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: 10227144
• 负责人: LAURA E. NAGY
• 金额: $ 41.19万
• 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/10227144
• 关键词: Address; Affect; Alcohol abuse; Alcohol consumption; Alcohol-Induced Disorders; Alcoholic Hepatitis; Alcoholic Liver Diseases; American; Animal Model; Anti-Inflammatory Agents; Biodistribution; Bioinformatics; Biological Assay; Blood Circulation; CD44 Antigens; CD44 gene; Caco-2 Cells; Cells; Chronic; Cirrhosis; Complex; Data; Development; Devices; Disaccharides; Disease Progression; Elements; 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; 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; dietary; effective therapy; healthy volunteer; hepatocellular injury; human model; insight; intestinal barrier; intestinal epithelium; intestinal injury; 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 以应对损伤和炎症。先天免疫和器官间串扰有助于乙醇- 肠道和肝脏之间的相互作用特别重要，可引起肝损伤。肠道受损 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。增加 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。长期接触乙醇也会导致过敏 库普弗细胞转化为脂多糖，导致炎症介质的产生增加。透明质酸 (HA) 是一种丰富的 细胞外基质成分，以特定大小的方式与细胞通讯。特定尺寸的HA 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 在回应中。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 诱发肠道和肝脏损伤。通过下一代测序，我们已经鉴定出 microRNA 的一个子集 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 形成紧密连接。基于HA35的多能功能，这里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。我们将在三个具体的方面来解决这个假设 目标。具体目标 1：研究大鼠 Kupffer 中 HA35 使 TLR4 信号传导正常化的机制 细胞和人类 PBMC。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：质疑 HA35 的影响 维持肠道屏障功能。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 在小鼠中。重要的是，用于设备用途的医疗级 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。