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.

总结

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