Exosome platforms for assessment and therapy of chronic liver disease

用于评估和治疗慢性肝病的外泌体平台

• 批准号: 8968550
• 负责人: DAVID R BRIGSTOCK
• 金额: $ 21.49万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-10 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8968550
• 关键词: Accounting; Adult; Alcoholic Hepatitis; Alcoholic Liver Cirrhosis; Alcoholic Liver Diseases; Alcohols; American; Apoptosis; Applications Grants; Attenuated; Binding; Biogenesis; Biological; Biological Markers; Biology; Blood; Blood Circulation; Body Fluids; Cells; Cessation of life; Cicatrix; Cirrhosis; Collection; Complex; Complex Mixtures; Conditioned Culture Media; Data; Development; Diagnosis; Diagnostic; Disease; Distant; Economic Burden; Ethanol; Ethanol dependence; Fibrosis; Goals; Guidelines; Harvest; Health; Health Status; Hepatic Stellate Cell; Hepatocyte; Home environment; Homing; Image; In Vitro; Inflammation; Integrins; Knowledge; Lead; Ligands; Liver; Liver Fibrosis; Liver diseases; Mediating; Membrane; Messenger RNA; Methods; MicroRNAs; Molecular; Molecular Profiling; Multivesicular Body; Mus; Outcome; Pathway interactions; Patients; Population; Primary carcinoma of the liver cells; Production; Proteins; Reading; Reporting; Risk; Role; Sepsis; Serum; Severities; Site; Small Interfering RNA; Specimen; Stimulus; Testing; Therapeutic; Therapeutic Effect; Tissues; Treatment Efficacy; Urine; Vesicle; Work; alcohol effect; base; cell type; chronic liver disease; clay; connective tissue growth factor; differential expression; disease diagnosis; extracellular; health economics; improved; in vivo; in vivo Model; innovation; liver biopsy; liver injury; minimally invasive; mortality; nanoscale; nanosized; nanovesicle; non-alcoholic; novel; public health relevance; response; tool

 DESCRIPTION (provided by applicant): The broad long-term objective is to improve methods of disease assessment and treatment in the liver. Approximately 5.5 million American adults (i.e., 2-3% of the adult US population) suffer from chronic liver disease. Alcoholic liver disease (ALD) accounts for the largest proportion of chronic liver disease in the West and exacerbates liver injury due to non-alcoholic causes. Alcoholic cirrhosis ranks as the 8th most common cause of mortality in the US and the second leading cause of GI-related deaths. There is an unmet need to improve our arsenal of diagnostic options by using modern molecular approaches to analyze biological specimens that can be collected in a minimally-invasive or non-invasive manner, and a new lead has emerged with the discovery of "exosomes". These are nanovesicles that are secreted by many cell types and which contain a complex mixture of microRNAs, mRNAs and proteins that reflect the transcriptional and/or translational activity of their producer cells. We showed that (i) miRs in circulating exosomes undergo dynamic changes during experimental fibrosis in mice; (ii) expression of fibrosis-related miRs or mRNA in exosomes secreted by hepatic stellate cells (HSC, the principal pro-fibrogenic cell type in the liver) reflects the induction status of fibrogenic pathways within the cells; (iii) expression in HC of molecules regulating exosome biogenesis is ethanol-dependent; (iv) fibrosis-related miRs are exosomally shuttled between HSC and are functional after being taken up by recipient cells; and (v) exosomes from HSC are preferentially taken up by HSC both in vivo and in vitro, highlighting the existence of a novel homing mechanism for HSC-derived exosomes. Thus our overall objective is to establish the role of exosomes in liver disease and their potential role in disease diagnosis or therapy. Our central hypothesis is that exosomes have diagnostic and therapeutic utility in liver disease, including that caused by alcohol. The Aims to test our hypothesis are: Specific Aim 1: Establish ethanol-mediated changes in exosome production and molecular payload. We will determine the effect of ethanol on pathways of exosome biogenesis and secretion in HSC and establish miR signatures in exosomes from the serum of ALD patients Specific Aim 2. Establish mechanisms and therapeutic applications of exosome homing to HSC. We will characterize integrin-mediated interactions between exosomes and HSC and, in an in vivo model of experimental liver fibrosis, establish their role in mediating HSC-specific localization and therapeutic efficacy of exosomes loaded with anti-fibrotic siCTGF. The expected outcome of these exploratory R21 studies will be innovative breakthroughs in the diagnosis and therapy of chronic liver disease by improved understanding and exploitation of exosome function. The rationale underlying the application is that identification of exosomal molecular payloads and intercellular shuttling mechanisms will have direct relevance to disease diagnosis and therapy. The positive impact of these studies is that they will improve the health of millions of people world-wide with ALD and other liver diseases.

 描述（由申请人提供）：广泛的长期目标是改善肝脏疾病评估和治疗方法。大约550万美国成年人（即，2-3%的美国成年人患有慢性肝病。酒精性肝病（ALD）在西方慢性肝病中占最大比例，并且由于非酒精性原因而加剧肝损伤。酒精性肝硬化是美国第八大最常见的死亡原因，也是GI相关死亡的第二大原因。通过使用现代分子方法来分析可以以微创或非侵入性方式收集的生物标本来改进我们的诊断选项库存在未满足的需求，并且随着“外来体”的发现而出现了新的线索。这些是由许多细胞类型分泌的纳米囊泡，并且其含有反映其生产细胞的转录和/或翻译活性的微小RNA、mRNA和蛋白质的复杂混合物。我们发现（i）在小鼠实验性纤维化过程中，循环外泌体中的miR经历动态变化;（ii）肝星状细胞分泌的外泌体中纤维化相关miR或mRNA的表达，（iii）调节外泌体生物发生的分子在HC中的表达是乙醇依赖性的;（iv）纤维化相关的miR在HSC之间外泌体穿梭，并且在被受体细胞摄取后是功能性的;和（v）来自HSC的外泌体在体内和体外都优先被HSC摄取，突出了HSC衍生的外泌体的新归巢机制的存在。因此，我们的总体目标是确定外来体在肝脏疾病中的作用及其在疾病中的潜在作用。 诊断或治疗。我们的中心假设是外泌体在肝病中具有诊断和治疗作用，包括酒精引起的肝病。测试我们假设的目的是：具体目的1：建立乙醇介导的外泌体产生和分子有效载荷的变化。我们将确定乙醇对HSC中外来体生物发生和分泌途径的影响，并在来自ALD患者血清的外来体中建立miR特征。建立外泌体归巢HSC的机制和治疗应用。我们将表征整合素介导的外泌体和HSC之间的相互作用，并在实验性肝纤维化的体内模型中，建立它们在介导HSC特异性定位和加载有抗纤维化siCTGF的外泌体的治疗功效中的作用。这些探索性R21研究的预期结果将是通过改善对外泌体功能的理解和利用，在慢性肝病的诊断和治疗方面取得创新突破。该申请的基本原理是，外泌体分子有效载荷和细胞间穿梭机制的鉴定将与疾病诊断和治疗直接相关。这些研究的积极影响是，它们将改善全球数百万ALD和其他肝病患者的健康状况。