Mitochondrial Acetylation and Acetylome Dynamics in Alcoholic Liver Disease assessed with Heavy Water

用重水评估酒精性肝病中的线粒体乙酰化和乙酰组动力学

• 批准号: 10706527
• 负责人: Takhar Kasumov
• 金额: $ 18.79万
• 依托单位: NORTHEAST OHIO MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10706527
• 关键词: Acetylation; Affect; Alcohol consumption; Alcoholic Liver Diseases; Alcohols; Algorithms; Biochemical; Biological Assay; Brain; Cells; Cessation of life; Chronic; Cirrhosis; Citric Acid Cycle; Collaborations; Computer software; Consumption; Coupled; Cues; Cultured Cells; Data; Deacetylation; Deuterium Oxide; Diet; Disease; Electron Transport; Energy Metabolism; Enzymes; Food; Fostering; Genetic; Health; Hepatic; Histone Acetylation; Histones; Impairment; Ions; Isotope Labeling; Kinetics; Label; Letters; Link; Liquid Chromatography; Liquid substance; Liver; Liver Mitochondria; Liver diseases; Long-Term Effects; Lysine; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Metabolism; Methods; Mitochondria; Mitochondrial Proteins; Mus; Ohio; Oxidative Stress; Oxidoreductase; Patients; Peptide Fragments; Peptides; Play; Primary carcinoma of the liver cells; Process; Production; Protein Acetylation; Proteins; Proteome; Proteomics; Rattus; Regulation; Research; Resolution; Respiration; Respiratory Process; Risk Factors; Role; Side; Sirtuins; Site; Stress; Testing; Tracer; United States; Vertebral column; alcohol exposure; antioxidant enzyme; chronic alcohol ingestion; cofactor; enzyme activity; experimental study; fatty acid oxidation; in vivo; lipid metabolism; liver injury; mitochondrial dysfunction; mitochondrial metabolism; mouse model; novel; oxidation; pharmacologic; postnatal; problem drinker; proteostasis; stable isotope; stoichiometry; tandem mass spectrometry; targeted treatment; therapeutic target; tool

Summary Alcoholic liver disease (ALD) is a major cause of liver-related death. Alcohol intake is associated with hyperacetylation of hepatic mitochondrial proteins and impaired mitochondrial function. However, the significance of mitochondrial acetylation has been debated because of the low stoichiometry of acetylation. We propose that changes in acetylome dynamics, rather than levels, are the determinant of mitochondrial function. The rationale is that hepatic mitochondria may respond to alcohol-induced stress via acetyl-transfer dependent enzymatic inhibition (short-term regulation) and/or the acetyl-transfer independent regulation of protein stability (long-term regulation). To assess the impact of these acetylation-mediated changes, we will quantify mitochondrial acetylome dynamics in ALD mice liver in vivo using a stable isotope-resolved high- resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method relies on our technical advancement that dissects the isotope-labeling of acetyl moiety and peptide backbone. Acetylation turnover is determined based on labeling of acetyl moiety. The effect of acetylation on protein stability is assessed by comparisons of the half-lives of the intact native peptide and acetylated peptide fragments without acetyl moiety. We will use this method and genetic and pharmacological tools to study the role of altered acetylome dynamics in ALD mice livers. Experiments will be performed in collaboration with The Northern Ohio Alcohol Center (see LOS of Dr. Nagy) using an established mouse model of ALD induced with a Lieber-DeCarli diet that results in hepatic hyperacetylation and mitochondrial dysfunction. Aim 1 will determine if alcohol-induced acetylation alters the stability of hepatic mitochondrial proteins and mitochondrial respiration. Aim 2 will assess the impact of alcohol consumption on the acetylation turnover of mitochondrial proteins and the consequence of altered acetyl transfer on regulations of metabolic and antioxidant enzymes. Our novel tools and collaborative expertise will enable us to investigate the mechanisms of alcohol-induced mitochondrial dysfunction. Completion of this project will uncover a novel role of altered acetylation dynamics in the alcoholic liver and help to identify acetylation as a therapeutic target for the treatment of ALD.

概括 酒精性肝病（ALD）是肝脏相关死亡的主要原因。酒精摄入量与 肝线粒体蛋白的过度乙酰化和线粒体功能受损。然而， 由于乙酰化的化学计量较低，线粒体乙酰化的重要性一直存在争议。我们 提出乙酰组动力学的变化而不是水平的变化是线粒体的决定因素 功能。基本原理是肝线粒体可能通过乙酰转移对酒精引起的应激做出反应 依赖性酶抑制（短期调节）和/或乙酰基转移独立调节 蛋白质稳定性（长期调节）。为了评估这些乙酰化介导的变化的影响，我们将 使用稳定同位素高分辨率定量 ALD 小鼠肝脏中的线粒体乙酰组动力学 分辨率液相色谱-串联质谱 (LC-MS/MS)。这个方法依赖于我们的技术 剖析乙酰基部分和肽骨架的同位素标记的进展。乙酰化周转率为 根据乙酰基部分的标记确定。乙酰化对蛋白质稳定性的影响通过以下方式评估 完整天然肽和不含乙酰基部分的乙酰化肽片段的半衰期的比较。 我们将使用这种方法以及遗传和药理学工具来研究乙酰组动力学改变的作用 在 ALD 小鼠肝脏中。实验将与北俄亥俄州酒精中心合作进行（参见 Nagy 博士的 LOS）使用已建立的 ALD 小鼠模型，该模型采用 Lieber-DeCarli 饮食诱导，结果 肝脏过度乙酰化和线粒体功能障碍。目标 1 将确定酒精是否诱导乙酰化 改变肝线粒体蛋白和线粒体呼吸的稳定性。目标 2 将评估影响 饮酒对线粒体蛋白乙酰化转换的影响及其改变的后果 乙酰转移对代谢和抗氧化酶的调节。 我们的新颖工具和协作专业知识将使我们能够研究酒精诱发的机制 线粒体功能障碍。该项目的完成将揭示乙酰化动力学改变的新作用 酒精肝并有助于确定乙酰化作为 ALD 的治疗靶点。

项目成果

Relationship between hepatic and mitochondrial ceramides: a novel in vivo method to track ceramide synthesis.

• DOI: 10.1016/j.jlr.2023.100366
• 发表时间: 2023-05
• 期刊: JOURNAL OF LIPID RESEARCH
• 影响因子: 6.5
• 作者: Mucinski, Justine M;McCaffrey, Jonas M;Rector, R Scott;Kasumov, Takhar;Parks, Elizabeth J
• 通讯作者: Parks, Elizabeth J