Aptamer-Driven Toxin Gene Delivery in U87 Model Glioblastoma Cells.

Abstract

A novel suicide gene therapy approach was tested in U87 MG glioblastoma multiforme cells. A 26nt G-rich double-stranded DNA aptamer (AS1411) was integrated into a vector at the 5' of a mammalian codon-optimized saporin gene, under CMV promoter. With this plasmid termed "APTSAP", the gene encoding ribosome-inactivating protein saporin is driven intracellularly by the glioma-specific aptamer that binds to cell surface-exposed nucleolin and efficiently kills target cells, more effectively as a polyethyleneimine (PEI)-polyplex. Cells that do not expose nucleolin at the cell surface such as 3T3 cells, used as a control, remain unaffected. Suicide gene-induced cell killing was not observed when the inactive saporin mutant SAPKQ DNA was used in the (PEI)-polyplex, indicating that saporin catalytic activity mediates the cytotoxic effect. Rather than apoptosis, cell death has features resembling autophagic or methuosis-like mechanisms. These main findings support the proof-of-concept of using PEI-polyplexed APTSAP for local delivery in rat glioblastoma models.

• Authors: Luana di Leandro, Francesco Giansanti, Sabrina Mei, Sara Ponziani, Martina Colasante, Matteo Ardini, Francesco Angelucci, Giuseppina Pitari, Michele d'Angelo, Annamaria Cimini, Maria Serena Fabbrini, Rodolfo Ippoliti
• Journal: Frontiers in pharmacology (Front Pharmacol) Vol. 12 Pg. 588306 ( 2021) ISSN: 1663-9812 [Print] Switzerland
• PMID: 33935695 (Publication Type: Journal Article)
• Copyright: Copyright © 2021 di Leandro, Giansanti, Mei, Ponziani, Colasante, Ardini, Angelucci, Pitari, d’Angelo, Cimini, Fabbrini and Ippoliti.