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

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.

摘要： 胰腺导管腺癌(PDAC)是人类最致命的恶性肿瘤之一，与 在美国，每年有超过4.8万人死亡。PDAC肿瘤有几种遗传学和生理学特征 促进产生剧毒的活性氧物种(ROS)的特性，这种ROS可以相互作用 对细胞大分子具有破坏性。因此，这些肿瘤严重依赖ROS解毒计划。 它们使用硫醇基团来缓解定义ROS的电子不平衡。这些硫醇最终是 来源于半胱氨酸，一种半必需氨基酸，含有硫醇侧链。因为半胱氨酸最终会 从饮食来源进口外源半胱氨酸是ROS解毒的关键一步 进程。在这笔赠款的第一个任期内，我们测试了PDAC依赖半胱氨酸进口的假设 生死存亡。我们发现半胱氨酸的耗尽，要么是通过从介质中耗尽，要么是通过抑制半胱氨酸 摄取，导致铁下垂的诱导，这是一种由于氧化损伤而发生的调节性坏死形式 到细胞膜。我们证明半胱氨酸的耗竭可以选择性地诱导PDAC的铁性下垂。 体内肿瘤的遗传和药理学方法，我们扩大了机制 通过证明辅酶A(一种半胱氨酸衍生物)的耗竭是理解铁性下垂的必要方法 用于诱导胰腺肿瘤细胞铁性下垂。 在这里，我们将扩展这些发现，以加强半胱氨酸耗竭对PDAC的影响。首先，我们 将研究抑制XC-系统和降解外源半胱氨酸之间的机理差异 用工程化酶包囊(E)酶。然后我们将直接比较这些药物的临床前疗效 在PDAC的基因工程模型中，利用一种新的、高度有效的系统XC-抑制剂的方法。 其次，我们将通过研究来继续我们早先关于辅酶A在调节铁性下垂中的作用的发现 肿瘤微环境中高水平的辅酶A是否有助于抑制恶性肿瘤患者的铁下垂 上皮细胞。这导致了以ENPP蛋白质为靶点的策略，这些蛋白质是(间接)进口所必需的 外源辅酶A。第三，我们将确定铁蛋白通过 在半胱氨酸耗竭的情况下，自噬可以促进铁性下垂。这导致了一种激活策略 通过抑制MEK自噬增加脂质ROS的产生并联合应用增敏铁性下垂 半胱氨酸耗尽。对于这些主题中的每一个，我们都将研究基础科学机制和翻译 战略。这些研究将利用基因工程小鼠的PDAC模型以及新的肿瘤 本课题组建立的外植体模型中，人或小鼠的PDAC样本在 长达一周的工程系统。总之，这些研究将进一步阐明反应的机制。 胰腺导管腺癌的半胱氨酸耗竭策略。