Ethanol Effects on the Transcriptional Regulatory Network in Liver Regeneration

乙醇对肝脏再生转录调控网络的影响

基本信息

批准号：
9105305
负责人：
Joannes B Hoek
金额：
$ 52.13万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9105305
关键词：
Accounting Affect Alcohol consumption Alcoholic Liver Diseases Alcohols Animal Feed Animals Attention Automobile Driving Bioinformatics Categories Cell Communication Cell Cycle Cell Cycle Progression Cell Proliferation Cells Chronic Chronic Disease Classification Clinical Computer Analysis Computer Simulation Control Animal Data Defect Equilibrium Ethanol Experimental Models Extracellular Matrix Gene Expression Goals Health Hepatic Stellate Cell Hepatocyte Immunohistochemistry Impaired wound healing Individual Injury Intervention Kupffer Cells Liver Liver Regeneration Liver diseases Measures Mediating Mediator of activation protein Messenger RNA Metabolic MicroRNAs Modeling Molecular Profiling Mus Natural regeneration Partial Hepatectomy Pathway interactions Phenotype Process Proliferation Marker Rattus Recovery Regenerative response Regulatory Pathway Risk Role Signal Transduction Staging System Testing Time Tissues Up-Regulation alcohol abuse therapy alcohol effect alcohol exposure cell type chronic alcohol ingestion early onset experimental analysis feeding hepatic acinus structure in vivo inhibitor/antagonist insight interest laser capture microdissection liver cell proliferation liver repair mouse model novel paracrine repaired response tissue repair tool transcriptome transcriptomics translational approach

项目摘要

DESCRIPTION (provided by applicant): Liver regeneration is a clinically important tissue repair process in which the liver responds to injury by driving a synchronized replication of differentiated liver cells. The process involves the interplay of carefully orchestrated signals frm different cell types integrated with systemic factors to recover functional tissue mass. Liver regeneration is inhibited by chronic ethanol consumption and this impaired repair response may contribute to the risk for alcoholic liver disease. The mechanisms responsible for the onset and progression of the regenerative response and its deregulation by ethanol remain poorly characterized. In prior studies we carried out a detailed gene expression and microRNA profiling study of the regeneration response after 70% partial hepatectomy (PHx) in ethanol-fed and control rats and developed novel bioinformatics tools to highlight cell- type specific transcriptomic signatures. These studies indicate that chronic ethanol treatment affects non- parenchymal cells (NPCs) that control hepatocyte priming and replication and is associated with upregulation of a hepatic stellate cell (HSC) activation signature. We obtained evidence that miR-21 may be a driver of these differential responses. Inhibition of miR-21 in the rat in vivo by treatment with AM21, a specific miR-21 antagonist, allowed recovery of liver regeneration in ethanol-fed rats while suppressing the differential HSC activation profile. In the current application we build on these findings to analyze how different cell types contribute to the integrated tissue response using laser capture microdissection (LCM) and integrating the experimental data through a computational modeling approach to predict how regeneration is inhibited by adaptation to chronic ethanol exposure and rescued by miR-21 inhibition. (1) We will analyze the temporal regeneration dynamics in ethanol-fed and control rats and profile the differential impact of AM21 treatment to characterize the miR-21 target processes that contribute to the ethanol-mediated inhibition of liver regeneration, with a focus on the contribution of NPCs to cell cycle progression. We will further compare rat and mouse models of liver regeneration to evaluate how miR-21's interactions with cell cycle regulators differs in animals that have a different temporal response to PHx. (2) We will test the hypothesis that the adaptation to ethanol consumption alters the temporal response of different cell types in the liver, deregulating the integrated tissue response, by analyzing cell type-specific changes in the transcriptome and microRNA expression in ethanol-fed animals compared to controls by analyzing individual cells obtained using LCM. (3) We will apply a computational model of the role of cell-type specific interactions in liver regeneration to iteratively evaluate the consequences of dynamic changes in phenotype distribution. Our integrated experimental and computational analysis will yield new insights into the regulatory miRNA network driving cell phenotype distributions and cell-cell interactions, and the mechanisms through which interventions in these networks can counter ethanol-induced suppression of liver repair, with applications to translational and clinical interventions i in a broad range of liver diseases.

描述（由适用提供）：肝脏再生是一种临床上重要的组织修复过程，其中肝脏通过驱动分化肝细胞的同步复制来反应损伤。该过程涉及精心策划的信号FRM的相互作用不同的细胞类型，这些细胞类型与全身因素集成以恢复功能性组织质量。慢性乙醇消耗抑制了肝脏再生，这种受损的修复反应可能导致酒精性肝病的风险。导致再生反应发作和进展的机制及其对乙醇的放松管制的特征仍然很差。在先前的研究中，我们对乙醇喂养和控制大鼠进行了70％部分肝（PHX）后的再生反应进行了详细的基因表达和microRNA分析研究，并开发了新型的生物信息学工具，以突出细胞类型 - 类型的特定转录组签名。这些研究表明，慢性乙醇治疗会影响控制肝细胞启动和复制的非副作用细胞（NPC），并且与肝星状星状细胞（HSC）激活签名的上调有关。我们获得了MiR-21可能是这些差异反应的驱动力的证据。通过用特定的miR-21拮抗剂AM21处理，在大鼠体内抑制miR-21在体内抑制，允许在乙醇喂养的大鼠中恢复肝脏再生，同时抑制差异HSC激活谱。在当前的应用中，我们基于这些发现，以分析不同的细胞类型如何使用激光捕获显微解剖（LCM）对综合组织反应有何贡献，并通过计算模型方法通过计算模型方法整合了实验数据，以预测如何通过适应慢性乙醇暴露而抑制再生并受到miR-21抑制作用。（1）我们将分析乙醇喂养和控制大鼠中的暂时再生动力学，并介绍AM21处理的差异影响，以表征有助于乙醇介导的肝脏再生抑制的miR-21靶过程，重点是NPC对细胞周期进展的贡献。我们将进一步比较肝脏再生的大鼠和小鼠模型，以评估miR-21与细胞周期调节剂的相互作用如何在对PHX暂时反应不同的动物中的不同。（2）我们将测试以下假设：乙醇消耗的适应性改变了肝脏中不同细胞类型的临时反应，通过分析使用LCM获得的单个细胞相比，通过分析乙醇馈送动物中转录组和microRNA表达的细胞类型特异性变化，从而消除了整合的组织反应。（3）我们将应用一个计算模型，即细胞类型在肝脏再生中特异性相互作用的作用，以迭代评估表型分布中动态变化的后果。我们集成的实验和计算分析将为调节性miRNA网络驱动细胞表型和细胞 - 细胞相互作用提供新的见解，以及这些网络中的干预措施可以抵抗乙醇诱导肝修复的抑制的机制，并采用在宽范围内翻译和临床干预措施的应用。