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(Investigative Ophthalmology and Visual Science. 2004;45:3721-3726.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-0059
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Photoreceptor Protection by Iris Pigment Epithelial Transplantation Transduced with AAV-Mediated Brain-Derived Neurotrophic Factor Gene

Masayoshi Hojo,1 Toshiaki Abe,2 Eriko Sugano,1 Yuki Yoshioka,1 Yoko Saigo,1 Hiroshi Tomita,1 Ryosuke Wakusawa,1 and Makoto Tamai1

1From the Department of Ophthalmology and Visual Science and the 2Division of Clinical Cell Therapy, School of Medicine, Tohoku University, Miyagi, Japan.

PURPOSE. To determine whether subretinal transplantation of iris pigment epithelial (IPE) cells transduced with the adenoassociated virus (AAV2)–mediated brain-derived neurotrophic factor (BDNF) gene can protect photoreceptors against phototoxicity.

METHODS. The BDNF gene was inserted into AAV2 (AAV2-BDNF), and the recombinant AAV2 was transduced into rat IPE (AAV2-BDNF-IPE) cells at various multiplicities of infection (MOI). The concentrations of AAV capsids and BDNF were determined by enzyme-linked immunosorbent assay (ELISA). The AAV2-BDNF-IPE cells were transplanted into the subretinal space of rats, and the rats were placed under constant light on days 1 and 90 after the transplantation. The thickness of the outer nuclear layer was measured in histologic sections and compared to that of control sections. The expression of ß-galactosidase (LacZ) in the subretinal space was confirmed by LacZ staining after AAV2-LacZ-IPE transplantation. BDNF gene expression after transplantation was confirmed by real-time polymerase chain reaction (PCR).

RESULTS. Transduction efficiency increased with successive days in culture and increased with higher MOI in vitro. The expression of the BDNF gene in the subretinal space was higher in AAV-BDNF-IPE than with AAV2-LacZ-IPE or with IPE-only transplantation. LacZ expression was observed in the subretinal space 7 and 90 days after transplantation. A statistically significant photoreceptor protection was observed on days 1 and 90 in eyes receiving the AAV2-BDNF-IPE transplant, in both the superior transplant site and the inferior hemispheres which did not receive the transplant.

CONCLUSIONS. Transplantation of AAV2-BDNF-IPE cells may be an alternative method of delivering neurotrophic factors to the lesion.




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