The MicroRNA Network and the Deregulation of Liver Regeneration by Ethanol

MicroRNA 网络和乙醇对肝脏再生的失调

• 批准号: 8730532
• 负责人: Joannes B Hoek
• 金额: $ 17.85万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-05 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8730532
• 关键词: Accounting; Acute; Alcohol consumption; Alcoholic Liver Diseases; Alcoholic liver damage; Alcohols; Animal Feed; Animals; Cell Communication; Cell Cycle Progression; Cell Proliferation; Cells; Chronic; Control Animal; Data; Endothelial Cells; Ethanol; Ethanol dependence; Excision; Future; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Gene Targeting; Genes; Goals; Hepatic Mass; Hepatic Stellate Cell; Hepatocyte; Immunoprecipitation; Individual; Inhibition of Cell Proliferation; Kupffer Cells; Liver; Liver Regeneration; Mechanics; Mediating; Mediator of activation protein; Messenger RNA; MicroRNAs; Modeling; Molecular Profiling; Natural regeneration; Nature; Operative Surgical Procedures; Partial Hepatectomy; Pattern; Play; Population; Primary carcinoma of the liver cells; Process; Protocols documentation; Publishing; Rattus; Recovery; Relative (related person); Reporting; Resolution; Role; S Phase; Signal Transduction; Staging; Technology; Testing; Time; Tissues; Transcription Regulatory Protein; Up-Regulation; Viral; alcohol effect; alcohol exposure; base; cell type; crosslink; digital; feeding; in vivo; interest; laser capture microdissection; liver cell proliferation; liver repair; locked nucleic acid; mouse model; nano-string; novel; programs; public health relevance; regenerative; repaired; research study; response; restoration; stellate cell; tissue repair; transcriptome sequencing

DESCRIPTION (provided by applicant): Liver regeneration is a clinically important tissue repair mechanism that facilitates the recovery of liver mass after acute damage through proliferation of fully differentiated hepatocytes. Chronic ethanol intake interferes with the process of liver regeneration, which may contribute to the progression of alcoholic liver disease. We and others have found that chronic ethanol feeding alters the pattern of microRNA changes occurring in the remnant liver after partial hepatectomy (PHx) in the rat. Remarkably, our studies demonstrated that a presumed pro-proliferative microRNA species, miR-21, was increased in the remnant liver in ethanol-fed rats under conditions where hepatocyte proliferation was severely inhibited. Moreover, inhibition of miR-21 by the administration of a locked nucleic acid (LNA) antagonist of miR-21 (AM21) could overcome the ethanol- induced suppression of liver cell proliferation after PHx, suggesting that miR-21 contributes to ethanol- dependent inhibition of regeneration. A gene expression profiling analysis of the remnant liver demonstrated that a cluster of pro-fibrogenic hepatic stellate cell (HSC) markers was upregulated in ethanol-fed rats after PHX, but the upregulation of this cluster was suppressed in AM21-treated animals, indicating that HSC activation may play a role in the ethanol suppression of liver regeneration. Interestingly, inhibition of miR-21 by AM21 caused a concomitant upregulation of a cluster of other microRNAs, suggesting the involvement of a network of regulatory microRNAs mediating the recovery of regeneration in ethanol-treated animals. We propose to analyze the cell-type specific processes in the regenerating liver that are targeted by this regulatory microRNA network. The following specific aims are proposed: 1) To evaluate the role of miR-21 and its interacting miRNA regulatory network as cell type specific mediators of the suppressive effect of ethanol on the regenerative response to PHx. This will involve identifying miR-21 targets by HITS-CLIP analysis and analyzing populations of parenchymal cells, stellate cells and other cell types obtained from the remnant tissue by laser capture microdissection (LCM) for microRNA changes and corresponding changes in gene expression; 2) To evaluate the network roles of microRNA species that show parallel or anti-parallel changes relative to miR-21 in the response to PHx in AM21-treated animals and assessing their role in the recovery from ethanol inhibition of cell proliferation. These exploratory studies will guide a future in-depth analyses of the natur of the alcohol-induced deregulation of the microRNA network in liver regeneration with the long-term potential of identifying novel targets for therapy of alcoholic liver disease.

描述（由申请人提供）：肝脏再生是一种临床上重要的组织修复机制，可通过扩散完全分化的肝细胞来促进急性损伤后肝脏肿块的恢复。慢性乙醇摄入会干扰肝脏再生过程，这可能导致酒精性肝病的发展。我们和其他人发现，慢性乙醇的饮食改变了大鼠部分肝切除术（PHX）后残留肝脏发生的microRNA变化的模式。值得注意的是，我们的研究表明，在严重抑制肝细胞增殖的条件下，在乙醇喂养大鼠的残余肝脏中，假定的促增殖性microRNA物种miR-21增加了。此外，通过给予miR-21（AM21）的锁定核酸（LNA）拮抗剂对miR-21的抑制可以克服乙醇诱导的PHX后肝细胞增殖的抑制，这表明miR-21有助于乙醇依赖于乙醇依赖性的再生。对残留肝脏的基因表达分析分析表明，PHX后，在乙醇喂养的大鼠中，一组促纤维肝星状细胞（HSC）标记簇上调，但是该簇的上调在AM21治疗的动物中受到抑制，表明HSC激活在乙醇中可以在乙醇乙醇中作用乙醇乙醇果皮的作用。有趣的是，AM21对miR-21的抑制作用引起了其他microRNA簇的上调，这表明介导乙醇治疗动物再生的调节性microRNA网络参与。我们建议分析该调节性microRNA网络针对的再生肝脏中细胞类型的特定过程。提出了以下具体目的：1）评估miR-21及其相互作用的miRNA调节网络作为细胞类型的特定介质的作用，这些乙醇对乙醇对PHX的再生反应的抑制作用。这将涉及通过击中clip分析并分析实质细胞，星状细胞和其他细胞类型的种群来鉴定miR-21靶标，这是通过激光捕获微分解区（LCM）从残余组织中获得的，以实现microRNA的变化和基因表达的相应变化； 2）评估MicroRNA物种的网络作用，这些网络作用在AM21处理的动物中对PHX的反应中显示出平行或反行的变化，并评估其在细胞增殖乙醇抑制中恢复中的作用。这些探索性研究将指导对肝脏再生中酒精引起的microRNA网络放松管制的自然的未来深入分析，具有鉴定用于酒精性肝病治疗的新靶标的长期潜力。