Rapid ocular angiogenic control via naked DNA delivery to cornea.
OBJECTIVE
To determine the efficacy and safety of naked plasmid gene therapy to the corneal stroma and epithelium.


METHODS
Naked plasmid DNA was injected under pressure into the cornea of mice. The expression of genes coding for beta galactosidase (beta-gal), enhanced green fluorescent protein (EGFP), vascular endothelial growth factor (VEGF), and soluble Flt-1 (s-Flt) was recorded and measured with regard to dose, time course, and bioactivity.


RESULTS
LacZ gene expression of the protein beta-gal was demonstrated as early as 1 hour, with expression persisting for 10 days. Plasmid-injected corneas remained clear and free of inflammation. EGFP was bicistronically expressed with VEGF to demonstrate the practicality of simultaneous in vivo analysis of gene expression and growth factor bioactivity. Corneal injection of a plasmid containing VEGF cDNA induced corneal and anterior chamber neovascularization. Moreover, corneal injection of plasmid containing the cDNA for the soluble form of the VEGF receptor Flt-1 effectively prevented corneal neovascularization.


CONCLUSIONS
The cornea is readily accessible for gene therapy in the laboratory and in the clinic. The method described is safe, effective, titratable, and easily monitored. Naked DNA delivery to the cornea has the potential to alter the treatment of a wide variety of corneal and anterior segment diseases.
Stechschulte
S U
SU
The Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Joussen
A M
AM
von Recum
H A
HA
Poulaki
V
V
Moromizato
Y
Y
Yuan
J
J
D'Amato
R J
RJ
Kuo
C
C
Adamis
A P
AP
D000818:Animals; D003315:Cornea / Q000378:metabolism* / Q000473:pathology; D016510:Corneal Neovascularization / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D004247:DNA / Q000008:administration & dosage*; D004305:Dose-Response Relationship, Drug; D016228:Endothelial Growth Factors / Q000096:biosynthesis / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008222:Lymphokines / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection / Q000379:methods*; D042461:Vascular Endothelial Growth Factor A; D040281:Vascular Endothelial Growth Factor Receptor-1; D042442:Vascular Endothelial Growth Factors; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D016228:Endothelial Growth Factors; D008164:Luminescent Proteins; D008222:Lymphokines; D011518:Proto-Oncogene Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D049452:Green Fluorescent Proteins; D004247:DNA; D020794:Receptor Protein-Tyrosine Kinases; D040281:Vascular Endothelial Growth Factor Receptor-1; D001616:beta-Galactosidase