Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production

Bibliographic Details
Main Author:	Li, Zhefeng
Language:	English
Published:	The Ohio State University / OhioLINK 2020
Subjects:	Pharmacy Sciences
Online Access:	http://rave.ohiolink.edu/etdc/view?acc_num=osu1590062022556383

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spelling	ndltd-OhioLink-oai-etd.ohiolink.edu-osu15900620225563832021-08-03T07:15:01Z Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production Li, Zhefeng Pharmacy Sciences Extracellular Vesicles (EVs) represent a broad type of endogenous lipid vesicles carrying protein, peptide and nucleic acid derived from cells. Exosomes, a subtype of EVs, have shown promising potential as drug delivery vehicle, particularly for nucleic acid therapeutics. However, major challenges still remained for translational application including: 1) short of cell specific targeting limits the efficacy and raises off-target concern; 2) therapeutic value depends on heterogeneous cargo composition; 3) low productivity and high consumption in resources and time; etc. RNA nanotechnology is to study, design, and manipulate the nanometer-scale RNA architectures. The bottom-up self-assembly arrow-shape 3WJ RNA nanoparticles are homogenous and uniformed in size and shape, that can harbor different functionalities while retaining their tertiary folding and independent functionalities both in vitro and in vivo. This flexible platform using RNA nanotechnology to achieve tumor-specific targeting has been demonstrated over the last decade. Here we introduce a strategy to take advantage of both exosomes and RNA nanotechnology to develop a versatile platform for efficient target-specific delivery of RNA therapeutics for cancer treatment. We conjugated membrane-anchoring cholesterol at the tail of the arrow-shape 3WJ RNA nanoparticle to display RNA aptamer or folate on the surface of the exosomes, for enhancement of cancer cell binding and uptake. SiRNA targeting BIRC5 gene, which is also known as survivin gene, was used as therapeutic cargo for proof of concept. The high cost and low yield of exosomes derived from traditional cell culture was one of the blockages for therapeutic application. We took two different strategies to address the exosome productivity question. 1) We established the ginger derived exosome-like nanovesicles (GDENs) production platform using eatable plant as alternative sources for exosomes. Purified GDENs had similar size, density, and morphology to HEK293T cell derived exosomes. We observed inhibition of tumor growth on a xenograft model by intravenous administration of survivin siRNA encapsulated by folate displaying GDENs, which reveals the potential of GDENs as an economic delivery system for siRNA. 2) We used Hollow-fiber bioreactor coupled with tangential flow filtration (TFF) system to build a workflow for HEK293T cell exosome production that improved the overall efficiency for 100 times.Endosomal entrapment is one of the most challenging barriers for delivering RNAi therapeutics. We chose folate as example to study how the exosomes platform can facilitate cytosol delivery. Folate receptor is a cell surface glycoprotein that overexpresses in many cancer cells. Folate is a high affinity ligand for targeting cancer cell but has low efficiency for delivering RNAi therapeutic due to endosome trapping. We demonstrated that the efficient cancer suppression with the folate-displaying exosome was due to the receptor-mediated cytosol delivery of the siRNA payload through direct fusion, as attested by fluorescence colocalization analysis, gene knockdown assay and tumor regression. In conclusion, we had developed a siRNA delivery platform of ligand displaying exosomes by RNA nanoparticles for cancer treatment with efficient cytosolic delivery. 2020-11-13 English text The Ohio State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=osu1590062022556383 http://rave.ohiolink.edu/etdc/view?acc_num=osu1590062022556383 restricted--full text unavailable until 2022-08-25 This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection	NDLTD
language	English
sources	NDLTD
topic	Pharmacy Sciences
spellingShingle	Pharmacy Sciences Li, Zhefeng Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production
author	Li, Zhefeng
author_facet	Li, Zhefeng
author_sort	Li, Zhefeng
title	Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production
title_short	Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production
title_full	Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production
title_fullStr	Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production
title_full_unstemmed	Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production
title_sort	ligand displaying exosomes by arrowtail rna nanoparticles for sirna delivery and scale-up production
publisher	The Ohio State University / OhioLINK
publishDate	2020
url	http://rave.ohiolink.edu/etdc/view?acc_num=osu1590062022556383
work_keys_str_mv	AT lizhefeng liganddisplayingexosomesbyarrowtailrnananoparticlesforsirnadeliveryandscaleupproduction
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Ligand Displaying Exosomes by Arrowtail RNA Nanoparticles for siRNA Delivery and Scale-up Production

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