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

• 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
• 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

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(申请人提供)：这项建议的总体目标是开发外周体样纳米颗粒作为靶向递送细胞外RNA(ExRNA)的载体，主要集中在miRNAs上，miRNAs在递送小鼠肿瘤模型时调节基因表达并诱导免疫反应。基于exRNA的疗法的发展必须克服包括传递、潜在的靶外效应、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等问题。与人工合成的纳米颗粒或脂质体不同，自然衍生的纳米颗粒大小的外切体可以从许多不同类型的细胞中释放出来。MiRNAs和RNAs可以被包裹在天然来源的外切体中，并且RNAs是稳定的，并且许多miRNAs可以自然地同时被包裹在这些外切体中。最近，我们和其他研究小组发表的数据表明，哺乳动物外切体可以在体内运送难溶的药物、化疗药物和siRNA。这种模式的一个局限性是需要大规模生产外切体，以及潜在的生物安全。我们的初步数据显示，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显子状纳米颗粒。我们已经证明了葡萄中的外切体样纳米颗粒是由小RNA、蛋白质和脂类组成的。由葡萄外切体样脂组装而成的可食用植物源纳米载体(EPNV)能够包裹siRNAs。EPNvs将叶酸与化疗药物紫杉醇联合应用，可提高肿瘤组织靶向的效率。通过适当的孔径过滤器减小EPNV的大小可以显著增强EPNV向大脑的移位，口服EPNV会导致它们向肝脏迁移。基于这些初步数据和当前的文献，我们建议：(1)确定EPNV能否通过口服给药来靶向肝脏转移的结肠癌，随后诱导NK细胞激活以杀死转移的肿瘤细胞；(2)确定EPNV包裹的miR155是否能促进小鼠乳腺/脑肿瘤模型中髓系来源的抑制细胞(MDSCs)向成熟树突状细胞的分化；(3)确定EPNV所传递的治疗性剂量的miRNA17或miRNA155是否在小鼠肿瘤模型中没有引起副作用；以及(4)确定是否可以经济地大规模生产EPNV用于临床。证明使用EPNV为基础的递送系统装载所需的治疗性细胞外RNA(ExRNA)(包括microRNA)来靶向体内的炎症细胞和肿瘤细胞，不仅是在癌症治疗方面向前迈出的重要一步，而且将为 确定驻留或循环中的微泡中包裹的每个小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出和敲入策略来实现这一点。