Differential role of different NAD+ kinase Isoforms in melanoma metastasis

不同 NAD 激酶亚型在黑色素瘤转移中的不同作用

• 批准号: 10436014
• 负责人: Elena Piskounova
• 金额: $ 38.77万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10436014
• 关键词: Adopted; Affinity Chromatography; Anoikis; Antioxidants; Automobile Driving; Binding; Blood Circulation; Cancer Etiology; Cancer Patient; Cancer cell line; Carbon; Cell Line; Cells; Cessation of life; ChIP-seq; Clustered Regularly Interspaced Short Palindromic Repeats; Cytoplasm; Data; Dependence; Disease; Disease Outcome; Environment; Enzymes; Event; Genetic; Homeostasis; Human; Immunocompromised Host; In Situ; Malignant Neoplasms; Malignant neoplasm of pancreas; Melanoma Cell; Metabolic; Metabolic Pathway; Metastatic to; Mitochondria; Modeling; Molecular; Mus; Mutation; NAD+ kinase; NADP; Nature; Neoplasm Circulating Cells; Neoplasm Metastasis; Nodule; Normal Cell; Normal tissue morphology; Organ; Organelles; Oxidation-Reduction; Oxidative Stress; Pathway interactions; Patient-Focused Outcomes; Patients; Physiological; Primary Neoplasm; Production; Protein Isoforms; Protein Sequence Analysis; Reactive Oxygen Species; Reduced Glutathione; Regulation; Regulatory Element; Reporter; Resistance; Role; Signal Transduction; Stress; Testing; Therapeutic Intervention; Transcription Initiation Site; Transcriptional Regulation; Up-Regulation; Visceral; Work; cancer cell; cancer survival; cell regeneration; cell type; clinical predictors; clinically relevant; effective therapy; experience; in vitro Model; in vivo; melanoma; metabolic profile; migration; mortality; mouse model; neoplastic cell; new therapeutic target; novel; overexpression; patient derived xenograft model; promoter; response; sensor; subcutaneous; targeted treatment; transcription factor; tumor

PROJECT SUMMARY Metastasis is responsible for more than 90% of cancer patient mortality yet there are no therapies that specifically target metastatic disease. Many of the current in vitro models of metastasis focus on the molecular mechanisms of migration, invasion and/or surviving anoikis, but cannot recapitulate the complexity of the environment in which metastasis occurs in vivo. Conversely, in mouse models of metastasis, it has been difficult to examine the molecular mechanisms that enable cells to proceed through each distinct step of metastasis due to limited material that can be isolated and infrequency of metastatic events in these models. For these reasons little is known about the challenges facing metastasizing cells in vivo, and how they are overcome. We have previously established a clinically relevant model of melanoma metastasis, using patient-derived xenografts (PDX) in immunocompromised mice, that recapitulates the outcome of the disease of the patient in mice, to dissect the metastatic cascade into distinct steps. Using this model, we have shown that metastasizing melanoma cells undergo reversible metabolic adaptations to withstand oxidative stress in part through an increased dependence on NADPH-generating enzymes in the one-carbon pathway. Our preliminary data also show an increase in NADP+ levels in metastatic nodules compared to subcutaneous tumors, suggesting an increase in de novo NADP+ synthesis. NADP+ is generated from NAD+ by NAD+ kinase (NADK). We observe higher levels of NADK in metastatic nodules compared to subcutaneous tumors, where metastatic nodules express the isoform of NADK with the highest activity, while subcutaneous tumors do not. We will test the hypothesis that metastasizing melanoma cells upregulate a specific isoform of NADK to increase NADP+ production, increase oxidative stress resistance and survival at different steps of the metastatic cascade. Using both melanoma cell lines and PDX tumor cells, Aim 1 will determine the role of different NADK isoforms in oxidative stress resistance, Aim 2 will define the mechanism of transcriptional regulation of NADK isoforms, and Aim 3 will establish the role of different NADK isoforms as metastatic drivers in vivo. In addition, Aim 3 will test how perturbation of oxidative stress in different organelles impacts metastasis. Together this work will significantly contribute to our understanding of a novel mechanism of metabolic plasticity through upregulation of a specific NADK isoform and identify organelle-specific metabolic pathways as novel therapeutically targetable vulnerabilities in melanoma metastasis.

项目摘要 转移负责超过90％的癌症患者死亡率，但没有具体的疗法 靶向转移性疾病。当前许多转移的体外模型都集中在分子机制上 移民，入侵和/或幸存的厌食症，但不能概括环境的复杂性 转移发生在体内。相反，在转移的鼠标模型中，很难检查 分子机制，使细胞能够通过有限的每个不同的转移步骤进行 在这些模型中可以隔离的材料和转移性事件的不频率。由于这些原因，几乎没有 知道体内转移细胞面临的挑战以及如何克服。我们以前有 使用患者衍生的异种移植物（PDX）在临床上相关的黑色素瘤转移模型 免疫功能低下的小鼠，概括了小鼠疾病的结果，以剖析 转移级联分为不同的步骤。使用此模型，我们已经表明转移黑色素瘤细胞 通过增加的依赖性，经过可逆的代谢适应以承受氧化应激 在单碳途径中的NADPH生成酶上。我们的初步数据也显示 与皮下肿瘤相比，转移性结节中的NADP+水平，表明从头增加 NADP+合成。 NADP+由NAD+激酶（NADK）从NAD+生成。我们观察到较高的纳德克 在转移性结节与皮下肿瘤相比，转移性结节表达同工型 活性最高的NADK，而皮下肿瘤没有。我们将测试转移的假设 黑色素瘤细胞上调NADK的特定亚型增加NADP+产生，增加氧化应激 在转移性级联反应的不同步骤上的抗性和生存。 使用黑色素瘤细胞系和PDX肿瘤细胞，AIM 1将确定不同NADK同工型的作用 在氧化应激抗性中，AIM 2将定义NADK同工型转录调控的机制， AIM 3将确定不同NADK同工型作为体内转移驱动因素的作用。此外，AIM 3将 测试不同细胞器中氧化应激的扰动如何影响转移。这项工作将 通过上调有助于我们理解我们对代谢可塑性的新机制 特定的NADK同工型，并确定细胞器特异性代谢途径是新颖的治疗目标 黑色素瘤转移的脆弱性。