Alcohol-induced epigenetic changes in the liver genome

酒精引起的肝脏基因组表观遗传变化

基本信息

批准号：
7295781
负责人：
SAMSON T JACOB
金额：
$ 17.55万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-30 至 2008-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7295781
关键词：
2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole Acetaldehyde Acids Adenomatous Polyps Affect Alcohol abuse Alcoholic Liver Diseases Alcohols Alleles Animal Model Antibodies Biochemical Biological Markers California Characteristics Chromatin Chronic Condition Coupled CpG Islands DNA DNA Damage DNA Methylation DNA Methyltransferase DNA Methyltransferase Inhibitor DNA Modification Methylases Development Digestion Environmental Carcinogens Enzymes Epigenetic Process Ethanol Ethanol Metabolism Free Radical Scavengers Functional disorder Gel Gene Expression Gene Expression Regulation Gene Silencing Gene-Modified Genes Genetic Genome Genomic Instability Genomics Global Change Growth Hepatic Hepatocyte Hepatotoxicity Heterozygote Histone Code Histone H3 Histone H4 Histones Homocysteine Homocystine Homologous Gene Hypermethylation Immune System Diseases Incidence Injury Intervention Intestinal Neoplasms Laboratories Lead Liver Liver Cirrhosis Liver Dysfunction Malignant Neoplasms Malignant neoplasm of liver Malnutrition Mass Spectrum Analysis Measures Mediating Metabolism Methionine Methylation Microarray Analysis Mismatch Repair Mission Modification Molecular Profiling Morbidity - disease rate Mus Mutant Strains Mice N-terminal National Institute on Alcohol Abuse and Alcoholism Neuraxis Oncogenes Organ Oxidation-Reduction Oxidative Stress Pathogenesis Pathway interactions Phenotype Physiological Play Post-Translational Protein Processing Predisposition Primary carcinoma of the liver cells Protein Isoforms Proteomics Reaction Regulation Regulator Genes Relative (related person)Repetitive Sequence Research Resistance Reverse Transcriptase Polymerase Chain Reaction Role Staging Tail Testing Time Tissues Translations Tumor Suppressor Genes Universities Urea Variant Week Western Blotting Wild Type Mouse alcohol exposure chronic alcohol ingestion cocarcinogen feeding gel electrophoresis genome-wide analysis in vivo liver function metaplastic cell transformation methionine adenosyltransferase mortality promoter small molecule transmethylation tumor

项目摘要

DESCRIPTION (provided by applicant): Alcohol, a leading cause of morbidity and mortality in the world, affects many organs, most notably liver and central nervous system. An important biochemical consequence of alcohol abuse is a significant decrease in S-adenosyl methionine (SAM) level and a corresponding increase in S-adenosyl homocysteine (SAH) level. Our hypothesis is that alcohol-induced alteration in hepatic SAM and SAH levels plays a key role in modifying liver epigenome leading to altered gene expression that causes hepatic dysfunction, liver toxicity and ultimately hepatocarcinogeneis. The long term objective of this study is to advance our understanding of the role of epigenetic regulation of specific genes in mediating alcohol-mediated liver dysfunction. Altered SAM level also affects cellular redox state that influences histone modification and ultimately gene expression. The present study will test the hypothesis using mouse promoter microarray and ChlP-on-Chip analysis as well as Dnmt1 (the major DNA methyltransferase) hypomorphic mice with reduced level of Dnmt1 . The specific aims are (1) identify alcohol-induced post-translational modifications of histone H4 that occur in vivo in the livers of alcohol-fed mice by acid-urea gel electrophoresis, in-gel digestion, MALDI-TOF MS and LC/MS-MS analysis, (2) determine alterations in the DNA methylation machinery by western blot analysis and changes in the liver epigenome that occur in mice exposed to alcohol by identifying genes with altered DNA methylation status and histone post-translational modification profile using differential methylation hybridization and ChlP-on-chip analysis respectively and (3) investigate pathological changes that occur in the livers of Dnmt1 hypermorphic mice after chronic alcohol exposure and determine whether the methylation profile of genome in the livers of these mice is distinct from that of the wild type mice subjected to chronic alcohol exposure. It is anticipated that this study will identify critical epigenetic alterations that occur in alcohol-induced liver diseases. Because epigenetic changes can be reversed with relative ease by pharmacological intervention with small molecule inhibitors of DNA methyltransferase and/or histone deacetylases some of these agents could be used effectively as potential theapeutic agents in alcohol-induced liver injury. This proposal also fits well with the mission of the National Institute of Alcohol Abuse and Alcoholism on "alcohol metabolism and epigenetic effects on tissue injury".

描述（由申请人提供）：酒精是世界上发病和死亡的主要原因，影响许多器官，尤其是肝脏和中枢神经系统。酗酒的一个重要生化后果是 S-腺苷甲硫氨酸 (SAM) 水平显着降低，而 S-腺苷同型半胱氨酸 (SAH) 水平相应增加。我们的假设是，酒精引起的肝脏 SAM 和 SAH 水平的改变在改变肝脏表观基因组方面发挥着关键作用，导致基因表达改变，从而导致肝功能障碍、肝毒性并最终导致肝癌。这项研究的长期目标是加深我们对特定基因的表观遗传调控在介导酒精介导的肝功能障碍中的作用的理解。改变的 SAM 水平还会影响细胞氧化还原状态，从而影响组蛋白修饰并最终影响基因表达。本研究将使用小鼠启动子微阵列和 ChlP-on-Chip 分析以及 Dnmt1（主要 DNA 甲基转移酶）低等态小鼠（Dnmt1 水平降低）来检验该假设。具体目标是 (1) 通过酸-尿素凝胶电泳、凝胶内消化、MALDI-TOF MS 和 LC/MS 鉴定酒精喂养的小鼠肝脏中发生的酒精诱导的组蛋白 H4 翻译后修饰-MS 分析，(2) 通过蛋白质印迹分析确定 DNA 甲基化机制的变化，并通过识别 DNA 甲基化状态和组蛋白翻译后修饰改变的基因来确定暴露于酒精的小鼠中发生的肝脏表观基因组的变化分别使用差异甲基化杂交和 ChlP-on-chip 分析进行图谱分析，(3) 研究慢性酒精暴露后 Dnmt1 超态小鼠肝脏中发生的病理变化，并确定这些小鼠肝脏中基因组的甲基化图谱是否与长期暴露于酒精的野生型小鼠的结果。预计这项研究将确定酒精引起的肝病中发生的关键表观遗传改变。由于表观遗传变化可以通过 DNA 甲基转移酶和/或组蛋白脱乙酰酶小分子抑制剂的药物干预相对容易地逆转，因此其中一些药物可以有效地用作酒精性肝损伤的潜在治疗剂。这一提议也非常契合美国国家酒精滥用和酒精中毒研究所关于“酒精代谢和表观遗传对组织损伤的影响”的使命。