A novel biological crosstalk between sumoylation and mitochondria dysfuntion in alcoholic liver disease

酒精性肝病中苏酰化和线粒体功能障碍之间的新型生物串扰

• 批准号: 10006497
• 负责人: Maria Lauda Tomasi
• 金额: $ 20.19万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-02 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10006497
• 关键词: Adenosine Triphosphate; Affect; Alcohol abuse; Alcohol consumption; Alcoholic Liver Diseases; Alcohols; Apoptosis; Apoptotic; Binding; Biological; CYP2E1 gene; Cell Death; Cell Membrane Permeability; Cell physiology; Cirrhosis; Complex; DNA Damage; DNA Repair; Data Analyses; Defect; Development; Electron Transport; Ethanol; Ethanol Metabolism; Event; Food; Generations; Genetic Transcription; Gills; Health; Heavy Drinking; Hepatocyte; Homeostasis; Inflammation; Intracellular Transport; Investigation; Kupffer Cells; Laboratories; Lead; Lipopolysaccharides; Liver; Liver Fibrosis; Liver Mitochondria; Maintenance; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Mediating; Membrane; Membrane Potentials; Metabolism; Mitochondria; Mitochondrial DNA; Mitochondrial Diseases; Mitochondrial Proteins; Modeling; Modification; Molecular; Morbidity - disease rate; Morphology; Mus; National Institute on Alcohol Abuse and Alcoholism; Necrosis; Nutrient; Organism; Oxidative Phosphorylation; Oxidative Stress; Oxygen; Oxygen Consumption; Pathway interactions; Play; Post-Translational Protein Processing; Process; Production; Proteins; Proteome; Proteomics; Reactive Oxygen Species; Regulation; Respiration; Respiratory Chain; Role; SOD2 gene; Signal Transduction; Site; Testing; Tissues; Ubiquitin; Ubiquitin-Conjugating Enzymes; alcohol effect; alcohol exposure; alcohol measurement; alcohol response; base; cell injury; cytochrome c; design; falls; fatty acid metabolism; feeding; in vivo; knock-down; liver function; microsomal ethanol-oxidizing system; mitochondrial dysfunction; mortality; mouse model; multicatalytic endopeptidase complex; novel; novel therapeutics; prevent; respiratory; therapeutic target

ABSTRACT All living organisms require exogenous foods/nutrients for producing energy in the form of Adenosine Triphosphate (ATP), which is needed for numerous cellular functions and is efficiently produced in mitochondria that are intimately involved in the generation of and defense against reactive oxygen species (ROS). An early event that occurs in response to alcohol consumption is mitochondrial dysfunction, which is evident in changes to the mitochondrial proteome, respiration defects and mitochondrial DNA (mtDNA) damage. These effects of ethanol are prominent in the liver, the major site of ethanol metabolism in the body, promoting both apoptotic and necrotic cell death and contribute to the onset or progression of alcohol-induced liver diseases (ALD), leading to hepatic fibrosis/cirrhosis and cancer. SUMOylation is a posttranslational modification that involves addition of SUMOs (small ubiquitin-like modifiers) modulating protein stability, activity and localization. Several studies have intimated a close relationship between SUMOylation and ROS. We recently demonstrated that Ubiquitin Conjugating Enzyme 9 (UBC9), the sole E2 protein required by SUMOylation machinery, is upregulated in Intragastric fed-mice liver (IE) and cirrhotic tissues. We also found that UBC9 is phosphorylated and this is correlated with high level of SUMOylation activity in lipopolysaccharides-activated Kupffer cells that leads to inflammation development. In addition, we elucidated the key function of SUMOylated microsomal Cytochrome P450 2E1 (CYP2E1) in ALD that sustains its enzymatic activity and protein stability. Much of the detailed investigation of the role of SUMOylation in ALD has been started in our laboratory. However, several important mechanistic pathways that are altered in ALD have not been investigated yet. In order to explore the role of SUMOylation in ALD, Mass Spectrometry (MS) was performed to identify SUMOylated proteins in 10-day ethanol-feeding+1 Binge ethanol (NIAAA) model, where SUMOs binding columns were used to purify SUMOylated proteins from total livers. Interestingly, we found that ethanol induces changes in SUMOylation state of several mitochondrial proteins involved in ATP synthesis (ATP5B, SOD2, CLC25A5, HSPD1, FBP1, CYCS), ATP metabolism (ATP5A1, ATP5B, ATP5C1, ATP5F1, ATP5J2, ASPA8, AK3), and mitochondria disorder (UQCRC2, PC, HMGCS2, ECHS1, COX6B1, NDUFV3, COX6C). These finding could suggest a potential role of SUMOylation in oxidative phosphorylation, electron transport chain (ETC) and respiratory control ratio may be modulating the ability of these proteins to form the complex above and/or regulating their activity. This proposal tests the novel hypothesis that ethanol induces mitochondrial dysfunction in ALD regulating the SUMOylation status of key respiratory chain proteins and explore the molecular mechanisms. Two specific aims are proposed: 1) Explore the role of SUMOylation in ethanol-modulated ETC and ATP production, 2) Examine the role of SUMOylation in ethanol-induced CYP2E1 for maintenance of mitochondrial oxygen homeostasis. If successfully completed, these studies should provide highly novel information on the role of SUMOylation in the development of ALD and may provide novel therapeutic strategies, which is of high

