL-2 蛋白参与动态相互作用网络，决定细胞的生存或死亡。这一重要信号通路的失调是人类癌症的发病机制和耐药性的基础该 R35 研究计划的目标是阐明基本的蛋白质相互作用机制。驱动细胞凋亡程序并利用这些见解来开发下一代癌症治疗方法。在 R35 支持的最后五年中，我们应用了新颖的化学工具和大量分析技术实现机制发现，揭示新的可成药结合位点和针对它们的化合物。发现促凋亡 BAX 和抗凋亡 MCL-1 和 BFL-1 中不同半胱氨酸的共价修饰我们寻找模仿这些翻译后功能的共价配体。修饰正在产生用于癌症治疗的 BAX 激活剂原型以及 MCL-1 和 BFL-1 抑制剂。破译 BAX 和 BAK 如何直接激活，以及它们背后的构象机制从潜在单体转化为有毒线粒体寡聚体也是我们工作的一个主要焦点。事实上，BAX 和 BAK 死亡通道的难以捉摸的结构代表了细胞凋亡的“圣杯” 我们最近生成了第一个全长均质 BAX 寡聚物（BAXO），适合结构- 功能表征，让您一睹功能性 BAXO 的大分子组织 BAXO 及其突变体使我们能够查明 BAX-每一步的结构决定因素。激活途径，从而为细胞凋亡的药理学激活提供新的控制点。通过剖析这些高优先级、典型的 BCL-2 蛋白质相互作用，我们开发了蛋白质组学工具识别非典型靶标，最近发现 MCL-1 直接与脂肪酸氧化相互作用 VLCAD 酶，揭示了 MCL-1 在细胞凋亡和代谢调节交叉点上的双重作用。认为 MCL-1 驱动的癌症依赖于细胞凋亡抑制和脂肪酸代谢最大限度地提高病理存活率，这可能解释了为什么 MCL-1 是表达最广泛的抗凋亡药物在这里，我们基于最新的机制见解来探究一系列的蛋白质。 BCL-2 家族的相互作用会导致人类癌症，并挖掘每一个机会从药理上颠覆它们。具体来说，我们的下一组 R35 目标是：(1) 确定介导的结构和功能决定因素线粒体凋亡的“执行阶段”；(2) 解析 BAX 寡聚体的结构；(3) 表征 MCL-1 在细胞凋亡和癌症代谢交叉点中的非典型作用；(4) 促进 BCL-2 家族分子调节剂作为人类下一代疗法的开发和体内测试我们使用跨化学、结构生物学、癌症等多学科方法来实现这些目标。作为一名化学生物学家和儿科肿瘤学家，我是蛋白质组学、生物化学、细胞生物学和体内测试。致力于将我们新的机制见解转化为复发性和难治性癌症的新疗法。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Site-Dependent Cysteine Lipidation Potentiates the Activation of Proapoptotic BAX.

位点依赖性半胱氨酸脂化增强促凋亡 BAX 的激活。

DOI：
发表时间：
2020
期刊：
影响因子：
8.8
作者：
Cohen, Daniel T;Wales, Thomas E;McHenry, Matthew W;Engen, John R;Walensky, Loren D
通讯作者：
Walensky, Loren D

Identification of a Structural Determinant for Selective Targeting of HDMX.

HDMX 选择性靶向的结构决定因素的鉴定。

DOI：
10.1016/j.str.2020.04.011
发表时间：
2020-04-29
期刊：
Structure
影响因子：
5.7
作者：
Y. Ben;H. Seo;Edward P. Harvey;Zachary J. Hauseman;T. Wales;C. Newman;A. Cathcart;J. Engen;S. Dhe;L. Walensky
通讯作者：
L. Walensky

Allosteric sensitization of proapoptotic BAX.

促凋亡 BAX 的变构敏化。

DOI：
发表时间：
2017-09
期刊：
影响因子：
14.8
作者：
Pritz, Jonathan R;Wachter, Franziska;Lee, Susan;Luccarelli, James;Wales, Thomas E;Cohen, Daniel T;Coote, Paul;Heffron, Gregory J;Engen, John R;Massefski, Walter;Walensky, Loren D
通讯作者：
Walensky, Loren D

Targeting BAX to drug death directly.

直接针对 BAX 进行药物死亡。

DOI：
发表时间：
2019
期刊：
影响因子：
14.8
作者：
Walensky; Loren D
通讯作者：
Loren D

Design of stapled antimicrobial peptides that are stable, nontoxic and kill antibiotic-resistant bacteria in mice.

设计稳定、无毒并能杀死小鼠体内抗生素耐药细菌的钉合抗菌肽。

DOI：
发表时间：
2019
期刊：
影响因子：
46.9
作者：
Mourtada, Rida;Herce, Henry D;Yin, Daniel J;Moroco, Jamie A;Wales, Thomas E;Engen, John R;Walensky, Loren D
通讯作者：
Walensky, Loren D