Activated polyamidoamine dendrimers, a non-viral vector for gene transfer to the corneal endothelium.

Journal article

We investigated the efficiency of activated polyamidoamine dendrimers, a new class of nonviral vectors, to transfect rabbit and human corneas in ex vivo culture. In addition to assessing the expression of a marker gene we have demonstrated that this approach can be used to induce the production of TNF receptor fusion protein (TNFR-Ig), a protein with therapeutic potential. Whole thickness rabbit or human corneas were transfected ex vivo with complexes consisting of dendrimers and plasmids containing lacZ or TNFR-Ig genes. Following optimisation 6-10% of the corneal endothelial cells expressed the marker gene. Expression was restricted to the endothelium and was maximal after transfection with 18:1 (w/w) activated dendrimer:plasmid DNA ratio and culture for 3 days. The supernatant of corneas transfected with TNFR-Ig plasmid contained TNFR-Ig protein which was able to inhibit TNF-mediated cytotoxicity in a bioassay. We have therefore shown that activated dendrimers are an efficient nonviral vector capable of transducing corneal endothelial cells ex vivo. They may have applications in gene-based approaches aimed at prevention of corneal allograft rejection or in treatment of other disorders of corneal endothelium.

Authors: Hudde, T; Rayner, SA; Comer, R; Weber, M; Isaacs, J

• Publication status: Published
• Journal: Gene therapy
• Volume: 6
• Issue: 5
• Pages: 939-943
• Publication date: 1999-05-01
• DOI: 10.1038/sj.gt.3300886
• EISSN: 1476-5462
• ISSN: 0969-7128
• Language: English
• Keywords: Antigens, CD; Transfection; Receptors, Tumor Necrosis Factor; Endothelium, Corneal; Immunoglobulin Fc Fragments; Culture Techniques; Genetic Vectors; Time Factors; Immunoglobulin G; Recombinant Fusion Proteins; Receptors, Tumor Necrosis Factor, Type I; Rabbits; Humans; Animals