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Local and systemic responses following intravitreous injection of AAV2-encoded modified Volvox channelrhodopsin-1 in a genetically blind rat model

Gene Therapy volume 23, pages 158–166 (2016)Cite this article

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Abstract

We previously designed a modified channelrhodopsin-1 (mVChR1) protein chimera with a broader action than that of Chlamydomonas channelrhodopsin-2 and reported that its transduction into retinal ganglion cells can restore visual function in genetically blind, dystrophic Royal College of Surgeons (RCS) rats, with photostimuli ranging from 486 to 640 nm. In the current study, we sought to investigate the safety and influence of mVChR1 transgene expression. Adeno-associated virus type 2 encoding mVChR1 was administered by intravitreous injection into dystrophic RCS rats. Reverse-transcription PCR was used to monitor virus and transgene dissemination and the results demonstrated that their expression was restricted specifically within the eye tissues, and not in non-target organs. Moreover, examination of the blood, plasma and serum revealed that no excess immunoreactivity was present, as determined using standard clinical hematological parameters. Serum antibodies targeting the recombinant adeno-associated virus (rAAV) capsid increased after the injection; however, no increase in mVChR1 antibody was detected during the observation period. In addition, retinal histological examination showed no signs of inflammation in rAAV-injected rats. In conclusion, our results demonstrate that mVChR1 can be exogenously expressed without harmful immunological reactions in vivo. These findings will aid in studies of AAV gene transfer to restore vision in late-stage retinitis pigmentosa.

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Acknowledgements

This work was partly supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (nos. 25462747, 24390393 and 26670750); the Program for the Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (no. 10-06, NIBIO); Translational Research Network program (no. 3805140342); and the Japanese Retinitis Pigmentosa Society. We express our heartfelt appreciation to Ichiro Hagimori of Narita Animal Science Laboratory, whose enormous support and insightful comments were invaluable during the course of this study. We also express our heartfelt appreciation to Chikako Yamada of Center for Developmental Biology, RIKEN for technical advice.

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Authors and Affiliations

  1. Department of Chemistry and Bioengineering, Laboratory of Visual Neuroscience, Iwate University Graduate School of Engineering, Morioka, Iwate, Japan

    E Sugano, K Tabata, M Takahashi, F Nishiyama, H Shimizu, M Sato & H Tomita

  2. Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan

    M Tamai

  3. Clinical Research, Innovation and Education Center, Tohoku University Hospital, Sendai, Miyagi, Japan

    H Tomita

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Correspondence to H Tomita.

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Sugano, E., Tabata, K., Takahashi, M. et al. Local and systemic responses following intravitreous injection of AAV2-encoded modified Volvox channelrhodopsin-1 in a genetically blind rat model. Gene Ther 23, 158–166 (2016). https://doi.org/10.1038/gt.2015.99

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