Inhibition of experimental choroidal neovascularization by overexpression of tissue inhibitor of metalloproteinases-3 in retinal pigment epithelium cells.
OBJECTIVE
To evaluate the feasibility of introducing exogenous tissue inhibitor of metalloproteinases-3 gene into the rat retinal pigment epithelium using hemagglutinating virus of Japan liposomes and to assess the effect of tissue inhibitor of metalloproteinases-3 overexpression in retinal pigment epithelium cells on the formation of experimental choroidal neovascularization.


METHODS
Hemagglutinating virus of Japan liposomes containing hemagglutin epitope-tagged tissue inhibitor of metalloproteinases-3 gene were injected into the subretinal space in rat eyes. Localization of oligonucleotides was evaluated by fluorescence microscopy. Exogenous tissue inhibitor of metalloproteinases-3 mRNA expression was assessed by reverse transcribed polymerase chain reaction. Exogenous tissue inhibitor of metalloproteinases-3 protein expression was visualized by immunostaining with monoclonal antibody 12CA5 against the hemagglutin epitope. Three days after transfection of tissue inhibitor of metalloproteinases-3 gene into retinal pigment epithelium cells, intense laser photocoagulation was performed and the incidence of choroidal neovascularization was assessed by fluorescein fundus angiography.


RESULTS
Exogenous tissue inhibitor of metalloproteinases-3 mRNA expression in the choroid and retina was detected on day 3. The efficiency of tissue inhibitor of metalloproteinases-3 gene transfection into retinal pigment epithelium cells was greatest on day 7 and decreased gradually thereafter. The incidence of choroidal neovascularization in tissue inhibitor of metalloproteinases-3 gene-transfected eyes was markedly decreased compared with controls.


CONCLUSIONS
This study shows that tissue inhibitor of metalloproteinases-3 gene can be transferred into rat retinal pigment epithelium using the hemagglutinating virus of Japan-liposome method and that tissue inhibitor of metalloproteinases-3 gene overexpression can inhibit development of experimental choroidal neovascularization. This method may represent a future treatment modality for human macular degeneration associated with choroidal neovascularization.
Takahashi
T
T
Department of Ophthalmology, Osaka University Medical School, Osaka, Japan. tadashi@vision.eri.harvard.edu
Nakamura
T
T
Hayashi
A
A
Kamei
M
M
Nakabayashi
M
M
Okada
A A
AA
Tomita
N
N
Kaneda
Y
Y
Tano
Y
Y
D000818:Animals; D002829:Choroid / Q000201:enzymology; D020256:Choroidal Neovascularization / Q000201:enzymology / Q000517:prevention & control*; D017931:DNA Primers / Q000737:chemistry; D005451:Fluorescein Angiography; D005654:Fundus Oculi; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D017075:Laser Coagulation; D008297:Male; D008856:Microscopy, Fluorescence; D010857:Pigment Epithelium of Eye / Q000201:enzymology*; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D011914:Rats, Inbred BN; D018108:Respirovirus / Q000235:genetics; D012160:Retina / Q000201:enzymology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D017931:DNA Primers; D012333:RNA, Messenger; D019717:Tissue Inhibitor of Metalloproteinase-3