Nrf2 as a Master Regulator in Liver Disease Prevention and Therapy

Nrf2 作为肝病预防和治疗的主调节因子

• 批准号: 7788807
• 负责人: CURTIS DEAN KLAASSEN
• 金额: $ 37.51万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7788807
• 关键词: Active Sites; Animal Model; Anions; Anti-Inflammatory Agents; Anti-inflammatory; Antioxidants; Applications Grants; Attention; Basic Science; Biliary; Binding; Binding Sites; Cell Line; Cell Nucleus; Cells; Chemicals; Chimeric Proteins; Chinese People; Chromatin; Chromosomes, Human, Pair 10; Cirrhosis; Collaborations; Cysteine; Cytoprotection; Cytosol; Data; Development; Diet; Disease; Disease Outcome; Disease susceptibility; Drug Delivery Systems; Elements; Ensure; Epigenetic Process; Etiology; Excretory function; Exhibits; Fatty acid glycerol esters; Foundations; Functional disorder; Gene Expression; Gene Mutation; Gene Targeting; Genes; Genetic Models; Genetic Transcription; Genome; Glutathione; Goals; Green Fluorescent Proteins; Hepatic; Hepatocyte; Hepatotoxicity; Histone H3; Homologous Gene; Human; Image; Immune; Immunomodulators; Inflammation; Inflammatory; Inflammatory Response; Injury; Interferon Type II; Intervention; Knock-out; Knockout Mice; Lead; Lipid Peroxidation; Liver; Liver Failure; Liver Fibrosis; Liver diseases; Lysine; Malnutrition; Maps; Mediating; Medical; Messenger RNA; Metabolic Biotransformation; Modeling; Molecular; Monitor; Mus; Nuclear; Nuclear Translocation; Oleanolic Acid; Oxidative Stress; Pathogenesis; Pharmaceutical Preparations; Pharmacology and Toxicology; Phosphoric Monoester Hydrolases; Play; Poison; Preparation; Prevention; Prevention strategy; Prevention therapy; Proteins; Public Health; Regulation; Resolution; Role; Screening procedure; Signal Transduction; Solid; Specificity; Steatohepatitis; T-Lymphocyte; Techniques; Testing; Therapeutic; Therapeutic Intervention; Time; Toxic effect; Tumor Suppressor Genes; Ubiquitination; Up-Regulation; Wound Healing; adapter protein; analog; base; cell type; chromatin immunoprecipitation; cytokine; disorder prevention; drug development; experience; feeding; hepatotoxin; high throughput screening; innovation; liver pharmacology; mouse genome; mouse model; multidisciplinary; new therapeutic target; nonalcoholic steatohepatitis; novel; nuclear factor-erythroid 2; prevent; public health relevance; research study; response; small molecule libraries; tensin; therapeutic target; tool; toxicant; transcription factor

DESCRIPTION (provided by applicant): Current therapeutic interventions for treating liver diseases are very limited; identification of a therapeutic target is a prerequisite for developing new drugs to treat liver diseases. Oxidative stress and inflammation play key roles in the pathogenesis of liver diseases. Nuclear factor 2-related factor 2 (Nrf2), a transcription factor expressed in both hepatocytes and immune cells, is a key factor in anti-oxidative and anti-inflammatory responses. In quiescent hepatocytes, Nrf2 is sequestered in cytosol by kelch-like ECH-associated protein 1 (Keap1); in Keap1-knockdown mice, nuclear translocation of Nrf2 increases, resulting in activation of Nrf2 target genes. The importance of Nrf2 in cytoprotection is recognized. Nevertheless, the role of Nrf2 in liver pathophysiology has just recently gained attention. Our long-term goal is to identify a novel drug target and develop a therapeutic/preventive strategy for liver diseases. The objective of this R01 grant proposal is to investigate in-depth the role of Nrf2 in protecting against liver pathophysiology and understand its underlying molecular mechanisms. Based upon our strong preliminary data, our central hypothesis is that Nrf2 is a master regulator of liver protection through its multifaceted roles in biotransformation/disposition, antioxidation, tissue- repair, and anti-inflammation in multiple cell types in liver. Aim 1 will identify direct targets of Nrf2 regulation using ChIP-Sequencing and determine Nrf2-dependent mRNA induction in the Keap1-knockdown mouse. Aim 2 will test the hypothesis that Nrf2 activation protects against chemical, dietary, and genetic models of liver disease. We will use 3 genetic models of mice with differential Nrf2 activity, namely Nrf2-null, wild-type, and Nrf2-enhanced mice, to test liver disease susceptibility in 13 different models. Aim 3 will develop novel and specific Nrf2 activator lead candidates by high throughput screening in collaboration with the Office of Therapeutics, Discovery, and Development. Our proposed study is novel, because it utilizes 3 mouse lines with different Nrf2 activities to investigate the opposing effects of Nrf2 deficiency and Nrf2 activation on liver diseases induced via diverse mechanisms of toxicity. This proposal utilizes innovative techniques including ChIP-sequencing, multiplex mRNA analysis, and nuclear translocation high throughput screening in testing the 3 aims. Our proposed study is significant, because results from our experiments will provide solid evidence regarding the importance of Nrf2 deficiency and Nrf2 activation in liver pathophysiology and its underlying molecular mechanisms, setting the scientific foundation for developing Nrf2 activators as novel drugs to treat/prevent various liver diseases, such as chemical-induced liver injury, inflammatory liver diseases, steatohepatitis, and liver fibrosis/cirrhosis. PUBLIC HEALTH RELEVANCE: Liver disease is a common ailment, however in contrast to most other diseases, there are very few drugs for liver failure. We hypothesize that Nrf2, an essential transcription factor, is a drug target that can be used to prevent and treat a wide-range of liver ailments. This study will determine whether this drug target will be effective against a wide-spectrum of animal models of liver disease.

