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Reduced retinal transduction and enhanced transgene-directed immunogenicity with intravitreal delivery of rAAV following posterior vitrectomy in dogs

Gene Therapy volume 23, pages 548–556 (2016)Cite this article

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Abstract

Adeno-associated virus (AAV) vector-based gene therapy is a promising treatment strategy for delivery of neurotrophic transgenes to retinal ganglion cells (RGCs) in glaucoma patients. Retinal distribution of transgene expression following intravitreal injection (IVT) of AAV is variable in animal models and the vitreous humor may represent a barrier to initial vector penetration. The primary goal of our study was to investigate the effect of prior core vitrectomy with posterior hyaloid membrane peeling on pattern and efficiency of transduction of a capsid amino acid substituted AAV2 vector, carrying the green fluorescent protein (GFP) reporter transgene following IVT in dogs. When progressive intraocular inflammation developed starting 4 weeks post IVT, the study plan was modified to allow detailed characterization of the etiology as a secondary goal. Unexpectedly, surgical vitrectomy was found to significantly limit transduction, whereas in non-vitrectomized eyes transduction efficiency reached upwards to 37.3% of RGC layer cells. The developing retinitis was characterized by mononuclear cell infiltrates resulting from a delayed-type hypersensitivity reaction, which we suspect was directed at the GFP transgene. Our results, in a canine large animal model, support caution when considering surgical vitrectomy before IVT for retinal gene therapy in patients, as prior vitrectomy appears to significantly reduce transduction efficiency and may predispose the patient to development of vector-induced immune reactions.

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Acknowledgements

We thank Janice Querbin and Kristin Koehl for animal care assistance, Laurence Occelli for imaging assistance and the laboratory of Lorraine Sordillo for PBMC collection. We also thank Cheryl Craft for hCAR antibody production, Jingfen Sun for performing NAb assays as well as Vince Chiodo and the Retinal Gene Therapy Vector lab for AAV purification. JTB and RFB acknowledge the following funding source: American College of Veterinary Ophthalmologists Vision for Animals Foundation. SLB, SEB, WWH and AMK acknowledge the following funding source: Foundation Fighting Blindness and an unrestricted grant to the UF Department of Ophthalmology from Research to Prevent Blindness. SEB acknowledges the following funding source: NIH Grant R01 EY024280. SEB, SLB and WWH acknowledge funding support from NIH Grant P30EY021721. AMK acknowledges the following funding source: NIH Grant R01 EY019304. SMP-J acknowledges the following funding source: Myers-Dunlap Endowment for Canine Health. WWH acknowledges the following funding sources: NIH grants R01 EY017549, R24 EY022023, and R24 EY022012.

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Author notes

  1. I M Langohr

    Present address: 5Current address: Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USA.,

Authors and Affiliations

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

    R F Boyd, A M Komáromy, S M Petersen-Jones & J T Bartoe

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

    S L Boye, W W Hauswirth & S E Boye

  3. Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA

    T J Conlon & K E Erger

  4. Diagnostic Center for Population and Animal Health, Michigan State University, East Lansing, MI, USA

    D G Sledge & I M Langohr

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Corresponding author

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. The remaining authors declare no conflict of interest.

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Boyd, R., Boye, S., Conlon, T. et al. Reduced retinal transduction and enhanced transgene-directed immunogenicity with intravitreal delivery of rAAV following posterior vitrectomy in dogs. Gene Ther 23, 548–556 (2016). https://doi.org/10.1038/gt.2016.31

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