Functional characters of non-coding RNAs in alcoholic liver injury

非编码RNA在酒精性肝损伤中的功能特征

• 批准号: 8461464
• 负责人: FANYIN MENG
• 金额: --
• 依托单位: OLIN TEAGUE VETERANS CENTER
• 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 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8461464
• 关键词: Accounting; Affect; Alcohol abuse; Alcoholic Fatty Liver; Alcoholic Liver Cirrhosis; Alcoholic Liver Diseases; Alcoholic beverage heavy drinker; Alcohols; Animal Model; Animals; Apoptosis; Area; Attention; Biochemical; Bioinformatics; Biological Assay; Biology; Cell Line; Cell Survival; Cell Therapy; Cells; Cellular biology; Cessation of life; Cirrhosis; Clinical; Code; Complex; Country; DNA; DNA Methylation; DNA Methyltransferase; DNA Modification Methylases; DNA biosynthesis; Data; Development; Disease; Disease Marker; Disease Progression; Endotoxins; Enzymes; Epigenetic Process; Epithelial Cells; Ethanol; Ethanol dependence; Family; Fatty Liver; Fibrosis; Functional RNA; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Health; Hepatic; Hepatic Fibrogenesis; Hepatic Stellate Cell; Hepatitis; Hepatobiliary; Hepatocyte; Human; In Vitro; Inflammation; Inflammatory; Infusion procedures; Injury; Investigation; Knowledge; Lead; Learning; Lipopolysaccharides; Liver; Liver Cirrhosis; Liver Fibrosis; Liver diseases; Location; Luciferases; Mediating; Mediator of activation protein; Methylation; MicroRNAs; Molecular; Molecular Biology; Necrosis; Organ; Oxidative Stress; Pathogenesis; Pathway interactions; Phenotype; Physiological; Play; Population; Primary carcinoma of the liver cells; Process; Property; Protein Biosynthesis; Proteins; Proteome; Public Health; RNA; Reactive Oxygen Species; Regulation; Regulator Genes; Reporter; Research; Risk; Role; Secondary to; Shapes; Signal Transduction; Signaling Molecule; Specimen; Steatohepatitis; TNF gene; Techniques; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Tissues; Transcript; Translating; Translations; United States; Up-Regulation; Veterans; Western Blotting; Work; addiction; alcohol effect; alcohol research; base; cell motility; cell type; cholangiocyte; chronic alcohol ingestion; chronic liver disease; cytokine; cytotoxic; drug of abuse; imprint; in vivo; insight; intercellular communication; interest; liver injury; mouse model; novel; novel therapeutic intervention; problem drinker; public health relevance; response; stellate cell; toll-like receptor 4

DESCRIPTION (provided by applicant): Project Summary Alcoholic liver diseases (ALD) -- ranging from alcoholic fatty liver to alcohol-induced liver fibrosis and cirrhosis - - account for more than 50% of all chronic liver diseases in western countries and are responsible for 5% of the deaths occurring annually in the United States including our Veteran population. Studies have shown that while all heavy drinkers display signs of hepatitis steatosis (fatty liver); only 10% to 35% of alcoholics develop hepatic inflammation, with up to 20% progressing to cirrhosis. About 15% of U.S. alcoholics eventually will develop alcoholic liver diseases, one of the leading causes of liver diseases and liver-relate death worldwide. The clinical spectrum of alcoholic liver disease includes alcoholic fatty liver, alcoholic steatohepatitis, alcoholic cirrhosis, and increased risk of hepatocellular carcinoma. The pathological mechanisms of ALD involves complex interactions between the direct effects of alcohol and its toxic metabolites on various cell types in the liver, induction of reactive oxygen species (ROS), upregulation of the inflammatory cascade, and other cell-specific effects in the liver. The field of non-coding RNA (ncRNA) molecules represents a paradigm shift in biology, away from the central dogma of biology which places RNA molecules as mere messengers between DNA and protein synthesis. However, more than 90% of the RNA molecules made by a cell are not destined to be translated into proteins. Instead, these ncRNAs can act as enzymes, signaling molecules and transcriptional factors. MicroRNAs (miRNAs) are small ncRNAs that have been recently identified as master regulators of the cellular transcriptome and proteome. miRNAs play a crucial role in shaping the differentiation and function of tissues and organs in both health and disease. Several recent studies have provided compelling evidence showing that ethanol-sensitive miRNAs are indeed regulatory master-switches. More specifically, miRNAs control the development of tolerance, a crucial component of ethanol addiction. Other drugs of abuse also target some ethanol-sensitive miRNAs suggesting that common biochemical mechanisms underlie addiction. Therefore, the knowledge of the precise mechanisms of ncRNA regulation in ALD becomes very critical to develop therapeutic interventions since the failing liver may revive if given proper support in a timely manner. Our long-term goal is to clarify the mechanisms underlying the abnormal intercellular communication of ncRNAs that lead to ALD. In this application, we propose the systematic investigation of ethanol and endotoxin (lipopolysaccharide, LPS) dependent ncRNAs as ALD markers by focusing on four specific aims: First, we will identify the functional ethanol-dependent miRNAs involved in cell survival in human hepatic stellate cells, hepatocytes and cholangiocytes. Second, we will define the role of LPS regulated miRNAs in human hepatic stellate and parenchymal cells and the related remodeling mechanisms. Third, we will evaluate the cross-talk between specific transcribed ultra conserved regions (T-UCRs) and miR- 181/let-7 families, and define the role of this interaction in ALD properties in hepatobiliary epithelial and stellate cells. Fourth, we will determine the effects of ethanol and LPS dependent ncRNAs on the progression of ALD in a mouse model of alcoholic steatohepatitis in vivo. The identification of miRNAs as an important regulator of hepatic cell survival, transformation and remodeling in vitro, as well as their upstream modulators and interaction with T-UCRs will provide insight into the involvement of altered ncRNA expression in contributing to ALD progression and test novel therapeutic approaches for ALD in animal models.

