Role of aberrant organelle stress responses in alcohol-induced liver injury

异常细胞器应激反应在酒精性肝损伤中的作用

• 批准号: 8127681
• 负责人: CHENG JI
• 金额: $ 36.61万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-15 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8127681
• 关键词: Acetylation; Alcohol consumption; Alcoholic Liver Diseases; Alcohols; Animal Feed; Animal Model; Apoptosis; Biological Assay; CREB1 gene; Cell Death; Cessation of life; Chronic; Cirrhosis; Complex; DNA Polymerase II; Data; EP300 gene; Environmental Risk Factor; Epigenetic Process; Evolution; Fatty Liver; Fibrosis; Functional disorder; General Transcription Factors; Genes; Goals; Hematoxylin and Eosin Staining Method; Hepatic; Hepatocyte; In Situ Nick-End Labeling; Injury; Injury to Liver; Intracellular Accumulation of Lipids; Intrinsic factor; Knockout Mice; Lead; Life; Link; Lipids; Liver; Liver diseases; Measures; Methylation; Mitochondria; Mus; Organelles; Oxidative Stress; Pathogenesis; Pathway interactions; Plasmids; Process; Regulation; Research; Research Personnel; Role; Serum; Staining method; Stains; Steatohepatitis; Stress; System; Testing; Tetanus Helper Peptide; Transgenic Mice; Transplantation; United States; Work; advanced system; biological adaptation to stress; calreticulin; cell injury; chronic alcohol ingestion; feeding; in vivo; injury and repair; knockout animal; mitochondrial dysfunction; novel strategies; oil red O; prevent; promoter; public health relevance; recombinase; response; transcription factor; Acetylation; Alcohol consumption; Alcoholic Liver Diseases; Alcohols; Animal Feed; Animal Model; Apoptosis; Biological Assay; CREB1 gene; Cell Death; Cessation of life; Chronic; Cirrhosis; Complex; DNA Polymerase II; Data; EP300 gene; Environmental Risk Factor; Epigenetic Process; Evolution; Fatty Liver; Fibrosis; Functional disorder; General Transcription Factors; Genes; Goals; Hematoxylin and Eosin Staining Method; Hepatic; Hepatocyte; In Situ Nick-End Labeling; Injury; Injury to Liver; Intracellular Accumulation of Lipids; Intrinsic factor; Knockout Mice; Lead; Life; Link; Lipids; Liver; Liver diseases; Measures; Methylation; Mitochondria; Mus; Organelles; Oxidative Stress; Pathogenesis; Pathway interactions; Plasmids; Process; Regulation; Research; Research Personnel; Role; Serum; Staining method; Stains; Steatohepatitis; Stress; System; Testing; Tetanus Helper Peptide; Transgenic Mice; Transplantation; United States; Work; advanced system; biological adaptation to stress; calreticulin; cell injury; chronic alcohol ingestion; feeding; in vivo; injury and repair; knockout animal; mitochondrial dysfunction; novel strategies; oil red O; prevent; promoter; public health relevance; recombinase; response; transcription factor

