Deciphering the internalization mechanism of WRAP:siRNA nanoparticles - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biochimica et Biophysica Acta:Biomembranes Année : 2020

Deciphering the internalization mechanism of WRAP:siRNA nanoparticles

Résumé

Gene silencing mediated by double-stranded small interfering RNA (siRNA) has been widely investigated as a potential therapeutic approach for a variety of diseases and, indeed, the first therapeutic siRNA was approved by the FDA in 2018. As an alternative to the traditional delivery systems for nucleic acids, peptide-based nanoparticles (PBNs) have been applied successfully for siRNA delivery. Recently, we have developed amphipathic cell-penetrating peptides (CPPs), called WRAP allowing a rapid and efficient siRNA delivery into several cell lines at low doses (20 to 50 nM). In this study, using a highly specific gene silencing system, we aimed to elucidate the cellular uptake mechanism of WRAP:siRNA nanoparticles by combining biophysical, biological, confocal and electron microscopy approaches. We demonstrated that WRAP:siRNA complexes remain fully active in the presence of chemical inhibitors of different endosomal pathways suggesting a direct cell membrane translocation mechanism. Leakage studies on lipid vesicles indicated membrane destabilization properties of the nanoparticles and this was supported by the measurement of WRAP:siRNA internalization in dynamin triple-KO cells. However, we also observed some evidences for an endocytosis-dependent cellular internalization. Indeed, nanoparticles co-localized with transferrin, siRNA silencing was inhibited by the scavenger receptor A inhibitor Poly I and nanoparticles encapsulated in vesicles were observed by electron microscopy in U87 cells. In conclusion, we demonstrate here that the efficiency of WRAP:siRNA nanoparticles is mainly based on the use of multiple internalization mechanisms including direct translocation as well as endocytosis-dependent pathways.

Domaines

Biotechnologies
Fichier principal
Vignette du fichier
Deshayes_Manuscript_SI.pdf (1.19 Mo) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-03007561 , version 1 (08-12-2020)

Identifiants

Citer

Sébastien Deshayes, Karidia Konate, Marion Dussot, Bérengère Chavey, Anaïs Vaissière, et al.. Deciphering the internalization mechanism of WRAP:siRNA nanoparticles. Biochimica et Biophysica Acta:Biomembranes, 2020, 1862 (6), pp.183252. ⟨10.1016/j.bbamem.2020.183252⟩. ⟨hal-03007561⟩
54 Consultations
179 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More