描述(由申请人提供)： 项目概述酒精性肝病(ALD)--从酒精性脂肪肝到酒精性肝纤维化和肝硬变--占西方国家所有慢性肝病的50%以上，占美国每年死亡人数的5%，包括我们的退伍军人。研究表明，尽管所有酗酒者都表现出肝炎脂肪变性(脂肪肝)的迹象，但只有10%到35%的酗酒者会出现肝脏炎症，高达20%的人会发展为肝硬变。大约15%的美国酗酒者最终会患上酒精性肝病，这是全球肝病和肝脏相关死亡的主要原因之一。酒精性肝病的临床范围包括酒精性脂肪肝、酒精性脂肪性肝炎、酒精性肝硬变和肝细胞癌风险增加。酒精性肝病的病理机制涉及酒精及其毒性代谢产物对肝脏各种细胞类型的直接作用、活性氧自由基(ROS)的诱导、炎症级联反应的上调以及其他细胞特异性效应之间的复杂相互作用。非编码RNA(NcRNA)分子领域代表了生物学中的一种范式转变，脱离了生物学的中心教条，即将RNA分子仅仅作为DNA和蛋白质合成之间的信使。然而，细胞制造的90%以上的RNA分子注定不会被翻译成蛋白质。相反，这些ncRNA可以作为酶、信号分子和转录因子发挥作用。MicroRNAs(MiRNAs)是最近被鉴定为细胞转录组和蛋白质组的主要调节因子的小的ncRNAs。在健康和疾病中，miRNAs在塑造组织和器官的分化和功能方面发挥着至关重要的作用。最近的几项研究提供了令人信服的证据，表明乙醇敏感的miRNAs确实是调节性的主开关。更具体地说，miRNAs控制着耐受性的发展，这是酒精成瘾的关键组成部分。其他滥用药物也针对一些乙醇敏感的miRNAs，这表明共同的生化机制是成瘾的基础。因此，了解ncRNA在ALD中的精确调控机制对于开发治疗干预措施非常关键，因为如果及时给予适当的支持，衰竭的肝脏可能会复苏。我们的长期目标是阐明导致ALD的ncRNAs异常细胞间通讯的潜在机制。在这一应用中，我们建议系统地研究依赖乙醇和内毒素(内毒素，LPS)的ncRNAs作为ALD标记物，主要集中在四个特定的目的：首先，我们将鉴定在人肝星状细胞、肝细胞和胆管细胞中参与细胞存活的功能性依赖乙醇的miRNAs。其次，我们将确定内毒素调节的miRNAs在人肝星状细胞和实质细胞中的作用以及相关的重塑机制。第三，我们将评估特定转录超保守区(T-UCR)和miR-181/let-7家族之间的相互作用，并确定这种相互作用在肝胆上皮细胞和星状细胞ALD特性中的作用。第四，我们将在体内确定酒精和脂多糖依赖的ncRNAs对酒精性脂肪性肝炎小鼠ALD进展的影响。在体外发现miRNAs是肝细胞存活、转化和重塑的重要调节因子，以及它们的上游调节因子和与T-UCR的相互作用将有助于深入了解ncRNA表达变化在ALD进展中的作用，并在动物模型中测试ALD的新治疗方法。