Novel Redox-Associated Mechanisms Preventing Alcoholic Fatty Liver

预防酒精性脂肪肝的新型氧化还原相关机制

• 批准号: 9310228
• 负责人: Ying Chen
• 金额: $ 17.56万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-05 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9310228
• 关键词: 5' -AMP-activated protein kinase; Academic Training; Acetaldehyde; Acetic Acids; Advisory Committees; Alcohol dehydrogenase; Alcoholic Fatty Liver; Alcoholic Liver Diseases; Alcoholic liver damage; Alcohols; Anabolism; Animal Model; Antioxidants; Biochemical; Biological Markers; CYP2E1 gene; Catalytic Domain; Cause of Death; Cessation of life; Chronic; Cirrhosis; Clinical; Competence; Data; Development; Diagnostic; Disease; Drug Kinetics; Drug Metabolic Detoxication; Enzymes; Ethanol; Ethanol Metabolism; Ethanol toxicity; Event; Exhibits; Extrahepatic; Fatty Liver; Fibrosis; Free Radicals; GCLC gene; GCLM gene; Gene Expression; Gene Targeting; Generations; Genes; Glutamate-Cysteine Ligase; Glutathione; Goals; Hepatic; Hepatocyte; Immunologist; Individual; Inflammation; Inflammatory Response; Knock-out; Knockout Mice; Knowledge; Life Style; Link; Lipids; Liver; Liver diseases; Long-Term Effects; Mediating; Mentored Research Scientist Development Award; Mentors; Metabolic; Metabolic Pathway; Metabolic stress; Modeling; Molecular Biology; Mus; Nutritional; Oxidants; Oxidation-Reduction; Oxidative Stress; Oxidative Stress Induction; Pathogenesis; Pathogenicity; Pathway interactions; Phenotype; Play; Predisposition; Preventive; Protein Isoforms; Regulation; Research; Research Personnel; Research Project Grants; Resources; Role; STK11 gene; Scientist; Signal Pathway; Signal Transduction; Small Interfering RNA; Statistical Methods; Steatohepatitis; Stress; Sulfhydryl Compounds; Triglycerides; United States; Up-Regulation; Viral; Wild Type Mouse; alcohol abuse therapy; alcohol exposure; alcohol research; alcohol response; aldehyde dehydrogenases; base; biological adaptation to stress; career; economic impact; experimental study; feeding; health economics; knock-down; lipid metabolism; liver injury; member; mouse model; new therapeutic target; non-alcoholic fatty liver; novel; novel therapeutics; nuclear factor-erythroid 2; overexpression; prevent; problem drinker; programs; response; therapeutic target; tool; upstream kinase

PROJECT SUMMARY Cirrhosis-associated death is one of the leading causes of death in the United States and alcoholic fatty liver disease (AFLD) accounts for 48% of these deaths1. Despite its profound health and economic impact, the management of AFLD remains a challenging prospect because: (i) there are no effective diagnostic tools or biomarkers to assess individual susceptibility to AFLD development or progression to more severe clinical conditions, and (ii) no new therapeutic entities have been developed in the past four decades. The long term goal of this research project is to elucidate the redox-associated mechanisms involved in protecting the liver against AFLD and thereby identify potential novel preventive and/or therapeutic targets against this disease. Oxidative stress plays a central role in many pathways involved in the pathogenesis of AFLD. A major factor contributing to the development of oxidative stress is the depletion of glutathione (GSH), the most abundant non-protein thiol in the liver. Our preliminary studies using GCLM knockout (KO) mice demonstrate that ~85% deficiency in hepatic GSH renders mice protected from steatosis induced by chronic ethanol administration. These mice also exhibit: (i) enhanced capacity to metabolize ethanol and acetaldehyde, (ii) persistent oxidative stress and induction of nuclear factor-erythroid 2–related factor 2 (NRF2) target genes, and (iii) importantly, sustained activation of the AMP- activated protein kinase (AMPK pathway and associated changes in lipid metabolizing genes. This research project will utilize the GCLM KO model to expand upon our preliminary studies and investigate our working hypothesis that chronic GSH depletion induces redox activation of the AMPK pathway that serves as the central link triggering protective mechanisms that prevent AFLD. We propose to: elucidate redox-associated mechanism(s) sustaining AMPK activation in KO hepatocytes, identify AMPK catalytic subunit isoform- dependent pathways involved in ethanol-associated metabolic and stress response in KO hepatocytes, and determine the contribution of hepatic versus extrahepatic effects of GSH deficiency in modulating AMPK pathway and the protective phenotype of KO mice. The findings from these studies will provide important mechanistic information regarding key signaling and metabolic pathways involved in protection against alcohol-induced liver damage. It is anticipated that such new knowledge will reveal novel therapeutic targets for the treatment of alcohol-induced liver injury, such as AFLD. This K01 Award will allow the applicant to acquire advanced knowledge and research competency in alcohol research through an integration of interdisciplinary resources. The applicant has assembled an advisory committee composed of an outstanding group of mentors and consultants. Dr. Vasilis Vasiliou (primary mentor) is a recognized leader in the field of ethanol metabolism and toxicity. Dr. Wajahat Zafar Mehal (co-mentor) is a renowned hepatologist and liver immunologist who has extensive expertise in alcoholic and non-alcoholic fatty liver disease. Dr. Michael Harris Nathanson (co- mentor) is the Director of the Yale Liver Center and is among the world leaders in studying signaling pathways in liver disease. The program will enlist the expertise of Dr. Hongyu Zhao (consultant), who is among the world leaders in the application of statistical methods in molecular biology. Each member of the advisory committee has a formidable record of training academically successful independent scientists and they will mentor the applicant's career and academic development as well as the implementation of proposed experiments.

