Phosphorothioate oligonucleotides induction into experimental choroidal neovascularization by HVJ-liposome system.
OBJECTIVE
The purpose of this study was to determine whether the inactivated hemagglutinating virus of Japan (HVJ)-liposome method can induce phosphorothioate oligonucleotides effectively into an experimentally-induced choroidal neovascularization of rats. We also examined whether antisense phosphorothioate oligonucleotides against VEGF could be induced into choroidal neovascularization as a therapeutic agent by the HVJ-liposome method.


METHODS
The experiments were conducted on a rat model of choroidal neovascularization. FITC-labeled phosphorothioate oligonucleotides were coencapsulated in liposomes. The liposomes were coated with the envelope of inactivated HVJ and injected into the vitreous cavity following photocoagulation of pigmented rat eyes. The eyes were removed following injection, fixed, frozen and cut into thin sections. Induction of oligonucleotides was observed under a laser confocal scanning microscope for fluorescence and the development of choroidal neovascularization was evaluated histopathologically.


RESULTS
Phosphorothioate oligonucleotides were effectively induced into ganglion cells and into the cells of the choroidal neovascularization induced by laser photocoagulation. Highly effective induction of oligos was observed 3 to 14 days after intravitreal injection of HVJ-liposomes after which the level decreased. Antisense oligonucleotides against VEGF were induced specifically into cells in the choroidal neovascularization, however neovascularization was still observed.


CONCLUSIONS
Phosphorothioate oligonucleotides can be effectively induced into ganglion cells, and specifically into cells in choroidal neovascularization. Although antisense oligonucleotides against VEGF failed to prevent choroidal neovascularization, the HVJ-liposome method provided a highly effective means of inducing antisense oligos for in vivo antisense therapy.
Ogata
N
N
Department of Ophthalmology, Kansai Medical University, Moriguchi, Osaka, Japan.
Otsuji
T
T
Matsushima
M
M
Kimoto
T
T
Yamanaka
R
R
Takahashi
K
K
Wada
M
M
Uyama
M
M
Kaneda
Y
Y
D000818:Animals; D002829:Choroid / Q000098:blood supply*; D004337:Drug Carriers; D016228:Endothelial Growth Factors / Q000235:genetics; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008081:Liposomes; D008222:Lymphokines / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000473:pathology*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000494:pharmacology; D051381:Rats; D011914:Rats, Inbred BN; D018108:Respirovirus / Q000235:genetics; D013873:Thionucleotides / Q000235:genetics*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D004337:Drug Carriers; D016228:Endothelial Growth Factors; D005456:Fluorescent Dyes; D008081:Liposomes; D008222:Lymphokines; D016376:Oligonucleotides, Antisense; D013873:Thionucleotides; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D016650:Fluorescein-5-isothiocyanate