Alcohol-Induced Epigenetic Changes in Stem Cells

酒精引起的干细胞表观遗传变化

• 批准号: 8496655
• 负责人: LORRAINE J GUDAS
• 金额: $ 18.66万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8496655
• 关键词: ATP Citrate (pro-S)-Lyase; Abbreviations; Acetaldehyde; Acetylation; Addictive Behavior; Address; Adult; Affect; Alcohol abuse; Alcohol consumption; Alcohol dehydrogenase; Alcohols; All-Trans-Retinol; Brain; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Structure; Complex; DNA; DNA Methylation; DNA Sequence; Data; Databases; Disease; Elements; Embryo; Employee Strikes; Epigenetic Process; Ethanol; Gene Expression; Gene Expression Profiling; Gene Mutation; Genome; Heart; Hepatocyte; Histone H3; Histones; Injury; Lead; Life; Light; Liver; Liver Cirrhosis; Lysine; Metabolic; Molecular; Mus; Nucleotides; Organ; Pattern; Pharmaceutical Preparations; Polycomb; Process; Proliferating; Proteins; Rattus; Reporting; Research; Signal Pathway; Signaling Molecule; Stem cells; Sulfites; Technology; Testing; Time; Tissues; Tretinoin; Variant; Vitamin A; addiction; alcohol abuse therapy; alcohol behavior; alcohol effect; alcohol exposure; aldehyde dehydrogenases; base; cell type; chromatin immunoprecipitation; embryonic stem cell; genome-wide; genome-wide analysis; histone acetyltransferase; histone modification; insight; leukemia; medical schools; promoter; protein complex; relating to nervous system; repaired; research study; response; retinaldehyde dehydrogenase; stem; stem cell differentiation; tissue regeneration; transcriptome sequencing; tumor; tumor addiction

DESCRIPTION (provided by applicant): Alcohol consumption causes damage to multiple organs, including the liver and brain, and ethanol can affect the epigenetic state of cells by modifying DNA methylation and modifying histones. These epigenetic changes may compromise regeneration of tissues, predispose cells to become neoplastically transformed, and/or alter the function of stem cells or differentiated cells. In pilot experiments testing the effects of ethanol on murine embryonic stem (ES) cells, we observed some striking effects of ethanol and acetaldehyde, a metabolite of ethanol, on both gene expression and on histone modifications. Indeed, we already have data suggesting that alcohol influences the activity of the Polycomb Repressive Complex 2, a multi-protein complex that is a central negative epigenetic regulator of stem cell differentiation. Based on these preliminary results, our hypothesis is that alcohol and acetaldehyde induce epigenetic changes in stem cells, influencing the ability of such stem cells to undergo normal differentiation. In Aim (1) we propose to determine the global effects of alcohol and acetaldehyde on the epigenetic state of murine embryonic stem (ES) cells by performing genome-wide analysis using chromatin immunoprecipitation-sequencing (ChIP-Seq) and quantifying genome-wide changes in DNA methylation by Reduced-Representation Bi-sulfite Sequencing (RRBS). In Aim (2) we will determine how changes in the epigenetic state induced by alcohol and acetaldehyde affect gene expression and the differentiation of ES cells by performing genome-wide analysis using RNA-seq technology and by interpreting these changes in light of the changes in chromatin structure (Aim 1). We will use all-trans retinoic acid (RA), a signaling molecule that induces ES cell differentiation, as a positive control in Aim (1) since we and others have shown that RA can change the epigenetic state of ES cells. Since alcohol causes tissue injury, requiring stem cells in adult tissues to proliferate and differentiate repeatedly in an attempt to repair the damage, ou studies will lead to insights into the mechanisms by which alcohol both causes tissue injury and compromises response to tissue injury. We hypothesize that alcohol-induced epigenetic changes may underlie disorders as diverse as addictive behavior, tumor formation, and cirrhosis of the liver. Moreover, unlike genetic mutations that change the DNA sequence and are generally permanent, epigenetic changes can be reversed and therefore are reasonable targets for therapies to treat alcohol-induced tissue injury.

描述（申请人提供）：饮酒会对包括肝脏和大脑在内的多个器官造成损害，并且乙醇可以通过改变DNA甲基化和修饰组蛋白来影响细胞的表观遗传状态。这些表观遗传变化可能会损害组织的再生，使细胞易于发生肿瘤转化，和/或改变干细胞或分化细胞的功能。在测试乙醇对小鼠胚胎干 (ES) 细胞影响的初步实验中，我们观察到乙醇和乙醛（乙醇的代谢物）对基因表达和组蛋白修饰有一些显着影响。事实上，我们已经有数据表明酒精会影响 Polycomb 抑制复合物 2 的活性，Polycomb 抑制复合物 2 是一种多蛋白复合物，是干细胞分化的主要负表观遗传调节因子。基于这些初步结果，我们的假设是酒精和乙醛会诱导干细胞的表观遗传变化，影响此类干细胞进行正常分化的能力。在目标（1）中，我们建议 通过使用染色质免疫沉淀测序 (ChIP-Seq) 进行全基因组分析并通过简化表征量化 DNA 甲基化的全基因组变化，确定酒精和乙醛对小鼠胚胎干 (ES) 细胞表观遗传状态的整体影响亚硫酸氢盐测序 (RRBS)。在目标 (2) 中，我们将使用 RNA-seq 技术进行全基因组分析，并根据酒精和乙醛诱导的表观遗传状态的变化来解释这些变化如何影响基因表达和 ES 细胞的分化。染色质结构（目标 1）。我们将使用全反式视黄酸 (RA)（一种诱导 ES 细胞分化的信号分子）作为 Aim (1) 中的阳性对照，因为我们和其他人已经证明 RA 可以改变 ES 细胞的表观遗传状态。由于酒精会导致组织损伤，需要成体组织中的干细胞反复增殖和分化才能修复损伤，因此我们的研究将深入了解酒精导致组织损伤和损害对组织损伤的反应的机制。我们假设酒精引起的表观遗传变化可能是成瘾行为、肿瘤形成和肝硬化等多种疾病的基础。此外，与改变 DNA 序列且通常是永久性的基因突变不同，表观遗传变化可以逆转，因此是治疗酒精引起的组织损伤的合理目标。