DESCRIPTION (provided by applicant): The pathogenesis of alcohol-induced liver injury is very complex and undoubtedly involves the interplay of multiple mechanisms and pathways, dysfunction of cellular organelles, and interactions of intrinsic and environmental factors. The chronic feeding of alcohol leads to even more complexity as these processes of intracellular and intercellular injury and repair progress. We have focused our research on role of mitochondrial and ER stress responses in the evolution of alcohol-induced liver disease. We have obtained considerable data indicating that both mitochondria and ER are damaged by chronic alcohol consumption and that prolonged mitochondrial and ER stress responses contribute to liver injury including hepatic necroinflammation and cell death and severe fatty liver leading to fibrosis and cirrhosis. How alcohol regulates genetically and epigenetically the stress response genes of the two organelles is not known. To define the mechanisms by which alcohol consumption derails the mitochondrial and ER protective responses into injury promoting processes, we hypothesize that alcohol causes aberrant recruitment of transcription factors and epigenetic changes on the mitochondrial and ER stress response gene promoters which lead to liver injury. We propose to test the possibilities. The specific aims are: 1. Using quantitative PCR (qPCR) and ChIP assays, we will study the recruitment of promoter specific transcription factors (XBP-1, CHOP, ATF6, ATF4, CREB, TORC3, PGC-1a), general transcription factors (Pol II, NF-Y, NFR, Sp1, TBP and p300) and epigenetic marks (methylation of H3-K4 and H3-K79 and acetylation of H3 and H4) in promoters of mitochondrial and ER stress response genes (Grp78, Grp94, PDI, Calreticulin, HSP10, and HSP 60) by treating primary mouse hepatocytes with respective mitochondria and ER stress inducing agents; 2. We will perform in vivo study and measure the same parameters as described in Aim 1 with qPCR and ChIP assays by feeding mice alcohol compared to pair-fed control; 3. We will utilize Lox-Cre system as well as Dox-Tet advanced system to create and characterize liver specific Grp78 gene knockout mice; 4. We will determine effects of Grp78 deletion on the transcription factor recruitment and epigenetic changes on the two organelle gene promoters in the liver of mice fed alcohol, couple ER stress to mitochondrial dysfunction, and assess specific contribution of ER stress to alcohol-induced liver injury. Our overall goal is to reveal transcriptional and epigenetic abnormal regulation of the two organelle stress responses by alcohol in relation to the pathogenesis of alcoholic liver disease. We anticipate that this work will lead to new approaches to prevent or treat alcoholic liver disease and will be widely applicable in other types of liver disease. PUBLIC HEALTH RELEVANCE: Chronic excessive alcohol use leads to severe fatty liver and injury and is a leading cause of liver-related death and transplantation in the United States. We have identified links between liver disease and prolonged mitochondrial and ER stress responses caused by alcohol in animal models. This research is to understand how alcohol promotes injury processes in the two organelles and how organelle damages contribute to liver disease which will open new avenues for preventing and treating liver disease.

描述（由申请人提供）：酒精引起的肝损伤的发病机理非常复杂，无疑涉及多种机制和途径的相互作用，细胞器细胞器的功能障碍以及内在和环境因素的相互作用。随着这些细胞内和细胞间损伤和修复进展的这些过程，酒精的慢性喂养更加复杂。我们将研究重点放在线粒体和ER应力反应在酒精诱导的肝病演变中的作用。我们获得了相当大的数据，表明线粒体和ER因长期饮酒而受到损害，长时间的线粒体和ER应激反应有助于肝损伤，包括肝坏死症，细胞死亡以及严重的脂肪肝，导致纤维化和肝硬化。酒精如何在遗传和表观遗传上调节两个细胞器的应激反应基因。为了确定酒精消耗使线粒体和ER保护性反应使促进损伤过程的机制使酒精导致转录因子的异常募集以及线粒体和ER应激反应基因促进子的异常募集，从而导致肝损伤。我们建议测试可能性。 The specific aims are: 1. Using quantitative PCR (qPCR) and ChIP assays, we will study the recruitment of promoter specific transcription factors (XBP-1, CHOP, ATF6, ATF4, CREB, TORC3, PGC-1a), general transcription factors (Pol II, NF-Y, NFR, Sp1, TBP and p300) and epigenetic marks (methylation of H3-K4 and H3-K79和H3和H4）在线粒体和ER应激反应基因启动子中的H3-K79（GRP78，GRP94，PDI，Calreticulin，Hsp10和HSP 60）通过用相应的线粒体和ER应激诱导的抗原型的小鼠肝细胞来治疗原发性小鼠肝细胞，通过治疗原代小鼠肝细胞； 2。我们将在体内研究并测量与QPCR和CHIP测定AIM 1中所述的相同参数，与配对对照相比，通过喂养小鼠酒精。 3。我们将利用Lox-Cre系统以及DOX-TET高级系统来创建和表征肝脏特异性GRP78基因敲除小鼠； 4。我们将确定GRP78缺失对小鼠肝脏中两个细胞器基因启动子的转录因子募集和表观遗传变化的影响，喂养酒精的肝脏，对线粒体功能障碍的伴侣ER应激，并评估ER应激对酒精诱导的肝损伤的特定贡献。我们的总体目标是揭示与酒精性肝病发病机理有关的两种细胞器应激反应的转录和表观遗传异常调节。我们预计这项工作将导致预防或治疗酒精性肝病的新方法，并将广泛适用于其他类型的肝病。 公共卫生相关性：长期过度饮酒会导致严重的脂肪肝和受伤，是美国与肝有关的死亡和移植的主要原因。我们已经确定了肝病与长时间的线粒体和ER应力反应之间的联系，这是动物模型中酒精引起的。这项研究是了解酒精如何促进两个细胞器中的损伤过程，以及细胞器损害如何促进肝病，这将为预防和治疗肝病开放新的途径。