Delivery of anti-microRNA-21 antisense-oligodeoxynucleotide using amphiphilic peptides for glioblastoma gene therapy

Abstract

Inhibition of microRNA-21 (miR-21) has been shown to promote apoptosis of cancer cells and to reduce tumor size in glioblastoma. However, efficient carriers for antisense-oligodeoxynucleotide (antisense-ODN) against miR-21 have not yet been developed. In this study, the R3V6 peptide (R3V6) was evaluated as a carrier of antisense-ODN. In a gel retardation assay, R3V6 formed a complex with an antisense-ODN. The serum stability assay showed that R3V6 protected it from nucleases more efficiently than polyethylenimine (PEI; 25 kDa, PEI25k). A Renilla luciferase gene with a 3'-untranslated region (3'-UTR) recognizable by miR-21 (psiCHECK2-miR-21-UTR) was constructed for the antisense-ODN assay. psiCHECK2-miR-21-UTR expressed less Renilla luciferase in the cells with a higher level of miR-21 due to the effect of miR-21. In an in vitro transfection assay, the R3V6 peptide delivered anti-miR-21 antisense-ODN into cells more efficiently than PEI (25 kDa, PEI25k) and lipofectamine. As a result, antisense-ODN/R3V6 complex inhibited miR-21 and increased Renilla luciferase expression more efficiently than antisense-ODN/PEI25k or antisense-ODN/Lipofectamine complexes in both C6 and A172 glioblastoma cells. Furthermore, the antisense-ODN/R3V6 complexes reduced the level of miR-21 and induced apoptosis of glioblastoma cells. These results suggest that the R3V6 peptide may be a useful carrier of antisense-ODN for glioblastoma gene therapy.

Nanotechnology based approaches for detection and delivery of microRNA in healthcare and crop protection

“…Oh et al studied the effect of R3V6 peptide mediated delivery of anti-miR-21 in glioblastoma in animals [ 146 ]. Song et al studied the effect of peptide mediated delivery of anti-miR-21 in glioblastoma [ 147 ]. Suh et al studied the anti-cancer effects of peptide mediated delivery of miR-29b [ 148 ].…”

The role of micro-RNA in the regulation of signal pathways in gliomas

“…Так, введение с помощью стереотаксической хирургии олигонуклеотида anti-let-7 ксенографтным животным с глиобластомой показало, что наиболее эффективными способами введения являются интратуморальный и интравентрикулярный способы [133]. Одно из исследований [134] показало, что пептиды R3V6 способны защищать олигонуклеотиды anti-miR-21 от воздействия нуклеаз, при этом anti-miR-21 доставляются в клетку гораздо эффективнее, чем при других модификациях. Применение препарата снижало уровень miR-21 и индуцировало апоптоз в клетках глиобластомы, свидетельствуя о том, что R3V6 может служить "носителем" антисенс-олигонуклеотидов [134].…”

“…Одно из исследований [134] показало, что пептиды R3V6 способны защищать олигонуклеотиды anti-miR-21 от воздействия нуклеаз, при этом anti-miR-21 доставляются в клетку гораздо эффективнее, чем при других модификациях. Применение препарата снижало уровень miR-21 и индуцировало апоптоз в клетках глиобластомы, свидетельствуя о том, что R3V6 может служить "носителем" антисенс-олигонуклеотидов [134].…”

Nanoparticle-complexed antimiRs for inhibiting tumor growth and metastasis in prostate carcinoma and melanoma

“…For antimiR delivery, PEIs have been explored mainly as one component for modifying inorganic nanostructures like Au nanocages [ 26 , 27 ], in co-polymers or in multifunctional nanocomplexes [ 28 – 31 ]. While PEG-shielded carboxymethyl PEI nanogels [ 32 ] or disulphide-crosslinked PEIs [ 33 ] have been described as well, in another study the branched 25 kDa PEI “gold standard” performed only poorly in terms of complex stability, efficacy and biocompatibility [ 34 ].…”