Mapping druggable co-dependency pathways in NRF2-driven lung cancers

绘制 NRF2 驱动的肺癌的药物共依赖性途径

基本信息

批准号：
10115633
负责人：
Liron Bar-Peled
金额：
$ 24.9万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-01 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10115633
关键词：
Address Advanced Malignant Neoplasm Anchorage-Independent Growth Antioxidants Applications Grants Binding Biochemical Biochemical Pathway Biological Biology Cancer Cell Growth Cancer Model Cancer Patient Cancer cell line Cell Proliferation Cells Chemicals Clustered Regularly Interspaced Short Palindromic Repeats Complex Cysteine Dependence Disease Doxycycline Drug Metabolic Detoxication Drug Targeting Environment Equilibrium Family Foundations Gene Expression Genes Genetic Genetic Transcription Goals Growth Lead Left Libraries Lung Neoplasms Malignant Neoplasms Malignant neoplasm of lung Maps Mediating Metabolic Metabolic Pathway Methods Mus Mutation NR0B1 gene Non-Small-Cell Lung Carcinoma Nuclear Orphan Receptor Oncogenic Orphan Output Oxidation-Reduction Pathway interactions Patients Pharmacology Proliferating Protein Binding Domain Proteins Proteomics Reactive Oxygen Species Research Role Signal Pathway Signal Transduction Stress Technology Therapeutic Transcriptional Activation Up-Regulation Xenograft Model Zinc Fingers anti-cancer therapeutic beta catenin bropirimine cancer cell cell growth chemoproteomics druggable target in vivo inhibitor/antagonist knock-down lung cancer cell lung xenograft next generation programs protein function protein protein interaction receptor response small molecule small molecule inhibitor targeted agent tool transcription factor tumor tumor xenograft tumorigenesis

项目摘要

Project Summary Rapidly proliferating cancer cells generate reactive oxygen species (ROS) that if left unchecked inhibit cell growth. To counter this stress, cancer cells and in particular non small cell lung cancers (NSCLC) rely on the activation of the NRF2 transcription factor, leading to the massive upregulation of key metabolic and detoxification proteins needed to restore redox balance. While directly targeting NRF2 with chemical inhibitors is challenging, we hypothesized that hyperactivation of this pathway would lead to an alteration of specific signaling and metabolic pathways required for the proliferation of these cells (co-dependencies), which themselves could be inhibited with small molecules. To identify these co-dependencies in NSCLC, we enriched for proteins containing reactive cysteines, which can be used as a chemical handle to develop inhibitors. This chemical proteomics screen identified hundreds of reactive cysteines regulated by NRF2, including a cryptic cysteine (C274) in the orphan receptor NR0B1. NR0B1 expression is severely restricted to those NCSLC cells and patient tumors with deregulated NRF2 signaling, where it functions as part of multimeric transcriptional complex to support the NRF2 gene expression program. As C274 in NR0B1 is necessary for NR0B1-complex formation, we exploited this residue to develop a small molecule inhibitor that covalently binds to it, subsequently disrupting the protein-protein interactions of NR0B1 and blocking the growth of NRF2-dependent cells, but not NRF2-independent cells. Thus, we have revealed NR0B1 as a druggable co-dependency of the NRF2 pathway. In this grant application, we build on our research on NR0B1 and further identify co-dependent pathways with NRF2 that can be pharmacologically interrogated. Using a powerful chemoproteomic framework, we will comprehensively define NRF2 co-dependencies by: 1) mapping the landscape of cysteine reactivity regulated by NRF2 in lung xenograft models, allowing us to identify cysteines on key proteins in the NRF2 pathway, which may become targetable opportunities in vivo 2) undertaking a small molecule screen to identify compounds that selectively inhibit the proliferation of NRF2-dependent NSCLC cell lines. Importantly, integrating a chemoproteomic platform into this screen, will allow for the rapid identification of co-dependent proteins, offering an unparalleled map of druggable NRF2 co-dependencies. The research proposed herein, takes full advantage of advanced cancer models and chemoproteomic technologies to reveal pharmacologically tractable proteins which are needed for the proliferation of NRF2-addicted cells and may provide a generalizable platform for inhibiting genetically defined cancers. These studies will not only provide a comprehensive understanding of NRF2 biology but might also lay the foundation for translational therapeutics benefiting lung cancer patients with deregulated NRF2 signaling.

