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Investigation of NRF2-Dependent Metabolic Liabilities

NRF2 依赖性代谢负担的研究

基本信息

批准号：
10427369
负责人：
Gina Marie DeNicola
金额：
$ 39.35万
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：
2018
资助国家：
美国
起止时间：
2018-07-03 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10427369
关键词：
Agar Amino Acids Antioxidants 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 Lead 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 Oxides Pathway interactions Patients Positioning Attribute Process Production Proteins 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 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，并为癌症治疗带来希望。然而，控制这些代谢改变的潜在机制及其对肿瘤维持的贡献普遍缺乏。 NRF2 及其负调节因子 KEAP1 的突变存在于 15-34% 的非小细胞中。细胞肺癌 (NSCLC) 并导致组成型 NRF2 活性。许多 NRF2 监管的过程集中于由氨基酸合成的抗氧化剂谷胱甘肽的生产和利用半胱氨酸、甘氨酸和谷氨酸。在我们之前的工作中，我们发现谷胱甘肽的合成对于 NRF2 在肿瘤发生过程中的促增殖作用。此外，我们发现NRF2促进新陈代谢重新布线以增加甘氨酸的可用性以支持谷胱甘肽的合成。定义 NRF2 的组成方式活性诱导代谢重编程以支持 GSH 合成，我们通过基因工程产生，癌症突变 NRF2D29H 和 KEAP1R554Q 的条件敲入小鼠模型。我们的初步 NRF2 调节的代谢分析表明半胱氨酸双加氧酶 (CDO1) 限制谷胱甘肽 NRF2 产生，产生有毒副产物，并在肿瘤发生过程中被沉默。我们已经建立了一个用于分析含硫代谢物的代谢组学平台和基因工程 NRF2 和 KEAP1 突变肺癌小鼠模型用于体内研究，现在准备定义肿瘤 CDO1 及其代谢物在肺肿瘤发生过程中的抑制作用。在目标 1 中，我们将通过以下方式检查 CDO1 是否拮抗 NRF2 调节的抗氧化反应：耗尽半胱氨酸。在目标 2 中，我们将检查 CDO1 表达对具有 NRF2 活性的细胞的选择性毒性，因为毒性副产品生产。在目标 3 中，我们将使用我们的基因工程研究 CDO1 丢失是否会促进肺部肿瘤发生 KEAP1 和 NRF2 突变小鼠肺肿瘤模型和患者肿瘤样本。该项目的最终目标和总体影响是将 CDO1 描述为一种新型代谢负担针对具有 NRF2/KEAP1 突变的肿瘤，以确定患者治疗干预的机会这些突变目前缺乏靶向治疗。