April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Expression of CNGB3 provides long-term rescue in the cone-only CNGB3 x NRL double knock-out (DKO) mouse
Author Affiliations & Notes
  • Seok-Hong Min
    Ophthalmology, University of Florida, Gainesville, FL
  • Qing Ruan
    Ophthalmology, University of Florida, Gainesville, FL
  • Sanford L Boye
    Ophthalmology, University of Florida, Gainesville, FL
  • Shannon Boye
    Ophthalmology, University of Florida, Gainesville, FL
  • William W Hauswirth
    Ophthalmology, University of Florida, Gainesville, FL
  • Christine Nichols Kay
    Ophthalmology, University of Florida, Gainesville, FL
  • Footnotes
    Commercial Relationships Seok-Hong Min, None; Qing Ruan, None; Sanford Boye, None; Shannon Boye, None; William Hauswirth, AGTC (C); Christine Kay, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 3309. doi:https://doi.org/
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    • Get Citation

      Seok-Hong Min, Qing Ruan, Sanford L Boye, Shannon Boye, William W Hauswirth, Christine Nichols Kay; Expression of CNGB3 provides long-term rescue in the cone-only CNGB3 x NRL double knock-out (DKO) mouse. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3309. doi: https://doi.org/.

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

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Abstract
 
Purpose
 

Achromatopsia (ACHM) is a congenital cone photoreceptor disorder with an incidence of 1:30,000. Cyclic nucleotide gated channel beta 3 (CNGB3) has been reported to account for approximately 50% of this autosomal recessive disease. CNGB3 KO mice have been used successfully as an animal model for gene therapy, but the translation from a rod-dominant murine model to human cone dystrophies is limited. Here, our goal was to test the effects of CNGB3 gene replacement in a CNGB3 x NRL double knock-out (DKO) mouse which more closely resembles the cone dominant macula of ACHM patients.

 
Methods
 

We generated photoreceptor-specific AAV vectors to express CNGB3 in CNGB3 x NRL DKO mice. One microliter (1 x 1012 vector genomes/ml) of AAV8(Y733F) vectors containing the photoreceptor-specific IRBP/GNAT2 promoter driving a human CNGB3 cDNA was delivered subretinally to one eye of CNGB3 x NRL DKO mice at 2 weeks of age. Contralateral eyes were untreated and served as controls. Retinal function was assessed by cone electroretinography (ERG) under photopic conditions out to 8 months post-injection. Responses from both M- and S- cones were isolated individually. Histological analysis is underway.

 
Results
 

At eight months post-injection, AAV8(Y733F)-CNGB3 treated eyes exhibited significantly increased ERG amplitudes (nearly double) compared to contralateral untreated controls. Further analyses showed that treatment increased isolated responses mediated by either M- or S-cone opsin, with larger improvements in the S-cone opsin mediated response.

 
Conclusions
 

Expression of CNGB3 driven by the IRBP/GNAT2 promoter in an AAV vector led to stable (at least 8 months) and significant increases in cone ERG amplitudes. Given its cone only makeup, the CNGB3 x NRL DKO mouse may provide a useful model for relevant proof of concept testing of gene replacement strategies for human achromatopsia.

  
Keywords: 538 gene transfer/gene therapy • 510 electroretinography: non-clinical  
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