AUTHOR=Oshchepkova Anastasiya , Chernikov Ivan , Miroshnichenko Svetlana , Patutina Olga , Markov Oleg , Savin Innokenty , Staroseletz Yaroslav , Meschaninova Mariya , Puchkov Pavel , Zhukov Sergey , Kupryushkin Maxim , Maslov Mikhail , Sen’kova Aleksandra , Vlassov Valentin , Chernolovskaya Elena , Zenkova Marina TITLE=Extracellular vesicle mimetics as delivery vehicles for oligonucleotide-based therapeutics and plasmid DNA JOURNAL=Frontiers in Bioengineering and Biotechnology VOLUME=12 YEAR=2024 URL=https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2024.1437817 DOI=10.3389/fbioe.2024.1437817 ISSN=2296-4185 ABSTRACT=Introduction

Small membrane particles called extracellular vesicles (EVs) transport biologically active cargo between cells, providing intercellular communication. The clinical application of EVs is limited due to the lack of scalable and cost-effective approaches for their production and purification, as well as effective loading strategies.

Methods

Here we used EV mimetics produced by cell treatment with the actin-destabilizing agent cytochalasin B as an alternative to EVs for the delivery of therapeutic nucleic acids.

Results

Cytochalasin-B-inducible nanovesicles (CINVs) delivered a fully modified N-(methanesulfonyl)- or mesyl (µ-) antisense oligonucleotide to B16 melanoma cells, selectively decreasing the level of target microRNA-21 with effectiveness comparable to that observed upon Lipofectamine 2000-mediated delivery. The efficiency of the CINV-mediated delivery of plasmid DNA encoding EGFP varied depending on the type of recipient cells. Surprisingly, under experimental conditions, CINVs were unable to deliver both modified and natural short RNA duplexes—small interfering RNA and immunostimulatory RNA—probably due to their poor loading into CINVs.

Discussion

CINVs demonstrated unique properties for the delivery of therapeutic nucleic acids, especially for antisense oligonucleotide-based therapy.