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（申请人提供）：人类肝脏的功能对于生命至关重要，因为肝脏是唯一能够再生的器官。人类肝脏干细胞（HLSC）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，外源性 HLSC 治疗可改善急性器官损伤，包括肝脏疾病。该机制可能涉及促进存活的内在上皮细胞增殖的旁分泌因子。虽然干细胞疗法已在临床前用于治疗肝脏疾病，但人们对干细胞衍生的微泡 (MV) 及其相关非编码 RNA (ncRNA) 知之甚少，它们可以介导促进肝脏疾病进展和恢复的遗传变化。许多 ncRNA 以组织特异性方式表达，这种方式在人类酒精性肝损伤中发生异常改变。大多数 ncRNA 在肝病中的生物学功能尚不清楚。在我们的初步研究中，我们已经表明，选定的 ncRNA 基因在胆汁淤积性肝损伤后发生改变，并在人肝干细胞及其衍生的 MV 中异常表达，可以调节对肝损伤的反应以及细胞抗衰老和重塑潜力。基于这些令人信服的数据，我们提出了一个中心假设：干细胞衍生的微泡中的 ncRNA 通过诱导肝胆组织和细胞的生长和抗衰老/抗纤维化，有助于胆汁淤积性肝损伤的恢复。为了检验这一假设，我们建立了 ncRNA 基因操作、功能和相互作用分析技术，并生成了稳定转染或敲低的细胞系以及胆汁淤积性肝损伤的动物模型。我们的长期目标是识别和分离干细胞衍生的微泡，并表征其组织修复的功能特性。在此应用中，我们建议系统评估干性依赖性 ncRNA 作为干细胞衍生 MV 中的标记物，具有治疗胆汁淤积性肝损伤的潜力。我们将通过关注以下具体目标来解决我们的中心假设：首先，我们将定义参与肝胆细胞组织修复相关细胞功能的功能干性调节的 ncRNA 信号传导。其次，我们将鉴定在胆汁淤积性肝损伤期间参与存活和抗衰老的功能性 LPS/TGF-β 依赖性 miRNA。第三，我们将确定干性相关的 ncRNA 富集微泡对使用 BDL 或 CCl4 治疗体内加速胆汁淤积性肝损伤的形态和功能恢复以及在 Mdr2 缺陷小鼠（PSC 遗传小鼠模型）中的影响。将评估源自具有抗 miRNA 或 T-UCR 过表达的干细胞的 MV 对肝胆细胞生长、抗衰老和抗纤维化的治疗效果。拟议研究的结果可能会为人类胆汁淤积性肝病带来新的治疗策略。同时，所获得的有关干细胞衍生MV的基础新知识有望推动干细胞生物学的一般领域的发展。