LncRNA H19 in Cholestatic Liver Diseases

LncRNA H19 在胆汁淤积性肝病中的作用

• 批准号: 9750721
• 负责人: PHILLIP B HYLEMON
• 金额: $ 53.65万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-25 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9750721
• 关键词: Apical; Bile Acids; Bile Duct Epithelium; Biliary; Biological Process; Carbon Tetrachloride; Carcinoma; Cell Proliferation Regulation; Characteristics; Cholestasis; Chronic; Code; Data; Disease; Ductal Epithelial Cell; Epithelial; Exons; Fibrosis; Functional disorder; Gene Expression; Genes; Genetic Transcription; Goals; Growth; H19 gene; Hepatic; Hepatobiliary; Hepatocyte; Homeostasis; Human; Inflammation; Injury; Knock-out; Life Cycle Stages; Ligation; Link; Liver; Liver diseases; Maintenance; Mediating; Messenger RNA; Metastatic Neoplasm to the Liver; Minority; Multi-Drug Resistance; Mus; Output; Pathogenesis; Patients; Physiological; Play; Primary biliary cirrhosis; Proteins; RNA Splicing; Regulation; Regulator Genes; Reporting; Role; Sodium; Sphingosine-1-Phosphate Receptor; Structure; Taurine Cholate; Taurocholate Sodium; Testing; Tissues; Translations; Untranslated RNA; Up-Regulation; base; bile acid transporter; bile duct; cell type; cholangiocyte; cholestatic injury; cholestatic liver disease; chromatin modification; differential expression; effective therapy; human disease; imprint; knock-down; liver development; liver injury; mammalian genome; mouse model; novel; novel strategies; novel therapeutics; polypeptide; prevent; primary sclerosing cholangitis; Apical; Bile Acids; Bile Duct Epithelium; Biliary; Biological Process; Carbon Tetrachloride; Carcinoma; Cell Proliferation Regulation; Characteristics; Cholestasis; Chronic; Code; Data; Disease; Ductal Epithelial Cell; Epithelial; Exons; Fibrosis; Functional disorder; Gene Expression; Genes; Genetic Transcription; Goals; Growth; H19 gene; Hepatic; Hepatobiliary; Hepatocyte; Homeostasis; Human; Inflammation; Injury; Knock-out; Life Cycle Stages; Ligation; Link; Liver; Liver diseases; Maintenance; Mediating; Messenger RNA; Metastatic Neoplasm to the Liver; Minority; Multi-Drug Resistance; Mus; Output; Pathogenesis; Patients; Physiological; Play; Primary biliary cirrhosis; Proteins; RNA Splicing; Regulation; Regulator Genes; Reporting; Role; Sodium; Sphingosine-1-Phosphate Receptor; Structure; Taurine Cholate; Taurocholate Sodium; Testing; Tissues; Translations; Untranslated RNA; Up-Regulation; base; bile acid transporter; bile duct; cell type; cholangiocyte; cholestatic injury; cholestatic liver disease; chromatin modification; differential expression; effective therapy; human disease; imprint; knock-down; liver development; liver injury; mammalian genome; mouse model; novel; novel strategies; novel therapeutics; polypeptide; prevent; primary sclerosing cholangitis

