LncRNA H19 in Cholestatic Liver Diseases

LncRNA H19 在胆汁淤积性肝病中的作用

基本信息

批准号：
10909545
负责人：
PHILLIP B HYLEMON
金额：
$ 62.96万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-25 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10909545
关键词：
Algorithms Animal Model Bile Acids Biliary Bioinformatics Biological Process Ceramides Characteristics Cholestasis Clinical Code Data Diagnostic Disease Epidermal Growth Factor FDA approved Family member Fibrosis Functional disorder Funding Gene Expression Gene Expression Regulation Genes Genetic Transcription Goals H19 gene HMGB1 gene Hepatic Hepatic Stellate Cell High-Throughput RNA Sequencing Human Human Genome Impairment Inflammation Inflammatory Injury Kupffer Cells Ligation Liver Liver Fibrosis Liver diseases MLLT2 gene Macrophage Mediating Metabolism MicroRNAs Molecular Morbidity - disease rate Mus Pathogenicity Pathologic Processes Pathway interactions Patients Pattern Physiological Processes Play Porifera Proliferating Proteins Publishing RNA analysis Reporting Role Sampling Sequence Analysis Serum Severities Sphingolipids Sphingosine Taurocholic Acid Testing Therapeutic Tissues Untranslated RNA Up-Regulation bile duct bile formation cholangiocyte circular RNA clinical application human disease innovation liver inflammation liver injury mammalian genome mortality mouse model new therapeutic target novel diagnostics overexpression primary sclerosing cholangitis promoter receptor senescence single nucleus RNA-sequencing stellate cell transcriptome transcriptome sequencing

项目摘要

Primary sclerosing cholangitis (PSC) is the major cholestatic liver disease with high morbidity and mortality. Inflammation and fibrotic injury of the bile ducts due to impairment of bile formation or flow represent the major characteristics of PSC. The majority of the mammalian genome are non-protein-coding sequences. Less than 2% of the human genome is made up of protein-coding genes. A large fraction of the transcribed mammalian genome is noncoding RNAs (ncRNAs), including long noncoding RNAs (lncRNAs), circular RNA (circRNAs), and microRNA (miRNAs), but only a few of them have been structurally annotated. The biological functions of ncRNAs remain largely unknown. Recent advances in high-throughput RNA sequencing (RNAseq) and circRNA- specific bioinformatics algorithms have identified thousands of circRNAs with tissue-specific expression patterns. However, the relevance and function of circRNAs in disease, especially in liver diseases, remain to be determined. Our previous studies reported that the conjugated primary bile acid (CBA), taurocholic acid (TCA), activated sphingosine-1phosphate receptor 2 (S1PR2) and induced H19 expression in cholangiocytes. TCA- induced upregulation of H19 is a major promoter of cholestatic liver injury by activating inflammatory macrophages and hepatic stellate cells (HSCs). Our new preliminary data using Arraystar circRNAseq identified 33 overlapping circRNAs, which were significantly upregulated in Mdr2-/- mice and downregulated in Mdr2-/-/H19- /- mice. Among them, mmu_circRNA_26644 (circRNA-Edil3), which is encoded by epidermal growth factor-like repeats and discoidin I-like domains 3 (Edil3) gene, is the most significantly downregulated circRNA in Mdr2-/- /H19-/- mice. Sequence analysis suggested that circRNA-Edil3 may serve as a "sponge" for miRNA-215-3p, miRNA-129-5p and miRNA-3075-3p. TargetScan analysis suggested that High Mobility Group Box Protein 1 (HMGB1), AF4/FMR2 Family Member 3 (Aff3) and Edil3 are potential targets of miRNA-215-3p, miRNA-129-5p and miRNA-3075. Based on our published results and exciting new preliminary data, we HYPOTHESIZE that H19-induced upregulation of circRNA-Edil3 plays a critical role in cholestatic liver injury by sequestering miRNAs. Two specific aims are proposed to test our hypothesis. Aim 1. To examine the role of H19-induced circRNA-Edil3 in regulating cholangiocyte proliferation, senescence and hepatic inflammation using cholestatic liver injury mouse models and human PSC patient liver and serum samples. Aim 2: To identify the mechanisms by which H19-induced circRNA-Edil3 promotes cholestatic liver injury. We will use multiple newly-established approaches and animal models to test our hypothesis. Completion of these specific aims should elucidate the potential cellular/molecular mechanisms involved in lncRNA H19-mediated progression of cholestatic liver diseases and identify novel diagnostic and therapeutic targets. Since the effects of cholestasis are profound and widespread, leading to worsening liver diseases and systemic illness, the subject matter of this proposal is timely, important, and has direct clinical application.

原发性硬化性胆管炎（PSC）是高发病率和死亡率高的主要胆固性肝病。由于胆汁形成或流动的损害，胆管的炎症和纤维化损伤代表主要 PSC的特征。大多数哺乳动物基因组是非蛋白质编码序列。少于 2％的人基因组由蛋白质编码基因组成。抄录的哺乳动物很大一部分基因组是非编码RNA（NCRNA），包括长的非编码RNA（LNCRNA），圆形RNA（CIRCRNA），和microRNA（miRNA），但其中只有少数在结构上注释。生物学功能 NCRNA在很大程度上仍然未知。高通量RNA测序（RNASEQ）和CIRCRNA-的最新进展特定的生物信息学算法已经鉴定出具有组织特异性表达模式的数千种CIRCRNA。但是，circrNA在疾病中的相关性和功能，尤其是在肝病中，仍然是决定。我们以前的研究报告说，共轭原代胆汁酸（CBA），牛胆酸（TCA），激活的鞘氨氨酸1磷酸受体2（S1PR2）和胆管细胞中诱导的H19表达。 TCA- H19诱导的上调是通过激活炎症的主要促进剂巨噬细胞和肝星细胞（HSC）。我们使用Arraystar circrnaseq确定的新的初步数据 33重叠的CIRCRNA，在MDR2 - / - 小鼠中显着上调，并在MDR2 - / - /H19-中下调 / - 老鼠。其中，mmu_circrna_26644（circrna-edil3），由表皮生长因子样本编码重复序列和盘状蛋白I样结构域3（EDIL3）基因是MDR2 - / - 中最显着下调的circrna。 /H19 - / - 小鼠。序列分析表明，Circrna-Edil3可以用作miRNA-215-3p的“海绵”， miRNA-129-5p和miRNA-3075-3p。 TargetScan分析表明高迁移率组盒蛋白1 （HMGB1），AF4/FMR2家族成员3（AFF3）和EDIL3是miRNA-215-3p，miRNA-129-5p的潜在靶标和miRNA-3075。基于我们已发表的结果和令人兴奋的新初步数据，我们假设 H19诱导的Circrna-Edil3上调通过隔离在胆汁淤积性肝损伤中起关键作用 mirnas。提出了两个具体目标来检验我们的假设。目的1。检查H19诱导的作用在调节使用胆管细胞增殖，衰老和肝发炎的circrna-eDil3 胆固性肝损伤小鼠模型和人类PSC患者肝脏和血清样品。目标2：到确定H19诱导的Circrna-Edil3促进胆汁淤积性肝损伤的机制。我们将使用多种新建立的方法和动物模型来检验我们的假设。这些完成具体目的应阐明参与LNCRNA H19介导的潜在细胞/分子机制胆汁淤积性肝病的进展并确定新颖的诊断和治疗靶标。自效果以来胆汁淤积是深刻而广泛的，导致肝病恶化和全身性疾病恶化，该提案的主题是及时的，重要的，并且具有直接的临床应用。