Investigation of NRF2-Dependent Metabolic Liabilities

NRF2 依赖性代谢负担的研究

基本信息

批准号：
10582332
负责人：
Gina Marie DeNicola
金额：
$ 37.15万
依托单位：
H. LEE MOFFITT CANCER CTR & RES INST
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10582332
关键词：
Agar Amino Acids Antioxidants Applications Grants Biology CRISPR/Cas technology Cancer Center Cancer Model Cancer Patient Cancer cell line Cells Cysteine Cysteine Metabolism Pathway Cysteine dioxygenase Cystine Data Engineering Enzymes Genetic Engineering Genetically Engineered Mouse Genets Glutamates Glutathione Glycine Goals Growth Human Investigation Knock-in Mouse Lung Adenocarcinoma Lung Capacity Lung Neoplasms Maintenance Malignant neoplasm of lung Mediating Metabolic Metabolic Control Metabolism Methylation Modeling Mutant Strains Mice Mutation Nature Non-Small-Cell Lung Carcinoma Outcome Oxidation-Reduction Oxidative Stress Pathway interactions Patients Positioning Attribute Process Production Proliferating Proteins Reaction Role Sampling Specimen Sulfites Sulfur System TXN gene Taurine Testing Therapeutic Intervention Toxic effect Work Xenograft Model cancer therapy cysteine sulfinate disulfide bond epigenetic silencing hypotaurine in vivo Model lung cancer cell lung tumorigenesis metabolomics mouse model mutant neoplastic cell novel parent grant protein expression response restoration targeted treatment transcription factor tumor tumor initiation tumorigenesis uptake

项目摘要

Metabolic reprogramming occurs during tumorigenesis and holds promise for cancer therapy. However, the underlying mechanisms that control these metabolic alterations and their contribution to tumor maintenance are generally lacking. Mutations in NRF2 and its negative regulator KEAP1 are found in 15-34% of non-small cell lung cancer (NSCLC) and lead to constitutive NRF2 activity. Many NRF2-regulated processes center on the production and utilization of the antioxidant glutathione, which is synthesized from the amino acids cysteine, glycine and glutamate. In our previous work, we found that glutathione synthesis was critical for the pro-proliferative effects of NRF2 during tumor initiation. Furthermore, we found that NRF2 promotes metabolic rewiring to increase glycine availability to support glutathione synthesis. To define how constitutive NRF2 activity induces metabolic reprogramming to support GSH synthesis, we generated genetically engineered, conditional knock-in mouse models of the cancer mutations NRF2D29H and KEAP1R554Q. Our preliminary analysis of NRF2-regulated metabolism indicates that cysteine dioxygenase (CDO1) limits glutathione production by NRF2, produces toxic byproducts, and is silenced during tumorigenesis. We have established a metabolomics platform for the analysis of sulfur-containing metabolites, and a genetically engineered NRF2 and KEAP1 mutant lung cancer mouse models for in vivo studies and are now poised to define the tumor suppressive role of CDO1 and its metabolites during lung tumorigenesis. In Aim 1 we will examine the whether CDO1 antagonizes the NRF2-regulated antioxidant response by depleting cysteine. In Aim 2 we will examine the selective toxicity of CDO1 expression to cells with NRF2 activity due to toxic byproduct production. In Aim 3 we will examine whether CDO1 loss promotes lung tumorigenesis using our genetically engineered KEAP1 and NRF2 mutant mouse lung tumor models and patient tumor samples. The ultimate goal and the overall impact of this project are to characterize CDO1 as a novel metabolic liability for tumors with NRF2/KEAP1 mutations in order to define opportunities for therapeutic intervention for patients with these mutations, which currently lack targeted therapy.

代谢重编程发生在肿瘤发生过程中，并有望接受癌症治疗。但是，控制这些代谢改变及其对肿瘤维持的贡献的基本机制通常缺乏。 NRF2中的突变及其负调节剂KEAP1在15-34％的非小型中发现细胞肺癌（NSCLC），并导致构型NRF2活性。许多NRF2调节的过程以抗氧化剂谷胱甘肽的产生和利用，该抗氧化剂是由氨基酸合成的半胱氨酸，甘氨酸和谷氨酸。在我们以前的工作中，我们发现谷胱甘肽合成对于 NRF2在肿瘤开始期间的促增强作用。此外，我们发现NRF2促进了代谢重新布线以增加甘氨酸的供应性，以支持谷胱甘肽合成。定义本构nrf2的方式活性诱导代谢重编程以支持GSH合成，我们生成了基因工程的，有条件的癌症突变小鼠模型NRF2D29H和KEAP1R554Q。我们的初步 NRF2调节的代谢的分析表明，半胱氨酸二加氧酶（CDO1）限制了谷胱甘肽 NRF2的生产产生有毒的副产品，并在肿瘤发生过程中被沉默。我们已经建立了用于分析含硫代谢物的代谢组学平台和基因工程的NRF2 和KEAP1突变肺癌小鼠模型用于体内研究，现在有望定义肿瘤 CDO1及其代谢产物在肺部肿瘤发生过程中的抑制作用。在AIM 1中，我们将检查CDO1是否通过耗尽半胱氨酸。在AIM 2中，我们将检查CDO1表达对具有NRF2活性细胞的选择性毒性副产品生产。在AIM 3中，我们将检查CDO1损失是否使用我们的基因工程来促进肺肿瘤发生 KEAP1和NRF2突变小鼠肺肿瘤模型和患者肿瘤样品。该项目的最终目标和整体影响是将CDO1描述为一种新型代谢责任对于具有NRF2/KEAP1突变的肿瘤，以定义患者治疗干预的机会这些突变目前缺乏针对性的治疗。