June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Tropisms of AAV for Subretinal Delivery to the Neonatal Mouse Retina and Its Application for In Vivo Rescue of the Crx Knockout Retina
Author Affiliations & Notes
  • Satoshi Watanabe
    Laboratory for Molecular and Developmental Biology, Institute for Protein Research, Osaka University and JST, CREST, Suita, Japan
    Department of Molecular Genetics, Kyoto University Graduate School of Medicine, Kyoto, Japan
  • Rikako Sanuki
    Laboratory for Molecular and Developmental Biology, Institute for Protein Research, Osaka University and JST, CREST, Suita, Japan
  • Shinji Ueno
    Department of Ophthalmology, Nagoya University Graduate of School of Medicine, Nagoya, Japan
  • Takahisa Furukawa
    Laboratory for Molecular and Developmental Biology, Institute for Protein Research, Osaka University and JST, CREST, Suita, Japan
  • Footnotes
    Commercial Relationships Satoshi Watanabe, None; Rikako Sanuki, None; Shinji Ueno, None; Takahisa Furukawa, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2732. doi:
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      Satoshi Watanabe, Rikako Sanuki, Shinji Ueno, Takahisa Furukawa, ; Tropisms of AAV for Subretinal Delivery to the Neonatal Mouse Retina and Its Application for In Vivo Rescue of the Crx Knockout Retina. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2732.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Adeno-associated virus (AAV) is well established as a vehicle for in vivo gene transfer into the mammalian retina. This virus is promising not only for gene therapy of retinal diseases, but also for in vivo functional analysis of retinal genes. Previous reports have shown that AAV can infect various cell types in the developing mouse retina. However, AAV tropism in the developing retina has not yet been examined in detail. Thus, we examined the tropisms of seven AAV serotypes (AAV2/1, 2/2, 2/5, 2/8, 2/9, 2/10, and 2/11) for subretinal delivery into the P0 mouse retina.

Methods: We subretinally delivered seven AAV serotypes of AAV-CAG-mCherry into P0 mouse retinas, and quantitatively evaluated the tropisms of each serotype by its infecting degree in retinal cells. After subretinal injection of AAV into postnatal day 0 (P0) mouse retinas, various retinal cell types were efficiently transduced with different AAVs. To confirm the usefulness of AAV-mediated gene transfer into the P0 mouse retina, we performed AAV-mediated rescue of the Cone-rod homeobox gene knockout (Crx KO) mouse, which exhibits an outer segment formation defect, flat electroretinogram (ERG) responses, and photoreceptor degeneration. We subretinally injected an AAV expressing Crx under the control of the Crx 2kb promoter into the neonatal Crx KO retina.

Results: Photoreceptor cells were efficiently transduced with AAV2/5. Retinal cells, except for bipolar and Müller glial cells, were efficiently transduced with AAV2/9. Horizontal and/or ganglion cells were efficiently transduced with AAV2/1, AAV2/2, AAV2/8, AAV2/9 and AAV2/10. We showed that AAV-mediated Crx expression significantly decreased the abnormalities of the Crx KO retina by histological and physiological analyses.

Conclusions: In the current study, we report suitable AAV tropisms for delivery into the developing mouse retina. Using AAV2/5 in photoreceptor cells, we demonstrated the possibility of gene replacement for the developmental disorder and subsequent degeneration of retinal photoreceptors caused by the absence of Crx.

Keywords: 538 gene transfer/gene therapy • 696 retinal degenerations: hereditary • 739 transcription factors  
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