LncRNA H19 in Cholestatic Liver Diseases

LncRNA H19 在胆汁淤积性肝病中的作用

• 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
• 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

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）是一种发病率和死亡率都很高的胆汁淤积性肝病。 由于胆汁形成或流动受损导致的胆管炎症和纤维化损伤代表了主要的 PSC的特点。大多数哺乳动物基因组是非蛋白质编码序列。小于 人类基因组的2%由蛋白质编码基因组成。很大一部分转录的哺乳动物 基因组是非编码RNA（ncRNA），包括长非编码RNA（lncRNA）、环状RNA（circRNA）、 和微小RNA（miRNA），但其中只有少数进行了结构注释。的生物学功能 ncRNA在很大程度上仍是未知的。高通量RNA测序（RNAseq）和circRNA- 特定的生物信息学算法已经鉴定了数千种具有组织特异性表达模式的circRNA。 然而，circRNA在疾病中的相关性和功能，特别是在肝脏疾病中，仍有待进一步研究。 测定我们以前的研究报告，结合初级胆汁酸（CBA），牛磺胆酸（TCA）， 激活1-磷酸鞘氨醇受体2（S1 PR 2）并诱导胆管细胞H19表达。TCA- 诱导的H19上调是通过激活炎性细胞因子而导致胆汁淤积性肝损伤的主要促进因素 巨噬细胞和肝星状细胞（HSC）。我们使用Arraystar circRNAseq的新初步数据确定了 33种重叠circRNA，在Mdr 2-/-小鼠中显著上调，在Mdr 2-/-/H19-小鼠中下调。 /-小鼠。其中，mmu_circRNA_26644（circRNA-Edil 3），其由表皮生长因子样因子编码。 重复序列和盘状结构域3（Edil 3）基因，是Mdr 2-/- /H19-/-小鼠。序列分析表明，circRNA-Edil 3可能充当miRNA-215- 3 p的“海绵”， miRNA-129-5p和miRNA-3075-3p。TargetScan分析表明，高迁移率族蛋白1 HMGB 1、AF 4/FMR 2家族成员3（Aff 3）和Edil 3是miRNA-215- 3 p、miRNA-129- 5 p的潜在靶点 和miRNA-3075。根据我们发表的结果和令人兴奋的新的初步数据，我们假设， H19诱导的circRNA-Edil 3上调通过螯合在胆汁淤积性肝损伤中起关键作用 miRNAs。提出了两个具体的目标来检验我们的假设。目标1.研究H19诱导的 使用circRNA-Edi 1 3调节胆管细胞增殖、衰老和肝脏炎症 胆汁淤积性肝损伤小鼠模型和人PSC患者肝脏和血清样品。目标2： 鉴定H19诱导的circRNA-Edi 13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验证我们的假设。完成这些 具体的目的应该阐明潜在的细胞/分子机制参与lncRNA H19介导的 胆汁淤积性肝病的进展，并确定新的诊断和治疗靶点。由于影响 胆汁淤积的影响是深刻和广泛的，导致肝脏疾病和全身性疾病恶化， 该提案的主题是及时的，重要的，并具有直接的临床应用。