抽象的 所有生物都需要外源性食品/营养物质，以腺苷的形式产生能量 三磷酸（ATP），这是许多细胞功能所需的，并在线粒体中有效产生 与反应性氧（ROS）的产生和防御密切相关的。早 响应饮酒的事件是线粒体功能障碍，这在变化中很明显 对于线粒体蛋白质组，呼吸缺陷和线粒体DNA（mtDNA）损伤。这些影响 乙醇在肝脏中突出，这是体内乙醇代谢的主要部位，促进了两者 和坏死细胞死亡，并有助于酒精诱导的肝脏疾病（ALD）的发作或进展 肝纤维化/肝硬化和癌症。 sumoylation是一种翻译后修饰，涉及添加 Sumos（小泛素样修饰符）调节蛋白质稳定性，活性和定位。有几项研究 sumoylation和ROS之间存在密切的关系。我们最近证明了泛素 共轭酶9（UBC9）是Sumoylation机械所需的唯一E2蛋白，在 胃内喂养 - 肝（IE）和肝硬化组织。我们还发现UBC9是磷酸化的，这是 与脂多糖激活的库普弗细胞中高水平的Sumoylation活性相关，该细胞导致 炎症发展。此外，我们阐明了Sumoyper的微粒体细胞色素的关键功能 ALD中的P450 2E1（CYP2E1）持续其酶活性和蛋白质稳定性。大部分详细 我们的实验室已经开始研究Sumoylation在ALD中的作用。但是，有几个很重要 尚未研究ALD中改变的机械途径。为了探索 进行ALD中的sumoylation，进行质谱（MS），以鉴定10天内的sumoy蛋白 乙醇喂养+1个暴饮暴食乙醇（NIAAA）模型，其中使用Somos结合柱用于净化 来自总肝脏的Sumoypated蛋白质。有趣的是，我们发现乙醇诱导了Sumoylation的变化 参与ATP合成的几种线粒体蛋白的状态（ATP5B，SOD2，CLC25A5，HSPD1，FBP1， CYCS），ATP代谢（ATP5A1，ATP5B，ATP5C1，ATP5F1，ATP5J2，ASPA8，AK3）和线粒体 障碍（UQCRC2，PC，HMGCS2，ECHS1，COX6B1，NDUFV3，COX6C）。这些发现可能表明 Sumoylation在氧化磷酸化，电子传输链（ETC）和呼吸控制中的潜在作用 比率可能正在调节这些蛋白质形成上面的复合物和/或调节其活性的能力。 该建议检验了乙醇在ALD中诱导线粒体功能障碍的新假设 调节关键呼吸链蛋白的Sumoylation状态并探索分子 机制。提出了两个具体的目的：1）探索sumoylation在乙醇调节中的作用 等等和ATP产生，2）检查Sumoylation在乙醇诱导的CYP2E1中的作用 维持线粒体氧稳态。如果成功完成，这些研究应提供 关于Sumoylation在ALD发展中的作用的高度新颖信息，并可能提供新颖的信息 治疗策略很高