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

基本信息

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
项目类别：
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&apos -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

项目摘要

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，导致在美国，每年有超过 48,000 例 PDAC 肿瘤因多种遗传和生理原因而死亡。促进剧毒活性氧 (ROS) 产生的特性，这些活性氧可以相互作用因此，这些肿瘤严重依赖 ROS 解毒程序。它利用硫醇基团来减轻定义 ROS 的电子不平衡。源自半胱氨酸，半必需氨基酸，具有含硫醇的侧链，最终是半胱氨酸。外源性半胱氨酸来源于膳食，导入是ROS解毒的关键步骤在本次资助的第一期中，我们测试了 PDAC 依赖于半胱氨酸导入的假设。我们发现半胱氨酸的消耗，可以通过介质的消耗或半胱氨酸的抑制来实现。摄取，导致铁死亡的诱导，这是一种由于氧化损伤而发生的调节性坏死我们证明半胱氨酸耗尽可以选择性诱导 PDAC 铁死亡。使用遗传和药理学方法对体内肿瘤进行研究，我们扩展了机制通过证明辅酶 A（半胱氨酸衍生物）的消耗是必要的来了解铁死亡用于诱导胰腺肿瘤细胞的铁死亡。在这里，我们将扩展这些发现，以增强 PDAC 中半胱氨酸消耗的影响。将研究 xC– 系统抑制与外源胱氨酸降解之间的机制差异然后我们将直接比较这些酶的临床前功效。 PDAC 基因工程模型中的方法，利用一种新的、高效的系统 xC– 抑制剂。其次，我们将通过研究进一步研究辅酶 A 在调节铁死亡中的作用。肿瘤微环境中高水平的辅酶 A 是否有助于抑制恶性肿瘤中的铁死亡这导致了针对 ENPP 蛋白的策略，这是（间接）导入所必需的。第三，我们将确定铁蛋白中游离铁的释放是否通过自噬可以在半胱氨酸耗尽的情况下促进铁死亡，这导致了激活策略。通过 MEK 抑制自噬增加脂质 ROS 的产生并对铁死亡敏感对于这些主题中的每一个，我们都将研究基础科学机制和转化。这些研究将利用 PDAC 基因工程小鼠模型以及新型肿瘤。我们小组开发的外植体模型，其中人或鼠 PDAC 样本在一个完整的培养皿中培养。这些研究将共同进一步阐明反应机制。胰腺导管腺癌的半胱氨酸消耗策略。