Targeted delivery of anti-microRNA-21 oligonucleotides using chlorotoxin-engineered extracellular vesicles for treatment of glioblastoma

Abstract

Glioblastoma is a devastating disease with a high mortality rate. Gene therapy with anti-microRNA inhibitors has been suggested as a new modality for treatment of glioblastoma. In this study, glioblastoma-targeted extracellular vesicles (EVs) were produced with specific ligands and evaluated as a carrier of anti-microRNA-21 oligonucleotides (AMO21). Angiopep-2 (ANG) and chlorotoxin (CTX) were linked to EVs by DNA recombination techniques. Cholesterol-conjugated AMO21 (AMO21c) was loaded onto the EVs decorated with ANG or CTX (ANG-EV or CTX-EV) by hydrophobic interactions. In vitro cellular uptake assays indicated that CTX-EV had higher delivery efficiency than unmodified EV (Unmod-EV) and ANG-EV. In addition, CTX-EV had higher transcytosis efficiency than the other EVs, suggesting that it effectively passes through the blood–brain barrier. In orthotopic glioblastoma animal models, CTX-EV delivered AMO21c more efficiently than Lipofectamine/AMO21c, Unmod-EV/AMO21c and ANG-EV/AMO21c. As a result, a greater decrease in tumour size with CTX-EV/AMO21c was observed, as compared with Lipofectamine/AMO21c, Unmod-EV/AMO21c and ANG-EV/AMO21c. CTX-EV/AMO21c induced the expression of the phosphatase and tensin homolog (PTEN) and programmed cell death 4 (PDCD4) genes in tumours. In addition, apoptosis levels in tumour tissues were enhanced by CTX-EV/AMO21c, as compared with the other samples. In conclusion, CTX-EV is effective for targeted delivery of AMO21 to glioblastoma and may have potential in glioblastoma gene therapy.