项目概要快速增殖的癌细胞会产生活性氧 (ROS)，如果不加以控制，会抑制细胞生长为了对抗这种压力，癌细胞，特别是非小细胞肺癌（NSCLC）依赖于 NRF2 转录因子的激活，导致关键代谢和功能的大量上调恢复氧化还原平衡所需的解毒蛋白，同时用化学抑制剂直接靶向 NRF2。具有挑战性，我们假设该途径的过度激活会导致特定的改变这些细胞增殖所需的信号传导和代谢途径（相互依赖性），它们本身可能会受到小分子的抑制。为了确定 NSCLC 中的这些相互依赖性，我们丰富了用于含有反应性半胱氨酸的蛋白质，可用作开发抑制剂的化学处理剂。化学蛋白质组学筛选鉴定出数百个受 NRF2 调节的反应性半胱氨酸，包括一个神秘的孤儿受体 NR0B1 中的半胱氨酸 (C274)。NR0B1 的表达严格限于那些 NCSLC 细胞。以及 NRF2 信号传导失调的患者肿瘤，它作为多聚转录的一部分 NR0B1 复合物支持 NRF2 基因表达程序，因为 NR0B1 中的 C274 对于 NR0B1 复合物是必需的。形成，我们利用这个残基开发了一种与其共价结合的小分子抑制剂，随后破坏 NR0B1 的蛋白质-蛋白质相互作用并阻断 NRF2 依赖性细胞的生长细胞，但不是 NRF2 独立细胞。因此，我们发现 NR0B1 是一种可药物依赖性的细胞在本次拨款申请中，我们以 NR0B1 的研究为基础，进一步确定了相互依赖性。使用强大的化学蛋白质组学可以对 NRF2 的通路进行药理学研究。框架中，我们将通过以下方式全面定义 NRF2 相互依赖性：1）绘制半胱氨酸的图谱在肺异种移植模型中，反应性受 NRF2 调节，使我们能够识别肺异种移植模型中关键蛋白质上的半胱氨酸。 NRF2 通路，可能成为体内的目标机会 2) 进行小分子筛选重要的是，鉴定出选择性抑制 NRF2 依赖性 NSCLC 细胞系增殖的化合物。将化学蛋白质组学平台集成到该屏幕中，将允许快速识别相互依赖的蛋白质，提供了无与伦比的可药物 NRF2 相互依赖性图谱。本文提出的研究，充分利用先进的癌症模型和化学蛋白质组学技术来揭示 NRF2成瘾细胞增殖所需的药理学易处理的蛋白质，并且可能这些研究不仅提供了抑制基因癌症的通用平台。对 NRF2 生物学的全面了解，也可能为转化治疗奠定基础 NRF2 信号传导失调使肺癌患者受益。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Pharmacological convergence reveals a lipid pathway that regulates C. elegans lifespan.

药理学趋同揭示了调节线虫寿命的脂质途径。

DOI：
发表时间：
2019
期刊：
影响因子：
14.8
作者：
Chen, Alice L;Lum, Kenneth M;Lara;Ogasawara, Daisuke;Cognetta 3rd, Armand B;To, Alan;Parsons, William H;Simon, Gabriel M;Desai, Arshad;Petrascheck, Michael;Bar;Cravatt, Benjamin F
通讯作者：
Cravatt, Benjamin F

Chemical Proteomics Identifies Druggable Vulnerabilities in a Genetically Defined Cancer.

化学蛋白质组学识别基因定义的癌症中的药物脆弱性。

DOI：
发表时间：
2017-10-19
期刊：
影响因子：
64.5
作者：
Bar;Kemper, Esther K;Suciu, Radu M;Vinogradova, Ekaterina V;Backus, Keriann M;Horning, Benjamin D;Paul, Thomas A;Ichu, Taka;Svensson, Robert U;Olucha, Jose;Chang, Max W;Kok, Bernard P;Zhu, Zhou;Ihle, Nathan T;Dix, Melissa M
通讯作者：
Dix, Melissa M