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：
9891966
负责人：
Liron Bar-Peled
金额：
$ 22.88万
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/9891966
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 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.

项目摘要迅速增殖的癌细胞产生活性氧（ROS），如果不选中，则抑制细胞生长。为了应对这种压力，癌细胞，尤其是非小细胞肺癌（NSCLC）依靠 NRF2转录因子的激活，导致关键代谢和恢复氧化还原平衡所需的排毒蛋白。同时直接用化学抑制剂靶向NRF2 很具有挑战性，我们假设该途径的过度激活将导致特定的改变这些细胞增殖需要的信号传导和代谢途径（共依赖性），这是可以用小分子抑制自己。为了识别NSCLC中的这些共依赖性，我们丰富了对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手柄。这化学蛋白质组学筛查确定了数百种受NRF2调节的反应性囊肿，包括加密货币孤儿受体NR0B1中的半胱氨酸（C274）。 NR0B1表达严重限制在那些NCSLC细胞和患者肿瘤的NRF2信号传导，在该肿瘤中起作用，作为多项转录的一部分支持NRF2基因表达程序的复合物。由于NR0B1中的C274对于NR0B1复合是必需的形成，我们探索了这个住所，以开发一种与之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止NRF2依赖性的生长细胞，但不是NRF2独立的细胞。那我们已经揭示了NR0B1是可拖动的共同依赖性 NRF2途径。在此赠款应用程序中，我们以NR0B1的研究为基础，并进一步确定共同依赖性具有NRF2的途径，可以进行物理询问。使用功能强大的化学蛋白质组学框架，我们将通过以下方式全面定义NRF2共同依赖性：1）映射半胱氨酸的景观在肺部异种移植模型中由NRF2调节的反应性，使我们能够鉴定在关键蛋白上的半胱氨酸 NRF2途径，它可能成为体内的目标机会2）识别有选择地抑制NRF2依赖性NSCLC细胞系的增殖的化合物。重要的是，将化学蛋白质组学平台集成到此屏幕中，将允许快速识别共同依赖性蛋白质，提供无与伦比的NRF2共同依赖性图。本文提出的研究，充分利用高级癌症模型和化学蛋白质组学技术来解决在药理学上可触及的蛋白质，这是NRF2成瘾细胞增殖所需的，可能提供一个可推广的平台，用于抑制一般定义的癌症。这些研究不仅会提供对NRF2生物学的全面理解，但也可能为翻译治疗剂奠定基础受益于肺癌患者的NRF2信号传导。