Metabolic basis of the NADPH-independent disulfide reductase system in mouse liver

小鼠肝脏中不依赖 NADPH 的二硫键还原酶系统的代谢基础

基本信息

批准号：
10056616
负责人：
Gina Marie DeNicola
金额：
$ 44.73万
依托单位：
MONTANA STATE UNIVERSITY - BOZEMAN
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-14 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10056616
关键词：
Active Sites Adenosine Affect Anabolism Biliary Biology Bypass Carbon Catabolism Cells Coenzyme A Consumption Creatine Cysteine DNA DNA Methylation Dependence Disease Disulfides Enzymes Excretory function Feedback Folic Acid Future Generations Glutamates Glutathione Glutathione Disulfide Glutathione Reductase Glycine Health Hepatic Hepatocyte Homeostasis Inflammatory Institution Investigation Knockout Mice Knowledge Label Lead Lecithin Lipids Liver Malignant Neoplasms Mammalian Cell Metabolic Metabolic Pathway Metabolism Methionine Methylation Methyltransferase Microbe Modeling Mus NADP Natural regeneration Nerve Degeneration Normal Cell Nutrient Organoids Outcome Oxidants Oxidation-Reduction Oxidative Stress Oxidoreductase Pathogenicity Pathway interactions Patients Peripheral Physiological Positioning Attribute Predisposition Process Production Proteins RNA Reaction Regulation Resources Role Route S-Adenosylhomocysteine Serine Source Stable Isotope Labeling Stress Sulfur Sulfur Amino Acids Sulfur Metabolism Pathway System Testing Therapeutic Tissues Toxin Translations Vision Work amino acid metabolism base carbon skeleton clinical application combat histone methylation liver metabolism metabolomics methyl group mouse model novel therapeutic intervention oxidation oxidative damage pathogen prevent reaction rate regenerative response stable isotope therapeutic target thioredoxin reductase thioredoxin reductase 1 uptake

Active Sites Adenosine Affect Anabolism Biliary Biology Bypass Carbon Catabolism Cells Coenzyme A Consumption Creatine Cysteine DNA DNA Methylation Dependence Disease Disulfides Enzymes Excretory function Feedback Folic Acid Future Generations Glutamates Glutathione Glutathione Disulfide Glutathione Reductase Glycine Health Hepatic Hepatocyte Homeostasis Inflammatory Institution Investigation Knockout Mice Knowledge Label Lead Lecithin Lipids Liver Malignant Neoplasms Mammalian Cell Metabolic Metabolic Pathway Metabolism Methionine Methylation Methyltransferase Microbe Modeling Mus NADP Natural regeneration Nerve Degeneration Normal Cell Nutrient Organoids Outcome Oxidants Oxidation-Reduction Oxidative Stress Oxidoreductase Pathogenicity Pathway interactions Patients Peripheral Physiological Positioning Attribute Predisposition Process Production Proteins RNA Reaction Regulation Resources Role Route S-Adenosylhomocysteine Serine Source Stable Isotope Labeling Stress Sulfur Sulfur Amino Acids Sulfur Metabolism Pathway System Testing Therapeutic Tissues Toxin Translations Vision Work amino acid metabolism base carbon skeleton clinical application combat histone methylation liver metabolism metabolomics methyl group mouse model novel therapeutic intervention oxidation oxidative damage pathogen prevent reaction rate regenerative response stable isotope therapeutic target thioredoxin reductase thioredoxin reductase 1 uptake

