Hepatocyte-derived extracellular vesicles in alcoholic liver disease

酒精性肝病中肝细胞源性细胞外囊泡

基本信息

批准号：
10381729
负责人：
Ariel Feldstein
金额：
$ 58.13万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10381729
关键词：
Address Alcoholic Fatty Liver Alcoholic Hepatitis Alcoholic Liver Diseases Alcoholic steatohepatitis Animal Model Bar Codes Biological Markers Biology Blood Blood Circulation Cell Communication Cells Cellular biology Chronic Country DNA analysis DNA biosynthesis Data Deoxyribonucleotides Development Diagnosis Disease Disease Progression Effector Cell Encapsulated Enzymes Fatty Liver Fibrosis Generations Genes Hepatic Hepatic Stellate Cell Hepatocyte Human Infiltration Inflammasome Inflammation Inflammatory Interleukin-17 Investigation Lipids Liquid substance Liver Liver Fibrosis Macrophage Activation Mediating Messenger RNA MicroRNAs Mitochondrial DNA Modeling Molecular Morbidity - disease rate Mus Organelles Oxides Pathologic Pathology Pathway Analysis Pathway interactions Patients Pattern Penetration Phenotype Plasma Ploidies Process Proteins RNA Role Severity of illness Signal Pathway Signal Transduction Small Interfering RNA Staging Sterility Stress Surface Tail Testing Therapeutic Toll-like receptors Translations United States Untranslated RNA Veins base chronic alcohol ingestion chronic liver disease design efficacious treatment exosome extracellular extracellular vesicles in vitro Assay in vivo liver biopsy macrophage mortality mouse model nonalcoholic steatohepatitis noninvasive diagnosis novel prevent response therapeutic RNA transcriptome sequencing transcriptomics vesicular release

项目摘要

Alcoholic liver disease (ALD) continues to be a devastating disease in the United States and many other countries with the global mortality estimated to exceed 0.7 million per year. Chronic ethanol consumption results in lipid accumulation in hepatocytes and their organelle stress leading to inflammation and fibrosis in the progression of ALD. The crosstalk between hepatocytes and non- parenchymal cells - including hepatic macrophages (HMs) and hepatic stellate cells (HSCs) - is crucial to this process. However, the molecular mechanisms and signaling pathways involved in the crosstalk between lipid overloaded hepatocytes and non-parenchymal cells, remain incompletely understood. In particular, the mechanisms by which this crosstalk facilitates or regulates a transition from chronic asymptomatic alcoholic steatohepatitis (ASH) to fatal alcoholic hepatitis (AH) is a central issue for designing efficacious novel treatments for this devastating disease. While focused on the investigation of cell-to-cell communication we recently revealed that damaged hepatocytes release extracellular vesicles (EVs) and these EVs circulate in the blood in mouse models of ALD. EVs are efficiently internalized into target cells and transfer their cargo including miRNAs. The later is a key mechanism by which encapsulated miRNAs in EVs (EV-miRNAs) serve as “functional extracellular RNAs” to regulate protein translation in target cells. EVs also contain sterile danger signal known as damage-associated molecular patterns (DAMPs). We found that EVs derived from hepatocytes from AH mice are enriched in mitochondrial DNA (mtDNA) that contribute to activation of an inflammatory process. Further, our pathway analysis of RNA-seq data comparing AH vs. preceding chronic (c)ASH mouse livers, identifies EV as one of the most significantly upregulated pathways. Based on these results, we propose the central hypothesis that EVs with a quantitatively and qualitatively distinct cargo are released from damaged hepatocytes in AH vs. preceding cASH to induce unique cellular crosstalk in the genesis of the AH pathologic phenotype. We also hypothesize that plasma EV analysis serves as a liquid liver biopsy providing a barcode for diagnosis and staging of ALD severity. To investigate these hypotheses our proposal has the following SPECIFIC AIMS. 1) Determine the role of EVs and their cargo derived from hepatocytes as barcodes for a transition from asymptomatic cASH to AH in murine models and human ALD. 2) Dissect the mechanisms involved in EV-mediated cell-to-cell communication in the cASH to AH transition. To address these central issues, we have put together a MPI investigative team with expertise in EV biology, ALD pathology, development of unique animal models, human ALD, and RNA therapeutics.

酒精性肝病（ALD）在美国和许多国家仍然是一种毁灭性的疾病全球每年因慢性乙醇死亡的人数估计超过 70 万人。消耗导致肝细胞中的脂质积累及其细胞器应激，导致 ALD 进展中的炎症和纤维化。肝细胞和非肝细胞之间的串扰。实质细胞 - 包括肝巨噬细胞 (HM) 和肝星状细胞 (HSC) - 是然而，参与这一过程的分子机制和信号通路至关重要。脂质超载的肝细胞和非实质细胞之间的串扰仍然存在特别是，这种串扰促进或促进的机制尚不完全清楚。调节从慢性无症状酒精性脂肪性肝炎 (ASH) 到致命性酒精性肝炎的转变肝炎（AH）是针对这种毁灭性的肝炎设计有效的新型治疗方法的一个核心问题在专注于细胞间通讯的研究时，我们最近揭示了这一点。受损的肝细胞释放细胞外囊泡（EV），并且这些 EV 在在 ALD 小鼠模型中，血液被有效地内化到靶细胞中并转移它们。后者是将 miRNA 封装在 EV 中的关键机制。 (EV-miRNA) 作为“功能性细胞外 RNA”来调节靶标中的蛋白质翻译 EV 细胞还包含无菌危险信号，称为损伤相关分子模式。 (DAMP)。我们发现来自 AH 小鼠肝细胞的 EV 富含线粒体 DNA (mtDNA) 有助于激活炎症过程。比较 AH 与先前慢性 (c)ASH 小鼠肝脏的 RNA-seq 数据的通路分析，根据这些结果，我们将 EV 确定为最显着上调的途径之一。中心假设是，具有数量和质量不同的货物的电动汽车 AH 中受损肝细胞与之前的 cASH 中的释放物相比，诱导独特的细胞串扰我们还进行了血浆 EV 分析。作为液体肝活检，为 ALD 严重程度的诊断和分期提供条形码。调查这些假设，我们的提案有以下具体目标 1) 确定。 EV 及其源自肝细胞的货物作为条形码的作用，用于从小鼠模型和人类 ALD 中无症状 cASH 至 AH 2) 剖析机制。参与 cASH 到 AH 转变过程中 EV 介导的细胞间通讯。这些核心问题，我们组建了一个具有 EV 生物学专业知识的 MPI 调查团队， ALD 病理学、独特动物模型的开发、人类 ALD 和 RNA 疗法。