Aptamer-Driven Toxin Gene Delivery in U87 Model Glioblastoma Cells

• Main Authors: Luana di Leandro (Author), Francesco Giansanti (Author), Sabrina Mei (Author), Sara Ponziani (Author), Martina Colasante (Author), Matteo Ardini (Author), Francesco Angelucci (Author), Giuseppina Pitari (Author), Michele d'Angelo (Author), Annamaria Cimini (Author), Maria Serena Fabbrini (Author), Rodolfo Ippoliti (Author)
• Format: Book
• Published: Frontiers Media S.A., 2021-04-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

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.

Item Description

1663-9812
10.3389/fphar.2021.588306