Specific-sized hyaluronan: a dual targeted therapy for ALD

特定大小的透明质酸：ALD 的双重靶向治疗

基本信息

批准号：
9753072
负责人：
LAURA E. NAGY
金额：
$ 40.93万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9753072
关键词：
Address Affect Alcohol abuse Alcohol consumption Alcohol-Induced Disorders Alcoholic Hepatitis Alcoholic Liver Diseases American Animal Model Anti-inflammatory Biodistribution Bioinformatics Biological Assay Blood Circulation CD44 Antigens CD44 gene Caco-2 Cells Cells Chronic Cirrhosis Complex Data Development Devices Diet Disaccharides Disease Progression Elements Epithelial Epithelial Cells Ethanol Extracellular Matrix Fibrosis Funding Glucuronic Acids Goals HMMR gene Health Hepatic Hepatocyte Human Hyaluronan Hyaluronic Acid Immune Impairment Infiltration Inflammation Inflammation Mediators Inflammatory Inflammatory Response Injury Intestinal permeability Intestines Knock-out Kupffer Cells Leukocytes Lipopolysaccharides Liver Liver diseases Maintenance Mediating Medical MicroRNAs Morbidity - disease rate Mus Natural Immunity Nuclear Organ Organoids Pathogenesis Pathology Patients Peripheral Blood Mononuclear Cell Pilot Projects Polysaccharides Prevention strategy Process Production Publishing Rattus Receptor Signaling Regulatory Pathway Research Project Grants Rodent Model Signal Transduction TLR2 gene TLR4 gene Testing Therapeutic Therapeutic Agents Tight Junctions Translations United States National Institutes of Health Work alcohol exposure base beta-Defensins cell type clinical investigation cytokine effective therapy healthy volunteer human model insight intestinal epithelium liver development liver injury macrophage microbiome microbiota monocyte mortality mouse model next generation sequencing novel prevent problem drinker protective effect receptor recruit response rho stellate cell targeted treatment treatment strategy

项目摘要

ABSTRACT Alcoholic liver disease (ALD) develops in approximately 20% of alcoholics. The development of ALD is a complex process involving both parenchymal and non-parenchymal cells in the liver, as well as recruitment of immune cells in response to damage and inflammation. Innate immunity and inter-organ cross talk contribute to ethanol- induced liver injury with interactions between intestine and liver of particular importance. Impaired intestinal barrier function is associated with ethanol-induced liver injury in both humans and rodent models. Increased exposure of Kupffer cells, the resident hepatic macrophages, to gut-derived LPS during chronic ethanol activates TLR4-dependent production of inflammatory mediators. Chronic ethanol exposure also sensitizes Kupffer cells to LPS, resulting in increased production of inflammatory mediators. Hyaluronan (HA), an abundant extracellular matrix component, communicates with cells in a size-specific manner. Specific-sized HA fragments are either pro-inflammatory or anti-inflammatory, depending on the HA receptor and cell type involved in the response. We have discovered that specific-sized HA35 (average MW~35kD) normalizes TLR4- mediated signaling in Kupffer cells after chronic ethanol exposure and also protects mice from ethanol- induced gut and liver injury. By Next Generation Sequencing, we have identified a subset of microRNAs that are reciprocally regulated by HA35, providing novel insights into the mechanism of action for both ethanol and HA35 in regulating TLR4 signaling and macrophage polarization Furthermore, our team finds that specific-sized HA35 promotes intestinal health, at least in part by increasing expression of β-defensins and promoting the formation of tight junctions. Based on the multi-potent functions of HA35, here we will test the hypothesis that HA35 is a dually targeted therapeutic agent in ALD, normalizing Kupffer cell signal transduction after chronic ethanol exposure and protecting the intestinal epithelial barrier. We will address this hypothesis in three Specific Aims. Specific Aim 1: Investigate the mechanism for normalization of TLR4 signaling by HA35 in rat Kupffer cells and human PBMCs. Making use of both bioinformatics and cell based assays, we will 1) investigate the miRNA regulatory pathways reciprocally targeted by ethanol and HA35, which in turn regulate a) the control of nuclear-cytoplasmic shuttling and b) macrophage polarization. Specific Aim 2: Interrogate the impact of HA35 on maintenance of intestinal barrier function. We will use both cultured Caco-2 cells and mouse intestinal organoids to determine mechanisms for the direct effect of HA35 on protecting tight junctions from ethanol. Specific Aim 3: Test the ability of HA35 to prevent and treat chronic ethanol-induced intestinal and liver injury in mice. Importantly, medical grade HA for device-use is commercially available, thus enhancing the likelihood for a rapid translation of our studies on the dually protective functions of HA35 in chronic ethanol exposure into clinical investigations in ALD.

抽象的大约 20% 的酗酒者会患上酒精性肝病 (ALD)。 ALD 的发展是一个复杂的过程。涉及肝脏实质细胞和非实质细胞以及免疫细胞募集的过程对损伤和炎症的反应有助于乙醇- 肠道与肝脏之间的相互作用尤其重要，可引起肝损伤。在人类和啮齿动物模型中，屏障功能与乙醇引起的肝损伤有关。在长期乙醇摄入过程中，库普弗细胞（常驻肝巨噬细胞）暴露于肠道来源的脂多糖激活 TLR4 依赖性炎症介质的产生，慢性乙醇暴露也会导致过敏。枯否细胞转化为脂多糖，导致炎症介质（HA）的产生增加，透明质酸是一种丰富的物质。细胞外基质成分，以特定大小的方式与细胞通讯。片段是促炎的还是抗炎的，取决于HA受体和所涉及的细胞类型在响应中我们发现特定大小的HA35（平均MW~35kD）使TLR4-正常化。慢性乙醇暴露后库普弗细胞中介导的信号传导，还可以保护小鼠免受乙醇侵害通过下一代测序，我们已经鉴定出一部分 microRNA 受到 HA35 的相互调节，为乙醇和乙醇的作用机制提供了新的见解。 HA35 调节 TLR4 信号传导和巨噬细胞极化此外，我们的团队发现特定大小的 HA35 至少部分通过增加 β-防御素的表达和促进肠道健康来促进肠道健康基于HA35的多效功能，我们将检验以下假设： HA35 是 ALD 的双重靶向治疗剂，可使慢性酒精性肝病 (ALD) 后库普弗细胞信号转导正常化。乙醇暴露和保护肠上皮屏障我们将在三个具体方面讨论这一假设。具体目标 1：研究大鼠 Kupffer 中 HA35 使 TLR4 信号正常化的机制利用生物信息学和基于细胞的测定，我们将 1) 研究乙醇和 HA35 相互靶向的 miRNA 调控途径，反过来又调节 a) 的控制核-细胞质穿梭和 b) 巨噬细胞极化。具体目标 2：探讨 HA35 的影响。我们将使用培养的 Caco-2 细胞和小鼠肠道。类器官以确定 HA35 对保护紧密连接免受乙醇影响的直接作用机制。具体目标3：测试HA35预防和治疗慢性乙醇引起的肠和肝损伤的能力重要的是，用于设备用途的医用级 HA 已在市场上销售，从而增强了快速转化我们关于 HA35 在慢性乙醇中双重保护功能的研究的可能性参与 ALD 的临床研究。