Fruit exosome-like particles for therapeutic delivery of extracellular miRNAs

用于治疗性递送细胞外 miRNA 的水果外泌体样颗粒

基本信息

批准号：
8962196
负责人：
PEIXUAN GUO
金额：
$ 66.71万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8962196
关键词：
Adverse effects Animals Antibodies Biohazardous Substance Biological Assay Brain Brain Neoplasms Breast Cells Clinic Clinical Colon Carcinoma Colonic Neoplasms Curcumin DNA Data Dendritic Cells Development Disease Dose Down-Regulation Edible Plants Encapsulated Environment Evaluation Folic Acid Fruit Gene Expression Goals Grapefruit Grapes Health Human Immune response Immunotherapy In Vitro Inflammatory Intranasal Administration Knock-in Mouse Knock-out Laboratories Light Lipids Liposomes Literature Liver MHC Class I Genes Malignant Neoplasms Mammalian Cell Metastatic Neoplasm to the Liver MicroRNAs Modeling Mus Myelogenous Myeloid Cells NK Cell Activation Neoplasm Metastasis Non-Viral Vector Nucleic Acids Oral Administration Paclitaxel Pharmaceutical Preparations Phenotype Play Prevention Production Proteins Protocols documentation Publishing RNA Research Personnel Role Safety Science Signal Transduction Small Interfering RNA Small RNA Specificity Suppressor-Effector T-Lymphocytes System Testing Therapeutic Therapeutic Agents Tissues Tomatoes Toxic effect Tumor Tissue Viral Vector aqueous base cancer cell cancer therapy cell type chemotherapeutic agent chemotherapy cost expression vector extracellular in vivo intravenous administration killings large scale production migration mouse model nanoparticle nanosized nanovector neoplastic cell overexpression particle prevent receptor targeted delivery therapeutic miRNA tool trafficking tumor vector-induced

项目摘要

DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop exosome-like nanoparticles as a delivery vehicle for targetable delivery of extracellular RNA (exRNA) primarily focusing on miRNAs which modulate gene expression and induce immune responses upon delivery in mouse tumor models. Development of exRNA-based therapeutics must overcome challenges including issues of delivery, potential off-target effects, safety, toxicity, the cost of large scale production, elimination of potential biohazards to the environmen and tissue specificity. Unlike the situation with artificially synthesized nanoparticles or liposomes, naturally derived nanoparticle-size exosomes are released from many different types of cells. miRNAs and RNAs can be encapsulated in naturally derived exosomes and the RNAs are stable, and a number of miRNAs can be naturally encapsulated simultaneously in these exosomes. Recently, data published by ours and other groups have demonstrated that mammalian exosomes can deliver poorly soluble agents, chemotherapeutic drugs, and siRNA in vivo. A limitation of this mode is the need for large-scale production of exosomes, and potential biosafety. Our preliminary data show that exosome-like nanoparticles can be isolated in large quantities from the tissue of edible plants, including grapes. We have demonstrated that exosome-like nanoparticles from grapes are composed of small RNAs, proteins, and lipids. An edible plant-derived nano-vector (EPNV) assembled from grape exosome-like derived lipids is capable of encapsulating siRNAs. Co-delivery of folic acid, with the chemotherapy drug paclitaxel by EPNVs results in an increased efficiency in tumor tissue targeting. Reducing the size of EPNVs by passing them through an appropriate pore size filter significantly enhances EPNV translocation to the brain and oral administration of the EPNVs results in their migration to the liver. Based on these preliminary data and the current literature, we propose to: (1) Determine whether an EPNV can deliver therapeutic miR17 via oral administration to target metastatic colon tumor in the liver, subsequently inducing NK cell activation to kill metastatic tumor cells; (2) Determine whether miR155 encapsulated in EPNV promotes differentiation of myeloid derived suppressor cells (MDSCs) into mature dendritic cells in mouse breast/brain tumor models; (3) Determine whether a therapeutic dose of miRNA17 or miRNA155 delivered by EPNV induces no side-effects in mouse tumor models; and (4) Determine whether EPNV can be produced in large scale amounts economically for use in clinical settings. Demonstration of using a EPNV-based delivery system loaded with a desired therapeutic extracellular RNA (exRNA) including microRNA to target to inflammatory cells and tumor cells in vivo would not only be a significant step forward in the treatment of cancer, but also provide a powerful tool for identifying the role and effects of each small RNA encapsulated in resident or circulating microvesicles on the recipient cells. We will accomplish this in our study by knock-out and knock-in strategies.

描述（由申请人提供）：该提案的总体目标是开发外泌体样纳米粒子作为递送载体，用于靶向递送细胞外RNA（exRNA），主要关注在小鼠肿瘤中递送后调节基因表达并诱导免疫反应的miRNA模型。基于 exRNA 的疗法的开发必须克服挑战，包括递送问题、潜在的脱靶效应、安全性、毒性、大规模生产的成本、消除对环境的潜在生物危害和组织特异性。与人工合成的纳米颗粒或脂质体的情况不同，天然衍生的纳米颗粒大小的外泌体是从许多不同类型的细胞中释放的。 miRNA和RNA可以封装在天然来源的外泌体中，并且RNA是稳定的，并且许多miRNA可以同时自然封装在这些外泌体中。最近，我们和其他小组发表的数据表明，哺乳动物外泌体可以在体内递送难溶性药物、化疗药物和 siRNA。这种模式的局限性是需要大规模生产外泌体，并且存在潜在的生物安全性。我们的初步数据表明，可以从包括葡萄在内的可食用植物的组织中大量分离出外泌体样纳米颗粒。我们已经证明，来自葡萄的外泌体样纳米颗粒由小 RNA、蛋白质和脂质组成。由葡萄外泌体样衍生脂质组装而成的可食用植物源纳米载体 (EPNV) 能够封装 siRNA。 EPNV 联合递送叶酸和化疗药物紫杉醇可提高肿瘤组织靶向的效率。通过使 EPNV 通过适当孔径的过滤器来减小 EPNV 的大小，可显着增强 EPNV 向大脑的易位，并且口服给予 EPNV 会导致它们迁移到肝脏。基于这些初步数据和现有文献，我们建议：（1）确定EPNV是否可以通过口服给药将治疗性miR17递送至肝脏中的转移性结肠肿瘤，随后诱导NK细胞活化以杀死转移性肿瘤细胞； (2) 确定EPNV中封装的miR155是否促进小鼠乳腺/脑肿瘤模型中骨髓源性抑制细胞（MDSC）分化为成熟树突状细胞； (3) 确定EPNV递送的治疗剂量的miRNA17或miRNA155在小鼠肿瘤模型中是否不会引起副作用； (4) 确定是否可以经济地大规模生产 EPNV 以用于临床环境。演示使用基于 EPNV 的递送系统装载所需的治疗性细胞外 RNA (exRNA)，包括 microRNA，在体内靶向炎症细胞和肿瘤细胞，这不仅是癌症治疗方面的重要一步，而且还提供了强大的工具用于确定封装在常驻或循环微泡中的每种小 RNA 对受体细胞的作用和影响。我们将在我们的研究中通过敲除和敲入策略来实现这一目标。