Role of Sumoylation in Alcoholic Liver Disease

苏酰化在酒精性肝病中的作用

• 批准号: 9320994
• 负责人: Maria Lauda Tomasi
• 金额: $ 16.16万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-20 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9320994
• 关键词: Adipocytes; Alcohol abuse; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alpha Cell; Animal Model; Apoptosis; Cell Line; Cell physiology; Cells; Cirrhosis; Country; DNA; DNA Repair; DNase-I Footprinting; Data; Development; Enzymes; Ethanol; Ethanol Metabolism; Fatty acid glycerol esters; Fibrosis; Genes; Genetic Transcription; Gills; Half-Life; Health; Hepatic; Hepatic Stellate Cell; Hepatocyte; Homeostasis; Human; Inflammatory Response; Infusion procedures; Injury; Intracellular Transport; Kupffer Cells; Lentivirus Vector; Lipids; Lipopolysaccharides; Liver; Liver diseases; Mass Spectrum Analysis; Messenger RNA; Minor; Modification; Molecular; Mus; Nuclear; Oxidative Stress; Pathogenesis; Pathway interactions; Play; Post-Translational Protein Processing; Primary carcinoma of the liver cells; Production; Proteins; RNA Interference; Reactive Oxygen Species; Reporting; Risk; Role; Running; SUMO-1 Protein; Signal Transduction; Site; Techniques; Technology; Testing; Triglycerides; Ubiquitin-Conjugating Enzymes; alcohol abuse therapy; base; biological adaptation to stress; cell type; cytokine; design; functional outcomes; in vivo; knock-down; macrophage; mortality; novel; novel therapeutics; prevent; promoter; protein activation; protein protein interaction; public health priorities; public health relevance; sumo1 gene; trafficking

DESCRIPTION (provided by applicant): Alcohol abuse is a leading factor in mortality from liver disease and increases the risk for a wide range of adverse health effects. The liver, as the primary site of alcohol metabolism, is a major target of injury. The spectrum of Alcoholic Liver Diseases (ALD) includes simple steatosis, alcoholic hepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. Sumoylation is a post-translational modification that modulates multiple cellular processes such as signal transduction, stress responses, cellular trafficking, protein-protein interactions, protein-DNA interactions and transcriptional activity. SUMO is comprised of four distinct proteins in humans (SUMO-1, -2, 3- and -4). Sumoylation is often increased under oxidative stress. We recently reported that ubiquitin conjugating enzyme 9 (Ubc9), the sole E2 enzyme of sumoylation, is induced in intragastric ethanol- infusion (EI) treated mice but the functional significance of this is unknown. Consistently, we found SUMO-1, -3 and Ubc9 mRNA levels are increased the livers of EI mice. Also, EI mice show an overall increase in protein sumoylation by SUMO-1 but only minor changes in sumoylation by SUMO-2/3. Ethanol treatment of primary mouse hepatocytes leads to increased reactive oxygen species (ROS) and triglyceride production. In addition, we found increased expression of Ubc9 and SUMO genes, Cyp2e1 and an overall increase in SUMO-1 protein sumoylation like in EI livers. Silencing of Ubc9 prevented ethanol-induced fat accumulation, ROS production and increased Cyp2e1 expression in primary mouse hepatocytes. In LX-2 cells (activated human hepatic stellate cells or HSCs), ethanol treatment also increased Ubc9 expression, ROS production and HSC activation markers. Blocking Ubc9 induction prevented all of these and induced apoptosis in HSCs. Finally, we found that lipopolysaccharide (LPS) and Ubc9 RNAi treatment alone increased expression of proinflammatory cytokines in RAW cells (macrophage cell line); but when LPS and Ubc9 RNAi were combined, the expression of these cytokines increased further. Interestingly, LPS treatment decreased Ubc9 protein level (mRNA level was unchanged). This proposal is testing the novel hypothesis that there is dysregulation in sumoylation that contributes to the pathogenesis of ALD in a cell-type specific manner. Three specific aims are proposed to examine: 1) the role of sumoylation in ethanol-induced changes in hepatocytes, 2) the role of sumoylation in ethanol-induced HSC activation, and 3) the role of sumoylation in Kupffer cell activation in ALD. If successfully completed, these studies should provide highly novel information on the role of sumoylation in the development of ALD and may provide novel therapeutic strategies, which is of high public health priority.

描述（由申请人提供）：酗酒是肝病死亡率的主要因素，并增加了多种不良健康影响的风险。作为酒精代谢的主要部位，肝脏是损伤的主要目标。酒精性肝疾病（ALD）的光谱包括简单的脂肪变性，酒精性肝炎，纤维化，肝硬化和肝细胞癌。 Sumoylation是一种翻译后修饰，可调节多个细胞过程，例如信号转导，应力反应，细胞运输，蛋白质 - 蛋白质相互作用，蛋白-DNA相互作用和转录活性。 Sumo由人类中的四种不同的蛋白质组成（SUMO -1，-2、3-和-4）。在氧化应激下通常会增加sumoylation。我们最近报道说，泛素结合酶9（UBC9）是sumoylation的唯一E2酶，在胃内乙醇输注（EI）处理的小鼠中诱导，但其功能意义是未知的。一致地，我们发现SUMO -1，-3和UBC9 mRNA水平增加了EI小鼠的肝脏。同样，EI小鼠通过SUMO-1显示了蛋白质Sumoylation的总体增加，但SUMO-2/3的Sumoylation仅发生了较小的变化。原代小鼠肝细胞的乙醇处理导致活性氧（ROS）和甘油三酸酯的产生增加。此外，我们发现UBC9和SUMO基因，CYP2E1的表达增加，以及EI肝中SUMO-1蛋白Sumoylation的总体增加。 UBC9的沉默阻止了乙醇诱导的脂肪积累，ROS产生和CYP2E1在原代小鼠肝细胞中的表达增加。在LX-2细胞（活化的人肝星状细胞或HSC）中，乙醇处理还增加了UBC9表达，ROS产生和HSC激活标记。阻止UBC9诱导阻止了HSC中的所有这些和诱导的凋亡。最后，我们发现单独的脂多糖（LPS）和UBC9 RNAi治疗增加了原细胞（巨噬细胞系）中促炎细胞因子的表达。但是，当合并LPS和UBC9 RNAi时，这些细胞因子的表达进一步增加。有趣的是，LPS处理降低了UBC9蛋白水平（mRNA水平没有变化）。该提议正在检验新的假设，即Sumoylation中存在失调，该假设以细胞类型的特定方式有助于ALD的发病机理。提出了三个具体目的来检查：1）Sumoylation在乙醇诱导的肝细胞变化中的作用，2）Sumoylation在乙醇诱导的HSC激活中的作用，以及3）Sumoylation在ALD中kupffer细胞活化中的作用。如果成功完成，这些研究应提供有关Sumoylation在ALD发展中作用的高度新颖信息，并可能提供新颖的治疗策略，这是高公共卫生优先事项。

项目成果

Post-translational deregulation of YAP1 is genetically controlled in rat liver cancer and determines the fate and stem-like behavior of the human disease.

• DOI: 10.18632/oncotarget.10246
• 发表时间: 2016-08-02
• 期刊: Oncotarget
• 作者: Simile MM;Latte G;Demartis MI;Brozzetti S;Calvisi DF;Porcu A;Feo CF;Seddaiu MA;Daino L;Berasain C;Tomasi ML;Avila MA;Feo F;Pascale RM
• 通讯作者: Pascale RM