AAV plasmid DNA simplifies liver-directed in vivo gene therapy: comparison of expression levels after plasmid DNA-, adeno-associated virus- and adenovirus-mediated liver transfection

✍ Scribed by Axel Doenecke; Alexander Krömer; Marcus N. Scherer; Hans-Jürgen Schlitt; Edward K. Geissler

• Publisher: John Wiley and Sons
• Year: 2010
• Tongue: English
• Weight: 208 KB
• Volume: 12
• Category: Article
• ISSN: 1099-498X
• DOI: 10.1002/jgm.1498

✦ Synopsis

Abstract

Background

Successful liver gene therapy depends on efficient gene transfer techniques and long‐lasting gene expression after successful transfer. Over the last decades, important progress has been made with the introduction of viral vectors using animal models, although their use is hampered by a complex and costly preparation compared to the simple and cost‐effective preparation of plasmid DNA. These problems become even more critical when considering the application of viral vectors in human gene therapy and gene therapy trials. In a previous study, we were able to show that the hydrodynamics‐based gene transfer of plasmid‐DNA, containing the adeno‐associated‐virus specific inverted terminal repeats (AAV‐ITR), prolongs gene expression in the liver, although it remained unclear whether plasmid gene transfer could achieve similar expression levels compared to viral‐vector gene transfer.

Methods

Rat livers were transfected in‐vivo with AAV‐ITR‐containing plasmid‐DNA using a modified hydrodynamics‐based procedure. Expression levels were monitored thereafter and compared with expression levels after viral‐vector gene transfer.

Results

A high and stable long‐term expression was achieved after in vivo transfection of rat livers with AAV‐ITR‐containing plasmids. The expression course resembled that after AAV‐mediated gene transfer, and the expression was at least as high, and lasted as long, compared to recombinant AAV‐mediated gene transfer.

Conclusions

We consider AAV‐ITR‐containing plasmids as a simple and cost‐effective alternative to recombinant viral vectors, especially for liver‐directed gene therapy in rodents. With ongoing progress in gene transfer methods for naked DNA, these plasmids may also become a successful alternative to recombinant viral vectors in human gene therapy. Copyright © 2010 John Wiley & Sons, Ltd.