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

基本信息

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
项目类别：
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

项目摘要

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，多项研究主要研究其修复、再生和免疫调节特性。使用不同的疾病动物模型表明，外源性 HLSC 治疗可改善包括肝脏疾病在内的急性器官损伤，其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。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) 及其相关的非编码 RNA (ncRNA) 知之甚少，它们可以介导促进进展和进展的遗传变化。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。在我们的初步研究中，大多数 ncRNA 的生物学功能尚不清楚。研究表明，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。通过令人信服的数据，我们提出了一个中心假设，即干细胞衍生的微泡中的 ncRNA 通过诱导肝胆组织的生长和抗衰老/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。细胞衍生的微泡并表征其组织修复的功能特性。在本应用中，我们提出系统评估干细胞衍生的 MV 中的干细胞依赖性 ncRNA 具有治疗胆汁淤积性肝损伤的潜力。假设重点关注以下具体目标：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性干性调节的 ncRNA 信号转导；其次，我们将识别参与存活和抗肿瘤的功能性 LPS/TGF-β 依赖性 miRNA。 -胆汁淤积性肝损伤期间的衰老第三，我们将确定富含干性相关的ncRNA的微泡对体内胆汁淤积性肝损伤的形态和功能恢复的影响。使用 BDL 或 CCl4 治疗以及在 Mdr2 缺陷小鼠中，来自具有抗 miRNA 或 T-UCR 过度表达的干细胞的 MV 对肝胆细胞生长、抗衰老和抗纤维化的治疗作用。所提出的研究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，所获得的关于干细胞衍生的 MV 的基本新知识有望推动总体进展。干细胞生物学领域。