Targeting cysteine import to induce ferroptotic cell death in pancreatic cancer

靶向半胱氨酸输入诱导胰腺癌铁死亡细胞

• 批准号: 10446758
• 负责人: Kenneth P Olive
• 金额: $ 40.94万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-08 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10446758
• 关键词: 4' -phosphopantetheine; Acute; Alleles; Amino Acids; Antioxidants; Autophagocytosis; Basic Science; Biochemical Genetics; Cancer Etiology; Cell Death; Cell Line; Cells; Cellular Membrane; Cessation of life; Chemistry; Coenzyme A; Cyst; Cysteine; Cystine; Cytotoxic Chemotherapy; Data; Detoxification Process; Diet; Disulfides; Drug Metabolic Detoxication; Electrons; Engineering; Enzymes; FDA approved; Ferritin; Generations; Genetic; Genetic Engineering; Genetically Engineered Mouse; Glutamates; Goals; Grant; Human; Hypoxia; Immunotherapy; Intercellular Fluid; Interruption; Iron; KRAS2 gene; Knockout Mice; Lipids; MEK inhibition; MEKs; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of pancreas; Mammalian Cell; Mediating; Membrane; Membrane Lipids; Metabolism; Methods; Modeling; Molecular Biology; Mus; Mutant Strains Mice; Mutation; Necrosis; Normal Cell; Oncoproteins; Oxidation-Reduction; Oxidative Stress; Oxides; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Phenocopy; Phenotype; Physiological; Polyunsaturated Fatty Acids; Process; Production; Property; Proteins; Reaction; Reactive Oxygen Species; Regimen; Resistance; Role; Rupture; Sampling; Science; Side; Source; Stable Isotope Labeling; Sulfhydryl Compounds; System; Systems Biology; TP53 gene; Techniques; Testing; Tissues; Tumor Immunity; United States; Water; Work; anti-cancer; antiporter; clinical translation; comparative efficacy; detoxication; dietary; extracellular; improved; in vivo; inhibitor; macromolecule; mortality; mouse model; mutant; novel; oxidative damage; pancreatic cancer cells; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; peroxidation; preclinical efficacy; prevent; programs; recombinase; response; semi essential amino acid; targeted treatment; therapeutic target; therapeutically effective; translational approach; tumor; tumor metabolism; tumor microenvironment; uptake; 4' -phosphopantetheine; Acute; Alleles; Amino Acids; Antioxidants; Autophagocytosis; Basic Science; Biochemical Genetics; Cancer Etiology; Cell Death; Cell Line; Cells; Cellular Membrane; Cessation of life; Chemistry; Coenzyme A; Cyst; Cysteine; Cystine; Cytotoxic Chemotherapy; Data; Detoxification Process; Diet; Disulfides; Drug Metabolic Detoxication; Electrons; Engineering; Enzymes; FDA approved; Ferritin; Generations; Genetic; Genetic Engineering; Genetically Engineered Mouse; Glutamates; Goals; Grant; Human; Hypoxia; Immunotherapy; Intercellular Fluid; Interruption; Iron; KRAS2 gene; Knockout Mice; Lipids; MEK inhibition; MEKs; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of pancreas; Mammalian Cell; Mediating; Membrane; Membrane Lipids; Metabolism; Methods; Modeling; Molecular Biology; Mus; Mutant Strains Mice; Mutation; Necrosis; Normal Cell; Oncoproteins; Oxidation-Reduction; Oxidative Stress; Oxides; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Phenocopy; Phenotype; Physiological; Polyunsaturated Fatty Acids; Process; Production; Property; Proteins; Reaction; Reactive Oxygen Species; Regimen; Resistance; Role; Rupture; Sampling; Science; Side; Source; Stable Isotope Labeling; Sulfhydryl Compounds; System; Systems Biology; TP53 gene; Techniques; Testing; Tissues; Tumor Immunity; United States; Water; Work; anti-cancer; antiporter; clinical translation; comparative efficacy; detoxication; dietary; extracellular; improved; in vivo; inhibitor; macromolecule; mortality; mouse model; mutant; novel; oxidative damage; pancreatic cancer cells; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pancreatic neoplasm; peroxidation; preclinical efficacy; prevent; programs; recombinase; response; semi essential amino acid; targeted treatment; therapeutic target; therapeutically effective; translational approach; tumor; tumor metabolism; tumor microenvironment; uptake

