In vivo gene transfer into choroidal neovascularization by the HVJ liposome method.
OBJECTIVE
To evaluate the efficacy of the HVJ liposome method for gene transfer in rats with experimentally induced choroidal neovascularization.


METHODS
Plasmid DNA containing the LacZ reporter gene, or fluorescein isothiocyanate (FITC)-labeled double-stranded phosphorothioate oligodeoxynucleotides (S-ODNs), was encapsulated in liposomes. The liposomes were coated with the envelope of inactivated hemagglutinating virus of Japan (HVJ). Intense laser burns were applied to the posterior pole of the retina of pigmented rats to induce choroidal neovascularization. Following photocoagulation, HVJ liposome suspension was injected into the vitreous. On days 3, 7, 14, and 28 after injection, the eyes were removed and fixed. The eyes injected with LacZ gene were reacted with X-gal, frozen, and cut into thin sections. The sections were examined for the expression of the LacZ gene by light microscopy. The enucleated eyes injected with double-stranded S-ODNs were frozen, cut into thin sections, and examined a confocal scanning laser microscope for FITC labeling. Eyes without injection of HVJ liposomes served as controls.


RESULTS
Expression of LacZ genes (beta-galactosidase activity), or localization of FITC labeling, was observed mainly in the laser-induced choroidal neovascular tissue from 3 to 28 days after the intravitreal injection of HVJ liposome.


CONCLUSIONS
We conclude that the HVJ liposome method achieved effective gene transfer into choroidal neovascular tissue. Thus, this method can be used as a nonviral gene therapy system for the treatment of choroidal neovascularization in vivo.
Otsuji
T
T
Department of Ophthalmology, Kansai Medical University, Osaka, Japan. otsuji@takii.kmu.ac.jp
Ogata
N
N
Takahashi
K
K
Matsushima
M
M
Uyama
M
M
Kaneda
Y
Y
D000818:Animals; D020256:Choroidal Neovascularization / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000235:genetics; D004195:Disease Models, Animal; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D007763:Lac Operon / Q000235:genetics; D008081:Liposomes; D018613:Microscopy, Confocal; D051381:Rats; D011914:Rats, Inbred BN; D018108:Respirovirus / Q000235:genetics*; D013873:Thionucleotides / Q000235:genetics; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D004279:DNA, Viral; D008081:Liposomes; D013873:Thionucleotides; D001616:beta-Galactosidase