Cholangiopathies, such as primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC), are characterized by damage and dysfunction of bile duct epithelial cells (cholangiocytes). Recently, long noncoding RNAs (lncRNAs) have been identified as a novel class of master regulators of gene expression and are linked to many fundamental biological processes and various human diseases including various liver diseases. However, little is known regarding the role of lncRNAs in the regulation of cholangiocyte function and pathogenesis of hepatobiliary diseases. The overall goal of the current application is to identify the roles and mechanisms of lncRNAs in biliary dysfunction under cholestatic conditions and to create a fundamental base for developing novel therapeutic strategies for cholangiopathies. The expression of lncRNAs is tissue-, cell type- and differentiation stage-specific. LncRNA H19 is the first identified imprinted lncRNA and is highly conserved across lineages. It has been reported that H19 is the most strongly differentially expressed lncRNA during liver development and has been linked to hepatic metastases from a range of human carcinomas and cholestatic liver injury. However, the regulatory role of H19 in cholangiocyte pathophysiology remains unknown and is the focus of the current application. Our most recent studies discovered that H19 is highly expressed in cholangiocytes, but not in hepatocytes under physiological conditions and hepatic H19 expression levels are correlated with upregulation of S1PR2 and cholestatic liver injury in the multi-drug resistance 2 knockout (Mdr2-/-) mouse, a well- established mouse model of PSC and PSC patient liver. Our preliminary data further showed that 1) BDL significantly up-regulated H19 and down-regulated the apical sodium bile acid transporter (ASBT) and sodium/taurocholate co-transporting polypeptide (NTCP); 2) BDL-induced cholestatic liver injury was markedly reduced in H19ΔExon1/+ mouse; 3) Knocking down H19 not only significantly reduced taurocholate (TCA)- induced expression of fibrotic genes and S1PR2 in cholangiocytes, but also markedly upregulated hepatic small heterodimer partner (SHP) expression and reduced cholestatic injury in Mdr2-/- mice; 4) Hepatic H19 level was also significant upregulated in the carbon tetrachloride (CCl4)-induced cholestatic liver injury mouse model. Based on these observations, we HYPOTHESIZE that lncRNA H19 plays an important role in the regulation of hepatobiliary epithelial function by disruption of hepatic bile acid homeostasis. Two specific aims are proposed to test this hypothesis. 1) To define the role of lncRNA H19 in the regulation of bile acid-mediated cholangiocyte growth and remodeling during cholestatic liver injury; 2) To identify the mechanisms by which bile acids upregulate lncRNA H19 in cholestatic conditions. Completion of the proposed studies will make a significant conceptual advance by linking the lncRNA H19-mediated regulation of biliary epithelial function with cholestatic biliary injury in patients with cholangiopathies and will provide a translational mechanism for how bile acids and lncRNA H19 mediate hepatobiliary fibrosis.

胆管疾病，例如原发性胆汁肝硬化（PBC）和原发性硬化性胆管炎（PSC） 以胆管上皮细胞（胆管细胞）的损伤和功能障碍为特征。最近，长期不编码 RNA（LNCRNA）已被确定为基因表达的一类新型主调节剂，并被联系起来 对于许多基本的生物学过程和包括各种肝脏疾病在内的各种人类疾病。 然而，关于lncRNA在胆管细胞功能调节和 肝疾病的发病机理。当前应用程序的总体目标是确定角色和 胆汁淤积条件下胆道功能障碍中LNCRNA的机制，并为 为胆管疾病制定新的治疗策略。 LNCRNA的表达是组织 - 细胞类型 - 和分化阶段特定的。 lncRNA H19是第一个鉴定出的印迹lncRNA，是高度保守的 跨血统。据报道，H19是肝脏期间最有区别的LNCRNA 开发并与来自人类癌和胆汁淤积的肝脏的肝转移有关 受伤。但是，H19在胆管细胞病理生理学中的调节作用仍然未知，是重点 当前申请。我们最近的研究发现，H19在胆管细胞中高度表达， 但是在物理条件下的肝细胞和肝H19表达水平与 多药耐药性2敲除（MDR2 - / - ）小鼠的S1PR2和胆固醇肝损伤的上调 建立的PSC和PSC患者肝脏的小鼠模型。我们的初步数据进一步表明1）BDL 明显上调的H19并下调了胆汁酸钠转运蛋白（ASBT）和 钠/牛梨酸酯共转运多肽（NTCP）; 2）BDL诱导的胆汁淤积性肝损伤显着 在H19ΔEXON1/+小鼠中降低； 3）击倒H19不仅显着降低了牛角胆酸盐（TCA） - 在胆管细胞中诱导的纤维化基因和S1PR2的表达，但也明显更新了肝小 异二聚体伴侣（SHP）表达和MDR2 - / - 小鼠中的胆固醇损伤降低； 4）肝H19水平是 在四氯化碳（CCL4）诱导的胆固醇肝损伤小鼠模型中也有明显的上调。 基于这些观察结果，我们假设lncRNA H19在调节中起重要作用 肝胆酸稳态的破坏，肝胆上皮功能。两个具体目标是 提议检验这一假设。 1）定义lncRNA H19在调节胆汁酸介导的作用 胆汁淤积性肝损伤期间的胆管细胞生长和重塑； 2）通过 胆汁酸会在胆固醇条件下上调lncRNA H19。拟议研究的完成将 通过连接lncRNA H19介导的胆道上皮调节来取得重大概念进步 胆管病患者的胆固性胆道损伤功能，并将提供转化机制 胆汁酸和LNCRNA H19如何介导肝纤维纤维化。