Abstract: Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal human malignancies and is responsible for over 48,000 deaths per year in the United States. PDAC tumors have several genetic and physiological properties that promote the generation of highly toxic reactive oxygen species (ROS), which can interact destructively with cellular macromolecules. As a result, these tumors rely heavily on ROS detoxification programs which employ thiol groups to mitigate the electron imbalances that define ROS. Those thiols are ultimately derived from cysteine, a semi-essential amino acid with a thiol-containing side chain. As cysteine is ultimately sourced from dietary sources, the import of exogenous cysteine is a critical step in the ROS detoxification process. In the first term of this grant, we tested the hypothesis that PDAC depends on cysteine import for survival. We found that depletion of cysteine, either through depletion from the media or by inhibition of cysteine uptake, resulted in the induction of ferroptosis, a form of regulated necrosis that occurs due to oxidative damage to cellular membranes. We demonstrated that cysteine depletion could selectively induce ferroptosis in PDAC tumors in vivo using both genetic and pharmacological approaches and we expanded the mechanistic understanding of ferroptosis by demonstrating that depletion of coenzyme A (a cysteine derivative) is necessary for the induction of ferroptosis in pancreatic tumor cells. Here we will extend these findings in order to potentiate the effects of cysteine depletion in PDAC. First, we will study the mechanistic difference between inhibition of system xC– versus degradation of exogenous cystine with the engineered enzyme cyst(e)inase. We will then directly compare the preclinical efficacy of these approaches in a genetically engineered models of PDAC, making use of a new, highly potent system xC– inhibitor. Second, we will pursue our earlier findings on the role of coenzyme A in regulating ferroptosis by studying whether high levels of coenzyme A in the tumor microenvironment might help suppress ferroptosis in malignant epithelial cells. This leads to a strategy of targeting ENPP proteins, which are necessary for the (indirect) import of exogenous coenzyme A. Third, we will determine whether the release of free iron from ferritin through via autophagy can promote ferroptosis in the context of cysteine depletion. This leads to a strategy of activating autophagy via MEK inhibition to increase lipid ROS production and sensitize to ferroptosis in combination with cysteine depletion. For each of these topics, we will pursue both basic science mechanisms and translational strategies. The studies will make use of genetically engineered mouse models of PDAC as well as novel tumor explant models developed by our group in which human or murine PDAC samples are cultured intact in an engineered system for up to a week. Together, these studies will further elucidate the mechanisms of response to cysteine-depletion strategies in pancreatic ductal adenocarcinoma.

抽象的： 胰腺导管腺癌（PDAC）是最致命的人类恶性肿瘤之一，负责 在美国，每年超过48,000人死亡。 PDAC肿瘤具有几种遗传和生理 促进高毒活性氧（ROS）的产生的特性，可以相互作用 用细胞大分子破坏性。结果，这些肿瘤严重依赖ROS排毒程序 哪些员工硫醇组减轻了定义ROS的电子失衡。那些硫醇最终是 源自半胱氨酸，这是一种含有硫醇的侧链的半必需氨基酸。由于半胱氨酸最终 来自饮食来源，外源性半胱氨酸的进口是ROS排毒的关键步骤 过程。在这笔赠款的第一项中，我们检验了以下假设：PDAC取决于半胱氨酸进口 生存。我们发现半胱氨酸的消耗是通过培养基的耗竭或通过抑制半胱氨酸的耗竭 摄取，导致诱导铁铁病，这是一种受氧化损伤引起的调节坏死形式 到细胞膜。我们证明半胱氨酸的耗竭可以选择性地诱导PDAC的铁毒性。 使用遗传和药物方法的体内肿瘤，我们扩展了机械 通过证明辅酶A（半胱氨酸衍生物）的消耗是有必要的 为了诱导胰腺肿瘤细胞中的铁凋亡。 在这里，我们将扩展这些发现，以便潜在PDAC中半胱氨酸消耗的影响。首先，我们 将研究系统XC的抑制与外源性胱氨酸降解之间的机械差异 使用工程酶囊肿（E）inase。然后，我们将直接比较这些的临床前效率 使用新的，高潜在的系统XC抑制剂的普通工程模型中的方法。 其次，我们将通过研究辅酶A在调节铁蛋白方面的作用来追求我们的早期发现。 肿瘤微环境中高水平的辅酶A是否有助于抑制恶性肿瘤 上皮细胞。这导致了靶向ENPP蛋白的策略，这对于（间接）导入是必需的 外源辅酶A的A。第三，我们将确定从铁蛋白通过Via释放游离铁是否释放 自噬可以在半胱氨酸耗竭的背景下促进铁铁作用。这导致了激活的策略 通过MEK抑制自噬，以增加脂质ROS的产生和对铁凋亡的敏感性 半胱氨酸消耗。对于这些主题，我们将同时追求基础科学机制和翻译 策略。研究将利用PDAC的一般工程鼠标模型以及新型肿瘤 我们的小组开发的外植物模型，其中人类或鼠PDAC样品完好无损地培养 工程系统长达一周。总之，这些研究将进一步阐明反应的机制 在胰腺导管腺癌中的半胱氨酸排泄策略。