Novel anti-VEGF chimeric molecules delivered by AAV vectors for inhibition of retinal neovascularization.
Vascular endothelial growth factor (VEGF) is important in pathological neovascularization, which is a key component of diseases such as the wet form of age-related macular degeneration, proliferative diabetic retinopathy and cancer. One of the most potent naturally occurring VEGF binders is VEGF receptor Flt-1. We have generated two novel chimeric VEGF-binding molecules, sFLT01 and sFLT02, which consist of the second immunoglobulin (IgG)-like domain of Flt-1 fused either to a human IgG1 Fc or solely to the CH3 domain of IgG1 Fc through a polyglycine linker 9Gly. In vitro analysis showed that these novel molecules are high-affinity VEGF binders. We have demonstrated that adeno-associated virus serotype 2 (AAV2)-mediated intravitreal gene delivery of sFLT01 efficiently inhibits angiogenesis in the mouse oxygen-induced retinopathy model. There were no histological observations of toxicity upon persistent ocular expression of sFLT01 for up to 12 months following intravitreal AAV2-based delivery in the rodent eye. Our data suggest that AAV2-mediated intravitreal gene delivery of our novel molecules may be a safe and effective treatment for retinal neovascularization.
Pechan
P
P
Department of Molecular Biology, Genzyme Corporation, Framingham, MA 01701, USA.
Rubin
H
H
Lukason
M
M
Ardinger
J
J
DuFresne
E
E
Hauswirth
W W
WW
Wadsworth
S C
SC
Scaria
A
A
D000818:Animals; D000229:Dependovirus / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D012160:Retina / Q000378:metabolism*; D015861:Retinal Neovascularization / Q000378:metabolism / Q000628:therapy*; D014161:Transduction, Genetic / Q000379:methods; D019076:Transgenes; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics
D007141:Immunoglobulin Fc Fragments; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D042461:Vascular Endothelial Growth Factor A; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1