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项目摘要

What is known: Disulfide reduction-fueled enzymes s!upport homeostasis and combat oxidative damage that contributes to neurodegeneration, inflammatory diseases, and cancer. NADPH provides the reducing power for most anabolic and cytoprotective reduction reactions, yet only two enzymes can use NADPH to reduce cytosolic disulfides: thioredoxin reductase-1 (TrxR1) and glutathione reductase (Gsr) 1. Both TrxR1 and Gsr have active sites that are dominantly inhibited by electrophilic toxins and oxidants 2, 3. In Co-PI Schmidt’s lab, mice with TrxR1/Gsr-null livers uncovered unexpected robustness in the disulfide reductase systems, including an NADPH-independent pathway that uses catabolism of methionine (Met) to sustain redox homeostasis 4. Importantly, this pathway is also thought to sustain normal cells under oxidative or electrophilic stress 5. Met and Cys are the 2 sulfur (S)-amino acids found in proteins, but S-containing molecules synthesized from Met or Cys, including S-adenosyl-Met (SAM), glutathione (GSH), CoA, and others, are also important in redox, detox, energetics, biosynthesis, regulation, and other processes. Co-PI DeNicola has been studying the roles of altered S-amino acid metabolism in sustaining some cancers6. These studies are revealing how some cancers use altered S-amino acid redox metabolism, which could uncover targetable cancer-specific susceptibilities. Unresolved questions: It remains unknown how other metabolic activities, including those that directly utilize Met or Cys, as well as more peripheral systems that either (i) supply resources to these pathways; (ii) depend upon these pathways; or (iii) might, in some conditions, compete with these pathways for substrates, are realigned to help cells survive stress. We hypothesize that conversion to Met-dependence involves realignment of diverse metabolic pathways. This work is significant because a better understanding of these processes will uncover processes that can be therapeutically targeted to either specifically increase the robustness of critical cells under oxidative or toxic stress, or specifically increase the vulnerability of pathogenic cells in cancer or inflammatory diseases. New preliminary investigations in this resubmission demonstrate our ability to perform stable isotope flux labeling studies in whole mice and in mouse-derived hepatic organoids. What is proposed: In this revised multi-institution collaborative project, we will define the metabolic pathway realignments that occur when hepatocytes switch from NADPH-dependent to -independent disulfide reduction. We propose 3 Specific Aims: Aim 1, Define how NADPH- versus Met-fueled disulfide reductase homeostasis influences S-metabolism prioritization. Aim 2, Define how re-wiring of serine metabolism supports Met-fueled disulfide reductase homeostasis. Aim 3, Test whether Met-dependent survival increases the activity and dependence on liver methyltransferases. Anticipated outcomes, value: This project will help us understand how global shifts in hepatic metabolism occurs in response to severe oxidative or electrophilic stress in liver, and how this helps sustain health.

已知的：二硫化物还原燃料的酶S！Upport稳态和战斗氧化损害有助于神经退行性，炎症性疾病和癌症。 NADPH提供了降低的能力大多数合成代谢和细胞保护性还原反应，但只有两个酶可以使用NADPH来减少胞质二硫化物：硫氧还蛋白还原酶-1（TRXR1）和谷胱甘肽还原酶（GSR）1。TRXR1和GSR均具有由亲电毒素和氧化剂2，3的活性位点在Co-Pi Schmidt的实验室中。随着TRXR1/GSR无效的生命，发现了二硫化物还原系统中的意外鲁棒性，包括一个 NADPH独立的途径，使用甲二氨酸（MET）的分解代谢来维持氧化还原稳态4。重要的是，该途径还被认为可以在氧化或亲电压力下维持正常细胞5。 Cys是在蛋白质中发现的2个硫（S） - 氨基酸，但是从Met或Cys合成的含S的分子，包括S-腺苷-MET（SAM），谷胱甘肽（GSH），COA等，在氧化还原，排毒，能量学，生物合成，调节和其他过程。 Co-Pi Denicola一直在研究改变的作用 S-氨基酸代谢在维持某些癌症6。这些研究揭示了一些癌症的使用方式 S-氨基酸氧化还原代谢的改变，可能会发现可靶向的癌症特异性敏感性。未解决的问题：尚不清楚其他代谢活动，包括直接利用的活动 MET或CYS，以及更多的外围系统（i）为这些途径提供资源；（ii）依赖在这些途径上；或（iii）在某些情况下，可能与这些基材的这些途径竞争，是重新调整以帮助细胞生存胁迫。我们假设转换向MET依赖性涉及重组各种代谢途径。这项工作很重要，因为对这些过程的更好理解将发现可以热靶向以特异性提高临界的鲁棒性的过程氧化物或毒性应激下的细胞，或特别增加了致病细胞在癌症或炎症性疾病。在此重新提交中的新初步调查表明了我们执行的能力稳定的同位素通量标记研究和小鼠衍生的肝脏类器官中的稳定的同位素标记研究。提出的内容：在经过修订的多机构合作项目中，我们将定义代谢途径当肝细胞从依赖NADPH依赖性的二硫化物还原切换时，发生重新调整。我们提出3个具体目的：AIM 1，定义NADPH与燃烧的二硫键还原酶稳态如何影响S-代谢优先级。 AIM 2，定义丝氨酸代谢的重新培养如何支持Met-Fuel 二硫化物减少了体内平衡。 AIM 3，测试元素依赖性生存是否增加了活性和依赖肝甲基转移酶。抗后果，价值：该项目将有助于我们了解全球肝素代谢的变化发生在肝脏中严重的氧化或亲电应激之内发生的，以及这如何帮助维持健康。