[en] RNA interference via small interfering RNA (siRNA) offers promising potential for treating glioblastoma (GB), the most common and aggressive primary brain tumor. While local siRNA delivery could bypass the blood-brain barrier and achieve high drug concentrations, challenges remain in developing effective delivery systems that ensure siRNA stability, efficient cellular uptake, endolysosomal escape, and long-term brain availability. The Argonaute-2 (Ago2) protein, a core component of the RNA interference machinery, naturally binds siRNA and protects it from degradation. Ago2 is also found in biological fluids, such as blood, and in extracellular vesicles, including exosomes and microvesicles, making it a potential natural vector for siRNA delivery. This study explores Ago2 as a natural vector for siRNA in nanoparticle-based delivery systems to enhance siRNA transport and gene silencing in GB. Neutral polymeric siRNA/Ago2-loaded nanoparticles (NPs), around 300 nm in size, were formulated with high encapsulation efficiencies (85.1 % for protein and 57.5 % for siRNA). In vitro testing in U87MG GB cells stably expressing luciferase (U87MG Bml1) showed that, in contrast to free siRNA/Ago2 complexes, siRNA/Ago2-loaded NPs effectively reduced luciferase expression compared to scramble siRNA formulations, with no significant cytotoxicity at 48 h. Interestingly, the introduction of exogenous Ago2 into cells unexpectedly upregulated luciferase expression, suggesting that Ago2 may also influence global gene expression beyond its role in siRNA-mediated silencing. These findings highlight the potential of siRNA/Ago2-loaded NPs for local siRNA delivery in GB and suggest that Ago2 could play a broader regulatory role in gene expression, in addition to enhancing siRNA activity. Further investigation is needed to explore Ago2's impact on gene regulation and its potential as a therapeutic tool in GB treatment, particularly in combination with biocompatible hydrogels for sustained delivery.
Disciplines :
Chemistry
Author, co-author :
Rinaldi, A.; Université d’ Angers, Inserm U1307, CNRS U6075, Nantes Université, CRCI2NA, Angers, France ; Università degli Studi di Modena e Reggio Emilia, Modena, Italy
Nazir, M. Louis Fathy; Université d’ Angers, Inserm U1307, CNRS U6075, Nantes Université, CRCI2NA, Angers, France
Barbotin, M.; Université d’ Angers, Inserm U1307, CNRS U6075, Nantes Université, CRCI2NA, Angers, France
Roy, C.; Université d’ Angers, Inserm U1307, CNRS U6075, Nantes Université, CRCI2NA, Angers, France ; PRIMEX (Plateforme de Radiobiologie et d’Imageries Expérimentales), SFR, Université d'Angers, Angers, France
Basset, L.; Université d’ Angers, Inserm U1307, CNRS U6075, Nantes Université, CRCI2NA, Angers, France ; Department of Pathology, University Hospital of Angers, Angers, France
Rousseau, A.; Université d’ Angers, Inserm U1307, CNRS U6075, Nantes Université, CRCI2NA, Angers, France ; Department of Pathology, University Hospital of Angers, Angers, France
Garcion, E. ; Université d’ Angers, Inserm U1307, CNRS U6075, Nantes Université, CRCI2NA, Angers, France ; PRIMEX (Plateforme de Radiobiologie et d’Imageries Expérimentales), SFR, Université d'Angers, Angers, France ; PACEM (Plateforme d’Analyse Cellulaire et Moléculaire), SFR, Université d'Angers, Angers, France
Università Italo Francese INCa - Institut National du Cancer LNCC - Ligue Nationale Contre le Cancer UA - Université d'Angers INSERM - Institut National de la Santé et de la Recherche Médicale ANR - Agence Nationale de la Recherche Cancéropôle du Grand Ouest Conseil Régional des Pays de la Loire
Funding text :
The work was supported by the Institut National de la Sant\u00E9 et de la Recherche M\u00E9dicale (INSERM), the University of Angers , the Institut National du Cancer ( INCa ) under the frame of the PL- BIO project MARENGO, the \u201CLigue Nationale contre le Cancer\u201D and the \u201CComit\u00E9 D\u00E9partemental de Maine-et-Loire de la Ligue contre le Cancer\u201D (CD49) under the frame of the FUSTARG project. It was additionally related to: the French National Agency for Research (ANR) under the frame of EuroNanoMed III (project GLIOSILK) [ANR-19-ENM3-0003-01]) and of the \u201CFrance 2030 Investment Plan\u201D Labex Iron [ANR-11-LABX-18- 01], the \u201CR\u00E9gion Pays-de-la-Loire\u201D under the frame of the TARGET\u2019IN project and the \u201CTumour targeting, imaging and radio-therapies network\u201D of the \u201CCanc\u00E9rop\u00F4le Grand-Ouest\u201D (France). A. Rinaldi was a PhD fellow from the Program VINCI 2020 - Universit\u00E9 Franco Italienne (project number: C3-1419, \u201CNovel nanotechnological approaches for glioblastoma targeting\u201D). Finally, the authors thank R. Mallet, R. Perrot and F. Manero from the SCIAM (Service Commun d\u2019Imagerie et d\u2019Analyses Microscopiques, Angers, France) for SEM and TEM analyses. We gratefully acknowledge that this publication contributed to a PhD thesis awarded the UFI 2025 Cotutelle PhD Prize by the Universit\u00E9 Franco-Italienne. We thank the UFI for their support and kindly refer readers to their website https://www.universite-franco-italienne.org/fr/ for more information.
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