Preclinical Analyses of NAD Kinase as a Redox Vulnerability for the Treatment of Pancreatic Cancer

NAD 激酶作为氧化还原弱点治疗胰腺癌的临床前分析

• 批准号: 10062483
• 负责人: Sarah H. ELSEA
• 金额: $ 45.27万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-05 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10062483
• 关键词: Acute; Address; Adjuvant; Alleles; Antioxidants; Apoptosis; Attenuated; Automobile Driving; Bar Codes; Biological Assay; Cell Death; Cell Line; Cell model; Cells; Cellular Stress; Chemoresistance; Clinical; Collection; Colorectal Cancer; Cytoplasm; Data; Development; Diagnostic; Disease; Electroporation; Epigenetic Process; Future; Gene Frequency; Genes; Genetic; Genetic Screening; Genetically Engineered Mouse; Glutamine; Glutathione; Growth; KRAS2 gene; KRASG12D; Maintenance; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Metabolic; Metabolic Pathway; Modeling; Molecular; Mus; Mutate; Mutation; NAD+ kinase; NADP; Normal Cell; Oncogenes; Organism; Oxidants; Oxidation-Reduction; Oxidative Stress; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Phenotype; Phosphotransferases; Positioning Attribute; Production; Protein Kinase; Reactive Oxygen Species; Reagent; Resistance; Role; Stress; Technology; Testing; Therapeutic; Therapeutic Index; Tissue Microarray; Tissues; Toxic effect; United States; Work; Xenograft procedure; aging population; anticancer research; base; cancer cell; cell growth; chemotherapeutic agent; chemotherapy; colon cancer patients; conditional knockout; cost effective; gemcitabine; improved outcome; in vivo; insight; kinase inhibitor; malignant breast neoplasm; member; mortality; mouse model; neoplastic cell; novel; overexpression; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pre-clinical; response; screening; therapeutic target; tumor; tumor growth; tumor progression

Project Summary A major effort in cancer research is focused on identifying genes directly responsible for promoting cancer progression (referred to here as “drivers”). Perhaps nowhere is this need more acute than for pancreatic ductal adenocarcinoma (PDAC), a notoriously aggressive disease without durable treatment options. Identifying PDAC drivers and understanding their mechanism-of-action is critically important, as this information could inform new PDAC diagnostics and therapeutics. To identify such PDAC drivers, we developed novel genetic screening technologies to identify genes found mutated in PDAC that functionally cooperate with KRAS, the major driver gene found in pancreatic cancer, to promote PDAC tumor development in mice. Our approach identified the NAD Kinase (NADK), which is known in other organisms to influence redox metabolic pathways that regulate cell growth and resistance to growth-related oxidative stress. Our preliminary results indicate that NADK activation robustly drives PDAC initiation and growth, and NADK depletion significantly decreases PDAC growth concomitant with high oxidative stress owning to changes in redox state. Recent work by others has demonstrated the importance of redox pathways such as the glutamine reprogramming pathway (GRP) in promoting and maintaining PDAC growth. We hypothesize that interplay between the GRP and NADK activity centrally influences redox state and PDAC growth. We further hypothesize that NADK represents a redox vulnerability, as inhibiting NADK in patient tumors would serve as a means to selectively kill PDAC cells or sensitize them to cancer chemotherapeutics. In Aim1 we will use a large panel of PDAC cell lines and tumors genetically modified to inducibly express or deplete NADK and GRP expression to evaluate their relative roles in influencing redox state, oxidative stress and PDAC growth in culture assays and mice. In addition, we will examine the combined effect of NADK depletion and gemcitabine, a first-line PDAC chemotherapy agent and inducer of oxidative stress, to determine whether adjuvant use of NADK inhibitors would synergize with gemcitabine to kill PDAC cells. Finally, we will measure relative levels of NADK protein and oxidative stress on clinically- and genomically-annotated PDAC patient tumors, work intended to correlate these markers with gemcitabine response to provide insight on NADK inhibitor responder (patient) identification. In Aim 2 we will evaluate the in vivo role of NADK and therapeutic potential of NADK inhibitors by employing a novel electroporation model that allows rapid and cost-effective NADK expression and depletion in the context of activated KRAS in the mouse pancreas. In addition, we will examine NADK’s role in PDAC development and maintenance of tumor redox state using a genetically engineered mouse model of PDAC harboring a NADK conditional knockout allele. In the future, these models will also serve as a platform for testing additional chemotherapeutic agents (e.g., gemcitabine) in the context of targeted NADK depletion.

项目摘要 癌症研究的主要努力集中在识别直接促进癌症的基因 进展（这里称为“驱动程序”）。也许没有比胰管导管更急切的地方 腺癌（PDAC），一种臭名昭著的侵略性疾病，没有持久的治疗选择。识别 PDAC驱动程序和理解其行动机制至关重要，因为此信息可能 告知新的PDAC诊断和治疗。为了识别这种PDAC驱动因素，我们开发了新的遗传 筛选技术以鉴定在PDAC中发现的基因与KRAS合作的PDAC突变， 在胰腺癌中发现的主要驱动基因，可促进小鼠PDAC肿瘤的发展。我们的方法 确定了NAD激酶（NADK），在其他生物体中已知以影响氧化还原代谢途径 调节细胞生长和对生长相关氧化物应激的抗性。我们的初步结果表明 NADK激活稳健地驱动PDAC倡议和增长，而NADK耗竭显着下降 PDAC的生长与氧化应激具有高的氧化还原状态变化。别人最近的工作 已经证明了氧化还原途径的重要性，例如谷氨酰胺重编程途径（GRP） 促进和维持PDAC增长。我们假设GRP和NADK活动之间的相互作用 集中影响氧化还原状态和PDAC增长。我们进一步假设NADK代表氧化还原 脆弱性，因为在患者肿瘤中抑制NADK将作为选择性杀死PDAC细胞或 使它们对癌症化学治疗剂敏感。在AIM1中，我们将使用大型PDAC细胞系和肿瘤面板 通常修改以诱导表达或复制NADK和GRP表达以评估其相对角色 影响氧化还原状态，氧化应激和PDAC在培养测定和小鼠中的生长。此外，我们将 检查NADK部署和吉西他滨的综合作用，吉西他滨，一线PDAC化学疗法和 氧化应激的诱导剂，以确定调整NADK抑制剂的使用是否会与 吉西他滨杀死PDAC细胞。最后，我们将测量NADK蛋白的相对水平和氧化应激上的相对水平。 临床和基因宣布的PDAC患者肿瘤，旨在将这些标记与这些标记相关的工作 吉西他滨反应可提供对NADK抑制剂响应者（患者）识别的见解。在目标2中，我们将 通过采用新颖 电穿孔模型允许在 在小鼠胰腺中激活KRAS。此外，我们将研究NADK在PDAC开发中的作用， 使用携带NADK的PDAC的一般工程鼠标模型维护肿瘤氧化还原状态 有条件的淘汰等位基因。将来，这些模型还将作为测试其他模型的平台 在靶向NADK耗竭的情况下，化学治疗剂（例如，吉西他滨）。