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

• 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
• 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

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％的酗酒者中酒精性肝病（ALD）的发展。 ALD的发展是一个复杂的 过程涉及肝脏中的副群和非副作用细胞，以及免疫细胞的募集 响应损害和炎症。先天免疫和器官间交谈有助于乙醇 诱发肝损伤与肠道和肝之间的相互作用特别重要。肠道受损 屏障功能与人类和啮齿动物模型中乙醇诱导的肝损伤有关。增加 在慢性乙醇期间，kupffer细胞（居民肝巨噬细胞）暴露于肠道衍生的LP 激活炎症介质的TLR4依赖性产生。慢性乙醇暴露也感官 Kupffer细胞到LPS，导致炎症介质的产生增加。 Hyaluronan（HA），绝对 细胞外基质组件以大小特异性方式与细胞通信。特定尺寸的HA 片段是促炎或抗炎的，具体取决于涉及的HA接收器和细胞类型 在响应中。我们发现特异性HA35（平均MW〜35KD）使TLR4-归一化 慢性乙醇暴露后库普弗细胞中介导的信号传导，还保护小鼠免受乙醇的影响 诱发肠道和肝损伤。通过下一代测序，我们已经确定了一部分microRNA 受HA35的相互调节，为乙醇和乙醇的作用机理提供了新的见解 HA35在调节TLR4信号传导和巨噬细胞极化方面，我们的团队发现了特定尺寸的 HA35至少部分通过增加β-防御素的表达并促进肠道健康并促进肠道健康 紧密连接的形成。基于HA35的多功能功能，我们将在这里检验以下假设。 HA35是ALD中的双重靶向治疗剂，在慢性后标准化Kupffer细胞信号转导 乙醇暴露并保护肠上皮屏障。我们将以三个特定的特定来解决这一假设 目标。特定目标1：研究HA35在大鼠Kupffer中通过HA35归一化TLR4信号的机制 细胞和人类PBMC。利用生物信息学和基于细胞的测定法，我们将1）研究 miRNA调节途径是由乙醇和HA35靶向的，这又调节a）控制 核质洗车和b）巨噬细胞极化。特定目的2：询问HA35的影响 维持肠屏障功能。我们将同时使用培养的Caco-2细胞和小鼠肠道 器官确定HA35直接影响的机制对保护乙醇的紧密连接处的直接作用。 特定目标3：测试HA35预防和治疗慢性乙醇引起的肠损伤的能力 在老鼠中。重要的是，用于设备使用的医学级HA可商购，从而增强了 快速翻译我们对HA35在慢性乙醇中双重保护功能的研究的可能性 暴露于ALD中的临床研究。