The Role of Stem Cell Derived Microvesicles in Cholestatic Liver Injury

干细胞衍生的微泡在胆汁淤积性肝损伤中的作用

• 批准号: 9930828
• 负责人: Gianfranco D Alpini
• 金额: $ 40.22万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9930828
• 关键词: Acute; Address; Alcoholic Liver Diseases; Animal Disease Models; Animal Model; Applications Grants; Attention; Biliary; Bioinformatics; Biological; Biological Assay; Biological Process; Cell Aging; Cell Line; Cell Proliferation; Cell physiology; Cells; Characteristics; Chronic; Chronic Disease; Data; Development; Disease; Epithelial Cells; Evaluation; Extrahepatic Bile Ducts; Family member; Fibrosis; Flow Cytometry; Gene Expression; Gene Transfer; Genes; Genetic; Growth; Health; Hepatic; Hepatobiliary; Hepatocyte; Human; In Vitro; Inflammatory; Inflammatory Response; Injury; Knowledge; Lead; Life; Ligation; Lipopolysaccharides; Liver; Liver Stem Cell; Liver diseases; Mediating; Mesenchymal; Mesenchymal Stem Cells; Messenger RNA; MicroRNAs; Microarray Analysis; Morphology; Mus; Mutation; Natural regeneration; Organ; Phenotype; Primary biliary cirrhosis; Property; RNA; Recovery; Recovery of Function; Regulation; Research; Rest; Ribonucleases; Role; SCID Mice; Self Tolerance; Signal Transduction; Stem cells; TLR4 gene; Techniques; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Transcriptional Regulation; Transforming Growth Factor beta; Untranslated RNA; Western Blotting; acute liver disease; acute liver injury; base; bile duct; cell growth; cholangiocyte; cholestatic liver disease; cytokine; effective therapy; gene product; immunoregulation; in vivo; intrahepatic; knock-down; liver injury; microvesicles; mouse model; novel; novel therapeutic intervention; overexpression; paracrine; pre-clinical; primary sclerosing cholangitis; programs; public health relevance; reconstruction; regenerative; response; senescence; stem cell biology; stem cell therapy; stemness; tissue repair

 DESCRIPTION (provided by applicant): The functions of the human liver are essential to life since the liver is the only organ capable of regeneration. Human liver stem cells (HLSCs) have been extensively studied for their reparative, regenerative and immunomodulatory properties. Several studies, using different animal models of diseases, showed that treatment with exogenous HLSCs ameliorates acute organ injury including hepatic disorders. The mechanisms may involve paracrine factors promoting proliferation of surviving intrinsic epithelial cells. Whil stem cell therapies have been in pre-clinical use for the treatment of liver diseases, very little s known about the stem cell derived microvesicles (MVs) and their related non-coding RNAs (ncRNAs), which can mediate genetic changes that promote the progression and recovery of liver disorders. Many ncRNAs are expressed in a tissue-specific manner that is aberrantly altered in human alcoholic liver injuries. The biological function of the majority of ncRNAs in livr diseases is undefined. In our preliminary studies, we have shown that selected ncRNA genes are altered after cholestatic liver injuries, and aberrantly expressed in human liver stem cells and their derived MVs that can modulate the response to liver injury as well as cell anti-senescence and remodeling potentials. Based on these compelling data, we propose the central hypothesis that ncRNAs in stem cell derived microvesicles contribute to the recovery of cholestatic liver injury through induction of growth and anti-senescence/anti-fibrosis of hepatobiliary tissues and cells. To test this hypothesis, we have established techniques for ncRNA gene manipulation, functional and interaction analysis, and generated stably transfected or knockdown cell lines as well as animal models of cholestatic liver injury. Our long-term objective is to identify and isolate stem cell derived microvesicles and to characterize their functional properties of tissue repair. In this application, we propose the systematic evaluation o stemness dependent ncRNAs as markers in stem cell derived MVs with the therapeutic potentials for cholestatic liver injury. We will address our central hypothesis by focusing on the following specific aims: First, we will define the functional stemness regulated ncRNAs signaling involved in tissue repair-related cellular functions in hepatobiliary cells. Second, we will identiy functional LPS/TGF-ß dependent miRNAs involved in survival and anti-senescence during cholestatic liver injury. Third, we will determine the effects of stemness related ncRNAs enriched microvesicles on accelerating the morphologic and functional recovery of cholestatic liver injury in vivo using BDL or CCl4 treatment and in Mdr2 deficient mice, the genetic mouse model of PSC. Therapeutic effects of MVs derived from stem cells with anti-miRNAs or over-expression of T-UCRs on hepatobiliary cell growth, anti- senescence and anti-fibrosis will be evaluated. The results of the proposed studies may lead to new therapeutic strategies for human cholestatic liver diseases. Meanwhile, the acquired fundamental new knowledge about stem cell derived MVs is expected to advance the general field of stem cell biology.

 描述（由适用提供）：人肝的功能对生命至关重要，因为肝脏是唯一能够再生的器官。人肝干细胞（HLSC）已因其修复，再生和免疫调节特性而被广泛研究。使用不同疾病动物模型的几项研究表明，外源HLSCS治疗可以改善包括肝疾病在内的急性器官损伤。该机制可能涉及旁分泌因子，促进固有上皮细胞的增殖。尽管干细胞疗法一直用于治疗肝病的治疗，但对干细胞衍生的微囊泡（MVS）及其相关的非编码RNA（NCRNA）的了解鲜为人知，这可以介导遗传变化，从而促进肝脏疾病的进展和恢复。许多NCRNA以组织特异性的方式表达，在人类酒精肝损伤中异常改变。 LIVR疾病中大多数NCRNA的生物学功能是不确定的。在我们的初步研究中，我们表明，胆汁淤积性肝损伤后所选的NCRNA基因会改变，并在人肝干细胞中异常表达及其衍生的MV，可以调节对肝损伤的反应以及细胞抗测定和重塑潜力。基于这些引人注目的数据，我们提出了一个中心假设，即干细胞中的NCRNA通过诱导生长和肝胆管组织和细胞的抗味/抗纤维化/抗纤维化，有助于恢复胆固性肝损伤。为了检验这一假设，我们建立了用于NCRNA基因操纵，功能和相互作用分析的技术，并产生了稳定的翻译或敲低细胞系以及胆固醇肝损伤的动物模型。我们的长期目标是识别和分离干细胞得出的微泡并表征其组织修复的功能特性。在此应用中，我们提出了系统评估O干性依赖性NCRNA作为具有胆固醇肝损伤的治疗潜力的干细胞中的标记。我们将通过关注以下特定目的来解决我们的中心假设：首先，我们将定义与肝胆管细胞中组织修复相关细胞功能涉及的功能性干性NCRNA信号传导。其次，我们将在胆固醇肝损伤过程中确定与生存和抗衰变有关的功能性LPS/TGF-β。第三，我们将使用BDL或CCL4处理以及MDR2缺陷小鼠（PSC的遗传小鼠模型）确定富含微囊泡富含微泡的NCRNA对加速度胆固性肝损伤的形态和功能恢复的影响。将评估源自具有抗MIRNA的干细胞的MVS的治疗作用或T-UCRS对肝胆管细胞生长，抗味和抗纤维化的过表达。拟议研究的结果可能会导致针对人类胆汁肝疾病的新治疗策略。同时，获得的有关干细胞得出的MVS的基本知识预计将推进干细胞生物学的一般领域。