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Tyrosine capsid-mutant AAV vectors for gene delivery to the canine retina from a subretinal or intravitreal approach

Gene Therapy volume 21, pages 96–105 (2014)Cite this article

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Abstract

Recombinant adeno-associated viruses are important vectors for retinal gene delivery. Currently utilized vectors have relatively slow onset, and for efficient transduction it is necessary to deliver treatment subretinally, with the potential for damage to the retina. Amino-acid substitutions in the viral capsid improve efficiency in rodent eyes by evading host responses. As dogs are important large animal models for human retinitis pigmentosa, we evaluated the speed and efficiency of retinal transduction using capsid-mutant vectors injected both subretinally and intravitreally. We evaluated AAV serotypes 2 and 8 with amino-acid substitutions of surface-exposed capsid tyrosine residues. The chicken beta-actin promoter was used to drive green fluorescent protein expression. Twelve normal adult beagles were injected; four dogs received intravitreal injections and eight dogs received subretinal injections. Capsid-mutant viruses tested included AAV2(quad Y-F) (intravitreal and subretinal) and self-complementary scAAV8(Y733F) (subretinal only). Contralateral control eyes received injections of scAAV5 (subretinal) or scAAV2 (intravitreal). Subretinally delivered vectors had a faster expression onset than intravitreally delivered vectors. Subretinally delivered scAAV8(Y733F) had a faster onset of expression than scAAV5. All subretinally injected vector types transduced the outer retina with high efficiency and the inner retina with moderate efficiency. Intravitreally delivered AAV2(quad Y-F) had a marginally higher efficiency of transduction of both outer retinal and inner retinal cells than scAAV2. Because of their rapid expression onset and efficient transduction, subretinally delivered capsid-mutant AAV8 vectors may increase the efficacy of gene therapy treatment for rapid photoreceptor degenerative diseases. With further refinement, capsid-mutant AAV2 vectors show promise for retinal gene delivery from an intravitreal approach.

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Acknowledgements

We thank Janice Querubin and Lisa Allen for animal assistance, Joe Hauptman for statistical assistance, Cheryl Craft for hCAR antibody and Vince Chiodo for AAV purification. JTB and FMM acknowledge the following funding sources: Michigan State University College of Veterinary Medicine Endowed Research Fund. SPJ acknowledges the following funding sources: The Glassen Memorial Foundation, Myers-Dunlap Endowment for Canine Health. WWH acknowledges the following funding sources: NIH grant EY021721 and grants from the Macula Vision Research Foundation, Foundation Fighting Blindness, Eldon Family Foundation, Overstreet Fund and Research to Prevent Blindness, Inc.

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Authors and Affiliations

  1. Department of Small Animal Clinical Sciences, Michigan State University, East Lansing, MI, USA

    F M Mowat, K R Gornik, S M Petersen-Jones & J T Bartoe

  2. Department of Ophthalmology, University of Florida College of Medicine, Gainesville, FL, USA

    A Dinculescu, S L Boye & W W Hauswirth

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  1. F M Mowat
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  2. K R Gornik
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  3. A Dinculescu
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  4. S L Boye
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Correspondence to S M Petersen-Jones.

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Competing interests

WWH and the University of Florida have a financial interest in the use of AAV therapies and own equity in a company (AGTC Inc.) that might, in the future, commercialize some aspects of this work. SLB and WWH hold a patent (No. 8 298 818) that covers some aspects of the AAV technology used in this study. The remaining authors declare no conflict of interest.

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Mowat, F., Gornik, K., Dinculescu, A. et al. Tyrosine capsid-mutant AAV vectors for gene delivery to the canine retina from a subretinal or intravitreal approach. Gene Ther 21, 96–105 (2014). https://doi.org/10.1038/gt.2013.64

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