项目摘要 与肝硬化相关的死亡是美国的主要死亡原因之一，酒精性脂肪肝病（AFLD）陈述 这些死亡人数中有48％。尽管健康和经济影响很大，但AFLD的管理仍然是一个挑战前景 因为：（i）没有有效的诊断工具或生物标志物来评估个人对AFLD开发的易感性或 发展到更严重的临床状况，（ii）在过去的四十年中，没有开发出新的治疗实体。 该研究项目的长期目标是阐明保护肝脏免受AFLD的氧化还原相关机制 从而确定针对该疾病的潜在新型预防和/或热靶标。氧化应激在 许多途径参与AFLD的发病机理。导致氧化应激发展的主要因素是 谷胱甘肽（GSH）的耗竭，肝脏中最丰富的非蛋白质硫醇。我们使用GCLM敲除（KO）的初步研究 小鼠表明，肝GSH的缺乏率约为85％，使小鼠免受慢性乙醇诱导的脂肪变性 行政。这些小鼠也存在：（i）增强代谢乙醇和乙醛的能力，（ii）持续的氧化应激 诱导核因子2相关因子2（NRF2）靶基因，以及（iii）重要的是，AMP-持续激活 活化蛋白激酶（AMPK途径和脂质代谢基因的相关变化。该研究项目将利用 GCLM KO模型扩展了我们的初步研究，并研究了我们的工作假设，即慢性GSH耗竭会诱导 AMPK途径的氧化还原激活是触发预防AFLD的受保护机制的中央链路。我们建议 TO：阐明与氧化还原相关的机制维持KO肝细胞中AMPK激活的机制，鉴定AMPK催化亚基同工型 - KO肝细胞中与乙醇相关的代谢和应激反应涉及的依赖途径，并确定贡献 GSH缺乏症在调节AMPK途径和KO小鼠的受保护表型中的肝肝与肝外肝外影响的作用。这 这些研究的发现将提供有关关键信号传导和涉及的代谢途径的重要机械信息 防止酒精引起的肝脏损害。可以预料，这样的新知识将揭示出新的治疗靶点 治疗酒精引起的肝损伤，例如AFLD。该K01奖将允许申请人获得高级知识，并且 通过整合跨学科资源的酒精研究能力。申请人组装了 由杰出的导师和顾问组成的咨询委员会。 Vasilis Vasiliou博士（主要导师）是 公认的乙醇代谢和毒性领域的领导者。 Wajahat Zafar Mehal博士（同事）是一位著名的肝病学家和 肝免疫学家在酒精和非酒精脂肪肝病方面具有广泛的专业知识。迈克尔·哈里斯·内森（Michael Harris Nathanson）博士（共同 导师）是耶鲁大学肝脏中心的主任，是研究肝病信号通路的世界领导者。 计划将招募Hongyu Zhao（顾问）的专业知识，后者是统计应用领导者之一 分子生物学的方法。咨询委员会的每个成员都有巨大的培训记录，可以准确成功 独立科学家，他们将指导申请人的职业和学术发展以及实施 提出的实验。