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E. Sugano, H. Tomita, H. Isago, T. Hiroi, Z. Wang, M. Tamai; Systematic and Local Responses of Channelrhodopsin-2 Gene Therapy. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3299.
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© ARVO (1962-2015); The Authors (2016-present)
We previously reported that the transduction of the channelrhodopsin-2 gene (ChR2) into the retinal ganglion cells could restore the visual function in the genetically blind rats. The purpose of this study is to reveal the influence of exogenous expression of ChR2.
Adeno-associated virus(AAV) vector encoding ChR2 gene fused with vinus protein encoding gene (ChR2V) was intravitreously injected into both eyes of aged dystrophic Royal College of Surgeons(RCS)rats. Visually evoked potentials (VEPs) were periodically recorded. Peripheral Blood was collected and analysis of T- Lymphocyts subsets was performed by flow-cytometry. To study the immunological reaction to ChR2, the production of the antibody was investigated. Histological study was performed by HE staining. The function of retinal cells was studied on frozen sections by immunostainigs of GS, GFAP or NFkB p65. As an inflammation model, non-degenerated RCS (+/+) rats received intravitreously injection of LPS.
Maximum amplitude of VEP was recorded 8 weeks after the injection of AAV-ChR2V and the response was kept for the observation period, 64 weeks, without the decay. Analysis of T-Lymphocytes revealed expansion of Tregulatory in the positive control, LPS-injected group, but not in AAV-ChR2 injected group. Histological studies showed that no differences of retinal histology between normal and ChR2-expressed retina though leukocyte migration was caused in LPS-injected group. The production of antibody to ChR2 was detected, but there were quietly low level compared to ChR2 antibody produced by immunized rabbit. GS- and GFAP- like immuno-reactivities were observed in the ILM of the ChR2V-expressed retina On the other hand, in retina of no-treated RCS and AAV-Venus injected RCS, the reactivities were observed in INL and the IPL in addition to the staining of the ILM. NFKB like immuno-reactivities in ChR2V-expressed retina were higher than those in no-treated and AAV-Venus injected RCS rats.
The administration of AAV-ChR2 to the eye appears well tolerated. Histological studies showed that expression of ChR2 had some protective effects on inner retinal neurons. These data demonstrated ChR2 transgene into retina with AAV vector is an applicable method for restoring vision.
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