T. Hudde et al., Activated polyamidoamine dendrimers, a non-viral vector for gene transfer to the corneal endothelium, GENE THER, 6(5), 1999, pp. 939-943
We investigated the efficiency of activated polyamidoamine dendrimers, a ne
w class of nonviral vectors, to transfect rabbit and human corneas in ex vi
vo culture. In addition ib assessing the expression of a marker gene we hav
e demonstrated that this approach can be used to induce the pro; production
of TNF receptor fusion protein (TNFR-lg), a protein with therapeutic poten
tial. Whole thickness rabbit or human corneas were transected ex vivo with
complexes consisting of dendrimers and plasmids containing lacZ or TNFR-lg
genes. Following optimisation 6-10% of the corneal endothelial cells expres
sed the marker gene. Expression was restricted to the endothelium and was m
aximal after transfection with 18:1 (W/W) activated dendrimer:plasmid DNA r
atio and culture for 3 days, The supernatant of corneas transfected with TN
FR-lg plasmid contained TNFR-lg protein which was able to inhibit TNF-media
ted cytotoxicity in a bioassay We have therefore shown that activated dendr
imers are an efficient nonviral vector capble of transducing corneal endoth
elial cells ex vivo. They may have applications in gene-based approaches ai
med at prevention of corneal allograft rejection or in treatment of other d
isorders of corneal endothelium.