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: 10676945
• 负责人: BENITA L. MCVICKER
• 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/10676945
• 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; 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; Macrophage; Mannose; Mediating; Methods; MicroRNAs; Microfluidics; Modeling; Modification; Molecular Weight; Mus; Natural History; Pathogenesis; Pathway interactions; Performance; 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; bile duct; chemokine receptor; design; fatty liver disease; immunoreaction; improved; in vivo evaluation; inhibitor; innovation; intrahepatic; liver injury; microRNA delivery; mouse model; nanoparticle; nanoparticle delivery; novel therapeutic intervention; overexpression; particle; pharmacologic; prototype; self assembly; 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.

项目概要 酒精相关性肝病 (AALD) 是一个日益严重的主要健康问题，但治疗选择有限。 AALD（以前称为酒精性肝病）的自然史包括脂肪肝病、酒精性肝病 肝炎和终末期肝硬化之前纤维化的发展。 MicroRNA（miRNA）代表了一种新的 由于它们能够同时影响多种纤维化相关途径，因此被称为一类治疗方法。之中 miR-155 参与库普弗细胞 (KC) 介导的炎症反应，影响 多种其他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 的发病机制中发挥着重要而复杂的作用，包括协调 肝损伤时的初始免疫反应，以及随后通过其控制肝纤维化的进展 对肝星状细胞（HSC）和胶原蛋白产生的激活作用。该项目的目标是开发 基于自组装纳米粒子（多聚体）的集成 miRNA 递送平台，可递送抗 miR- 155 作用于激活的 KC，同时抑制肝脏中激活的 HSC 中的 CXCR4 信号传导。配送平台 基于创新的 CXCR4 抑制剂，该抑制剂基于 Cycam 改性的低分子量聚（乙烯亚胺） （C-PEI）可有效封装并系统地递送 miRNA。目的是检验以下假设： 由于两者的促纤维化信号减弱，C-PEI/miRNA 将导致增强的组合效应 肝巨噬细胞和产生基质的 HSC。我们将从三个具体方面完成总体目标 目标。在目标 1 中，我们将优化在肝脏中传递抗 miR-155 的 C-PEI-miRNA 纳米颗粒的配方 纤维化。基于我们的初步研究中鼓励抗纤维化活性，我们假设 Polyplex 用甘露糖和稳定胆固醇部分进行修饰将导致 miRNA 的有效递送 到活化的 KC，而过量的游离 C-PEI 将靶向活化的 HSC。在目标 2 中，我们将测试体内治疗 复合物在胆管结扎（BDL）模型和慢性酒精小鼠模型中的功效 暴露于 CCl4 的给药。目的是对治疗效果进行综合评价 在不同模型和纤维化的多个阶段中优化的 C-PEI-Chol/抗 miR-155 复合物。这 小鼠模型的研究结果将在培养的人类精密切割肝脏切片中得到验证。在目标 3 中，我们将 确定 miR-155 和 CXCR4 联合抑制的抗纤维化活性机制。虽然我们的 初步数据与 KC 中 miR-155 下调和活化 HSC 中 CXCR4 抑制一致， C-PEI/抗 miR-155 复合物的确切作用机制尚不清楚。因此，研究中 这一目标旨在确定复合物的潜在作用机制。总体来说，这个项目 将有助于从根本上理解 CXCR4 和 miR-155 的作用和治疗潜力 抑制 AALD 并将创新性地解决药物/miRNA 组合的靶向递送作为抗纤维化的问题 治疗。