Role of aberrant regulation of organelle stress responses in alcohol-induced live

细胞器应激反应异常调节在酒精诱导的生活中的作用

• 批准号: 7798824
• 负责人: CHENG JI
• 金额: $ 38.67万
• 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/7798824
• 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; Hepatic; Hepatocyte; In Situ Nick-End Labeling; Injury; Injury to Liver; Intracellular Accumulation of Lipids; 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（申请人提供）：酒精性肝损伤的发病机制非常复杂，无疑涉及多种机制和途径的相互作用、细胞器的功能障碍以及内在因素和环境因素的相互作用。随着细胞内和细胞间损伤和修复过程的进展，长期饮酒会导致更加复杂的情况。我们的研究重点是线粒体和内质网应激反应在酒精性肝病演变中的作用。我们获得的大量数据表明，长期饮酒会损害线粒体和内质网，并且长期的线粒体和内质网应激反应会导致肝损伤，包括肝坏死炎症和细胞死亡以及导致纤维化和肝硬化的严重脂肪肝。酒精如何从遗传和表观遗传角度调节这两种细胞器的应激反应基因尚不清楚。为了确定饮酒使线粒体和内质网保护性反应脱轨进入损伤促进过程的机制，我们假设酒精会导致转录因子的异常募集以及线粒体和内质网应激反应基因启动子的表观遗传变化，从而导致肝损伤。我们建议测试可能性。具体目标是： 1.使用定量PCR（qPCR）和ChIP检测，我们将研究启动子特异性转录因子（XBP-1、CHOP、ATF6、ATF4、CREB、TORC3、PGC-1a）、一般转录因子的招募（Pol II、NF-Y、NFR、Sp1、TBP 和 p300）和表观遗传标记（H3-K4 和 H3-K79 的甲基化以及 H3 和 H3-K79 的乙酰化） H4)通过用各自的线粒体和内质网应激诱导剂处理原代小鼠肝细胞，在线粒体和内质网应激反应基因(Grp78、Grp94、PDI、钙网蛋白、HSP10和HSP 60)的启动子中； 2. 我们将进行体内研究，并通过 qPCR 和 ChIP 检测测量与目标 1 中所述相同的参数，通过给小鼠喂食酒精与配对喂养对照进行比较； 3. 我们将利用Lox-Cre系统以及Dox-Tet先进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 缺失对饮酒小鼠肝脏中转录因子募集和两个细胞器基因启动子表观遗传变化的影响，将 ER 应激与线粒体功能障碍耦合，并评估 ER 应激对酒精诱导的肝脏的具体贡献受伤。我们的总体目标是揭示酒精对两种细胞器应激反应的转录和表观遗传异常调节与酒精性肝病发病机制的关系。我们预计这项工作将带来预防或治疗酒精性肝病的新方法，并将广泛应用于其他类型的肝病。 公共卫生相关性：长期过量饮酒会导致严重的脂肪肝和损伤，并且是美国肝脏相关死亡和移植的主要原因。我们在动物模型中发现了肝脏疾病与酒精引起的长期线粒体和内质网应激反应之间的联系。这项研究旨在了解酒精如何促进两种细胞器的损伤过程，以及细胞器损伤如何导致肝病，这将为预防和治疗肝病开辟新途径。