Publication:
Intracellular trafficking and functional monitoring of miRNA delivery in glioblastoma using lipopolyplexes and the miRNA-ON RILES reporter system.

dc.contributor.authorSimion, Viorel
dc.contributor.authorHenriet, Elodie
dc.contributor.authorJuric, Viktorija
dc.contributor.authorAquino, Ruth
dc.contributor.authorLoussouarn, Claire
dc.contributor.authorLaurent, Yoan
dc.contributor.authorMartin, Francisco
dc.contributor.authorMidoux, Patrick
dc.contributor.authorGarcion, Emmanuel
dc.contributor.authorPichon, Chantal
dc.contributor.authorBaril, Patrick
dc.date.accessioned2023-02-09T09:39:19Z
dc.date.available2023-02-09T09:39:19Z
dc.date.issued2020-08-24
dc.description.abstractMicroRNA (miRNA) oligonucleotides therapeutics are potent and attractive drugs for cancer treatment, but the kinetics of their intracellular trafficking, RISC processing and interaction with their mRNA targets in the cells are still not well understood. Moreover, the absence of efficient carriers impairs their translation into the clinic. Here, we compare the kinetics of miRNA-133a activity after transfection of U87MG glioblastoma cells with either a home-made lipopolyplexes (LPRi) or with the RNAiMax transfection reagent. For this purpose, we combined miRNA intracellular trafficking studies by confocal microscopy with our previously described RILES miRNA-ON reporter system subcloned here in a lentivirus expression vector (LentiRILES) for longitudinal analysis of miRNA activity in transfected cells. Using the LentiRILES system, we report significant differences in terms of miRNA delivery kinetics performed by these two transfection regents. We decipher the mechanisms of miRNA delivery by LPRi and investigate the main steps of miRNA internalization and cytosolic processing. We demonstrate that LPRi preferentially uses caveolae-mediated endocytosis as the main internalization pathway, releases miRNA into the cytosol after the first 3 h of incubation, and addresses the cytosolic miRNAs to P-bodies, while a fraction of miRNAs are exported to the extracellular space through exosomes which were found fully capable to re-transfect the cells. We implanted the LentiRILES cells in the brain of mice and infused the tumours with LPRi.miRNA using the convection-enhanced delivery method. Bioluminescence imaging of the live mice revealed efficient delivery of miRNAs in glioblastoma tumours, attesting successful miRNA uptake, internalization and RISC activation in vivo. Overall, our study provides a comprehensive overview of miRNA intracellular trafficking and processing in a glioblastoma context and highlights the potential use of LPRi for miRNA-based therapy.
dc.identifier.doi10.1016/j.jconrel.2020.08.028
dc.identifier.essn1873-4995
dc.identifier.pmid32853728
dc.identifier.unpaywallURLhttps://www.sciencedirect.com/science/article/am/pii/S0168365920304685
dc.identifier.urihttp://hdl.handle.net/10668/16162
dc.journal.titleJournal of controlled release : official journal of the Controlled Release Society
dc.journal.titleabbreviationJ Control Release
dc.language.isoen
dc.organizationCentro Pfizer-Universidad de Granada-Junta de Andalucía de Genómica e Investigación Oncológica-GENYO
dc.page.number429-443
dc.pubmedtypeJournal Article
dc.pubmedtypeResearch Support, Non-U.S. Gov't
dc.pubmedtypeReview
dc.rights.accessRightsopen access
dc.subjectGlioblastoma
dc.subjectIntracellular trafficking
dc.subjectLipopolyplexes
dc.subjectP-bodies
dc.subjectmiRNA monitoring system
dc.subjectmicroRNA
dc.subject.meshAnimals
dc.subject.meshEndocytosis
dc.subject.meshExosomes
dc.subject.meshGlioblastoma
dc.subject.meshMice
dc.subject.meshMicroRNAs
dc.subject.meshTransfection
dc.titleIntracellular trafficking and functional monitoring of miRNA delivery in glioblastoma using lipopolyplexes and the miRNA-ON RILES reporter system.
dc.typeresearch article
dc.type.hasVersionAM
dc.volume.number327
dspace.entity.typePublication

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