Development of delivery methods for combination microRNA treatment of alcohol-associated liver disease

开发联合 microRNA 治疗酒精相关性肝病的递送方法

• 批准号: 10207371
• 负责人: David Oupicky
• 金额: $ 38.9万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207371
• 关键词: Address; Affect; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Animal Model; Attenuated; Aziridines; Biological; CXCR4 Receptors; CXCR4 gene; Cholesterol; Chronic; Cirrhosis; Clinical Trials; Collagen; Combined Modality Therapy; Complex; Data; Development; Disease; Down-Regulation; Drug Delivery Systems; Drug Kinetics; Effectiveness; Encapsulated; Event; Exhibits; Exposure to; Fatty Liver; Fibrosis; Formulation; Functional disorder; Genes; Goals; Health; Hepatic; Hepatic Stellate Cell; Hepatocyte; Human; In Vitro; Inflammatory Response; Knowledge; Kupffer Cells; Ligands; Ligation; Liver; Liver Cirrhosis; Liver Fibrosis; Liver diseases; Mannose; Mediating; Methods; MicroRNAs; Microfluidics; Modeling; Modification; Molecular Weight; Mus; Natural History; Pathogenesis; Pathway interactions; Performance; Pharmacology; Play; Polymers; Positioning Attribute; Production; Property; Public Health; Research; Role; Signal Transduction; Slice; Stromal Cell-Derived Factor 1; TNF gene; Testing; Therapeutic; Therapeutic Effect; Transfection; Treatment Efficacy; Tropism; Work; alcohol abuse therapy; antifibrotic treatment; attenuation; base; bile duct; chemokine receptor; design; immunoreaction; improved; in vivo evaluation; inhibitor/antagonist; innovation; intrahepatic; liver injury; macrophage; mouse model; nanoparticle; nanoparticle delivery; novel therapeutic intervention; overexpression; particle; prototype; targeted delivery; therapeutic evaluation; therapeutic miRNA; therapeutic target

PROJECT SUMMARY Alcohol-associated liver disease (AALD) is a major and growing health concern with limited treatment options. The natural history of AALD (formerly known as alcoholic liver disease) includes fatty liver disease, alcoholic hepatitis and the development of fibrosis preceding end-stage cirrhosis. MicroRNAs (miRNAs) represent a new class of therapeutics due to their ability to simultaneously affect multiple fibrosis-associated pathways. Among possible targets, miR-155 is involved in inflammatory responses mediated by Kupffer cells (KCs) that affect fibrogenic events in multiple other hepatic cells. Chemokine receptor CXCR4 and its cognate ligand stromal cell- derived factor-1 play important and complex roles in the pathogenesis of AALD, including the coordination of the initial immune reaction upon liver injury and later in controlling the progression of liver fibrosis through its activating effect on hepatic stellate cells (HSCs) and collagen production. The goal of this project is to develop integrated miRNA delivery platform based on self-assembled nanoparticles (polyplexes) that deliver anti-miR- 155 to activated KCs and in parallel inhibit CXCR4 signaling in activated HSCs in the liver. The delivery platform is based on innovative CXCR4 inhibitors based on cyclam-modified low molecular weight poly(ethylenimine)s (C-PEI) that efficiently encapsulate and systemically deliver miRNA. The objective is to test the hypothesis that C-PEI/miRNA will lead to enhanced combination effect due to attenuation of profibrogenic signaling of both hepatic macrophages and the matrix-producing HSCs. We will accomplish the overall objectives in three specific aims. In Aim 1, we will optimize formulation of the C-PEI-miRNA nanoparticles that deliver anti-miR-155 in liver fibrosis. Based on encouraging antifibrotic activity in our preliminary studies, we hypothesize that polyplex modification with mannose and with stabilizing cholesterol moieties will result in efficient delivery of the miRNA to activated KCs, while excess free C-PEI will target activated HSCs. In Aim 2, we will test the in vivo therapeutic efficacy of the polyplexes in the bile-duct ligation (BDL) model and mouse models of chronic alcohol administration with exposure to CCl4. The goal is to conduct comprehensive evaluation of therapeutic efficacy of the optimized C-PEI-Chol/anti-miR-155 polyplexes in different models and multiple stages of fibrosis. The findings from the mouse models will be validated in cultured human precision-cut liver slices. In Aim 3, we will determine the mechanism of antifibrotic activity of the combined miR-155 and CXCR4 inhibition. Although our preliminary data are consistent with miR-155 downregulation in KCs and inhibition of CXCR4 in activated HSCs, the precise mechanism of action of the C-PEI/anti-miR-155 polyplexes is not known. Therefore, the studies in this aim will be designed to ascertain the underlying mechanisms of action of the polyplexes. Overall, this project will contribute to the fundamental understanding of the role and therapeutic potential of CXCR4 and miR-155 inhibition in AALD and will innovatively address targeted delivery of drug/miRNA combinations as an anti-fibrotic treatment.

项目摘要 与酒精相关的肝病（AALD）是有限的治疗选择，这是一个主要且日益严重的健康问题。 AALD的自然历史（以前称为酒精性肝病）包括脂肪肝疾病，酒精性 肝炎和终末期肝硬化之前的纤维化发展。 microRNA（miRNA）代表一个新的 由于它们具有同时影响多个纤维化相关途径的能力，因此治疗剂类别。之中 可能的靶标，miR-155参与了影响库普弗细胞（KC）介导的炎症反应 其他多个肝细胞中的纤维化事件。趋化因子受体CXCR4及其同源配体基质细胞 - 衍生因子1在AALD的发病机理中起重要而复杂的作用，包括协调 肝损伤时的初始免疫反应，后来控制肝纤维化的进展 激活对肝星状细胞（HSC）和胶原蛋白产生的作用。该项目的目的是开发 基于自组装的纳米颗粒（多流线）的集成miRNA递送平台，该平台提供抗MiR- 155以激活的KC和平行抑制肝脏活化HSC中的CXCR4信号传导。交付平台 基于创新的CXCR4抑制剂，基于圆柱体修饰的低分子量聚（乙烯亚胺）S （C-PEI）有效地封装并系统地传递miRNA。目的是检验以下假设 C-PEI/miRNA将由于两者的衰减信号的衰减而导致组合效应增强 肝巨噬细胞和产生基质的HSC。我们将以三个特定的特定目标实现总体目标 目标。在AIM 1中，我们将优化c-PEI-MIRNA纳米颗粒的配方 纤维化。基于鼓励我们的初步研究的抗纤维化活性，我们假设息肉 用甘露糖和稳定胆固醇部分进行修饰将导致miRNA有效递送 激活的KC，而多余的自由C-PEI将靶向活化的HSC。在AIM 2中，我们将测试体内治疗 多流线在胆道连接（BDL）模型和慢性酒精的小鼠模型中的功效 暴露于CCL4的管理。目的是对治疗功效进行全面评估 在不同模型和多个纤维化阶段中，优化的C-PEI-CHOL/ANTI-MIR-155多型多路复管。这 小鼠模型的发现将在培养的人类精确切割肝切片中进行验证。在AIM 3中，我们将 确定联合miR-155和CXCR4抑制的抗纤维化活性的机制。虽然我们的 初步数据与KC中的miR-155下调和激活的HSC中CXCR4的抑制一致， C-PEI/Anti-MIR-155多链体的确切作用机理尚不清楚。因此，研究 该目标将旨在确定多体链体的基本作用机理。总体而言，这个项目 将有助于对CXCR4和MIR-155的作用和治疗潜力的基本了解 AALD中的抑制作用，并将创新地解决针对药物/miRNA组合作为抗纤维化的递送 治疗。