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Subretinal Injection (subretinal + injection)
Selected AbstractsPigment epithelium-derived factor inhibits retinal and choroidal neovascularizationJOURNAL OF CELLULAR PHYSIOLOGY, Issue 2 2001Keisuke Mori In this study, we investigated whether overexpression of pigment epithelium-derived factor (PEDF) by gene transfer can inhibit neovascularization by testing its effect in three different models of ocular neovascularization. Intravitreous injection of an adenoviral vector encoding PEDF resulted in expression of PEDF mRNA in the eye measured by RT-PCR and increased immunohistochemical staining for PEDF protein throughout the retina. In mice with laser-induced rupture of Bruch's membrane, choroidal neovascularization was significantly reduced after intravitreous injection of PEDF vector compared to injection of null vector or no injection. Subretinal injection of the PEDF vector resulted in prominent staining for PEDF in retinal pigmented epithelial cells and strong inhibition of choroidal neovascularization. In two models of retinal neovascularization (transgenic mice with increased expression of vascular endothelial growth factor (VEGF) in photoreceptors and mice with oxygen-induced ischemic retinopathy), intravitreous injection of null vector resulted in decreased neovascularization compared to no injection, but intravitreous injection of PEDF vector resulted in further inhibition of neovascularization that was statistically significant. These data suggest that sustained increased intraocular expression of PEDF by gene therapy might provide a promising approach for treatment of ocular neovascularization. © 2001 Wiley-Liss, Inc. [source] Gene therapy for posterior uveitisACTA OPHTHALMOLOGICA, Issue 2009AD DICK Purpose To investigate the role of gene therapy incorporating release of immunomodulatory cytokines in animal models of intraocular inflammation Methods By inoculating with either AAV or lente viruses incorporating genes for IL-1RA or IL-10 into either the anterior chamber or subretinally we onserved the ability to suppress either endotoxin induced uveitis (EIU) or experimental autoimmiune uveoretinitis (EAU). Results Anterior chamber inoculation with lente-IL-10 or IL-1RA successfully suppresses inflammation and protein exudation into the eye during the course of EIU. Subretinal injection of AAV-IL-10 suppresses EAU. The extent of local macrophage activation is also suppressed as there is marked reduction in nitrotyrosine expression within the retina. Conclusion Gene therapy with immunomodulatory cytokines offers a potential to suppress active inflammatory processes within the retina. Mechanisms will be discussed in the talk in relation to macrophage activation and restoring myeloid cell (microgolial) homeostasis within the retina. [source] Comparison of wild-type and class I integrase mutant-FIV vectors in retina demonstrates sustained expression of integrated transgenes in retinal pigment epitheliumTHE JOURNAL OF GENE MEDICINE, Issue 12 2003Nils Loewen Abstract Background In neonatal and adult rodent retina, substantial lentiviral vector expression has been detected primarily in retinal pigment epithelium (RPE), except in very young animals (2,5 days post-natal). In non-retinal tissues, studies of lentiviral vectors have utilized various controls. Among the most stringent are class I integrase mutants, which selectively block the integration reaction while leaving all other gag/pol -encoded functions intact. For HIV-1 vectors injected into brain, these have been used to simultaneously control for pseudotransduction and verify that long-term expression requires integration. Such experiments compare particles that differ only in a single amino acid within a single enzyme that forms a very small molar fraction of the virion. Class I integrase mutants have not been described for feline immunodeficiency virus (FIV) integrase, or tested in the eye for any lentiviral vector. Methods We compared subretinally and intravitreally injected FIV vectors and followed animals for up to 7 months, a duration that exceeds prior studies. We also compared the wild-type (WT) vector with one incorporating a single class I amino acid mutation in FIV integrase (D66V). A mock vector (packaging construct absent) was an alternative control. All vectors were vesicular stomatitis virus glycoprotein G (VSV-G)-pseudotyped and were injected on day 7 of life. One group of animals received either subretinal or intravitreal injections of WT vector in the right eyes. Control left eyes were injected with mock vector. These animals were sacrificed at 2 or 7 days post-injection. A second group received subretinal injections of either WT vector or equivalent D66V vector (reverse transcriptase-normalized to WT), and were analyzed after 2, 3 and 7 months. All eyes were scored for marker gene (,-galactosidase) expression by an observer blinded to vector assignments. Results Subretinal FIV vector injections were much more effective than intravitreal injections. The RPE was the principal retinal layer transduced by the WT vector, and at least 50% of the area of the retina expressed the marker gene at 3 and 7 months. Occasional cells in inner retinal layers also expressed ,-galactosidase at these time points. The sustained retinal expression produced by subretinally injected vector was blocked by the D66V mutation. Conclusions These results show that class I integrase mutant FIV vectors are useful control vectors, and that VSV-G-pseudotyped FIV vectors produce extensive retinal expression for at least 215 days, the longest duration yet reported for lentiviral vectors in retina. Transgene expression is mostly restricted to RPE after post-natal day 7 in rats, suggesting that FIV vectors could be used to target RPE for gene therapy. Copyright © 2003 John Wiley & Sons, Ltd. [source] Feline immunodeficiency virus vectors.THE JOURNAL OF GENE MEDICINE, Issue 5 2002Gene transfer to mouse retina following intravitreal injection Abstract Background Transduction of the murine retinal pigmented epithelium (RPE) with adenovirus vectors requires technically difficult and invasive subretinal injections. This study tested the hypothesis that recombinant vectors based on feline immunodeficiency virus (FIV) could access the retina following intravitreal injection. Methods FIV vectors expressing E. coli ,-galactosidase (FIV,gal) were injected alone, or in combination with adenovirus vectors expressing eGFP, into the vitreous of normal mice and eyes evaluated for transgene expression. In further studies, the utility of FIV-mediated gene transfer to correct lysosomal storage defects in the anterior and posterior chambers of eyes was tested using recombinant FIV vectors expressing ,-glucuronidase. FIV,gluc vectors were injected into ,-glucuronidase-deficient mice, an animal model of mucopolysacharridoses type VII. Results The results of this study show that similar to adenovirus, both corneal endothelium and cells of the iris could be transduced following intravitreal injection of FIV,gal. However, in contrast to adenovirus, intravitreal injection of FIV,gal also resulted in transduction of the RPE. Immunohistochemistry following an intravitreal injection of an AdeGFP (adenovirus expressing green fluorescent protein) and FIV,gal mixture confirmed that both viruses mediated transduction of corneal endothelium and cells of the iris, while only FIV,gal transduced cells in the retina. Using the ,-glucuronidase-deficient mouse, the therapeutic efficacy of intravitreal injection of FIV,gluc (FIV expressing ,-glucuronidase) was tested. Intravitreal injection of FIV,gluc to the eyes of ,-glucuronidase-deficient mice resulted in rapid reduction (within 2,weeks) of the lysosomal storage defect within the RPE, corneal endothelium, and the non-pigmented epithelium of the ciliary process. Transgene expression and correction of the lysosomal storage defect remained for at least 12,weeks, the latest time point tested. Conclusion These studies demonstrate that intravitreal injection of FIV-based vectors can mediate efficient and lasting transduction of cells in the cornea, iris, and retina. Copyright © 2002 John Wiley & Sons, Ltd. [source] | ||||||||||