描述（由申请人提供）：目前治疗肝病的治疗干预措施非常有限；确定治疗靶点是开发治疗肝病新药的先决条件。氧化应激和炎症在肝脏疾病的发病机制中发挥着关键作用。核因子2相关因子2 (Nrf2) 是一种在肝细胞和免疫细胞中表达的转录因子，是抗氧化和抗炎反应的关键因子。在静止肝细胞中，Nrf2 被 kelch 样 ECH 相关蛋白 1 (Keap1) 隔离在细胞质中；在 Keap1 敲除小鼠中，Nrf2 的核易位增加，导致 Nrf2 靶基因激活。 Nrf2 在细胞保护中的重要性已得到认可。然而，Nrf2 在肝脏病理生理学中的作用最近才引起人们的关注。我们的长期目标是确定新的药物靶点并制定肝病的治疗/预防策略。这项 R01 拨款提案的目的是深入研究 Nrf2 在预防肝脏病理生理学方面的作用并了解其潜在的分子机制。基于我们强有力的初步数据，我们的中心假设是，Nrf2 通过其在肝脏多种细胞类型的生物转化/处置、抗氧化、组织修复和抗炎方面的多方面作用，成为肝脏保护的主要调节因子。目标 1 将使用 ChIP 测序确定 Nrf2 调节的直接靶标，并确定 Keap1 敲除小鼠中 Nrf2 依赖性 mRNA 的诱导。目标 2 将检验 Nrf2 激活可预防化学、饮食和遗传模型肝病的假设。我们将使用3种具有差异Nrf2活性的小鼠遗传模型，即Nrf2缺失小鼠、野生型小鼠和Nrf2增强小鼠，来测试13种不同模型的肝病易感性。 Aim 3 将与治疗、发现和开发办公室合作，通过高通量筛选开发新型、特异性 Nrf2 激活剂先导候选药物。我们提出的研究是新颖的，因为它利用具有不同 Nrf2 活性的 3 个小鼠系来研究 Nrf2 缺乏和 Nrf2 激活对通过不同毒性机制引起的肝脏疾病的相反影响。该提案利用 ChIP 测序、多重 mRNA 分析和核易位高通量筛选等创新技术来测试这 3 个目标。我们提出的研究意义重大，因为我们的实验结果将为Nrf2缺乏和Nrf2激活在肝脏病理生理学及其潜在分子机制中的重要性提供确凿的证据，为开发Nrf2激活剂作为治疗/预防各种肝病的新药奠定科学基础，例如化学性肝损伤、炎症性肝病、脂肪性肝炎和肝脏疾病。 纤维化/肝硬化。公共卫生相关性：肝病是一种常见疾病，但与大多数其他疾病相比，治疗肝衰竭的药物很少。我们假设 Nrf2（一种重要的转录因子）是可用于预防和治疗多种肝脏疾病的药物靶标。这项研究将确定该药物靶点是否对多种肝病动物模型有效。