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Hum Gene Ther. 2019 Aug 28. doi: 10.1089/hum.2019.159. [Epub ahead of print]

Helper-Dependent Adenovirus Transduces the Human and Rat Retina but Elicits an Inflammatory Reaction When Delivered Subretinally in Rats.

Author information

1
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; ian-han@uiowa.edu.
2
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; erin-burnight@uiowa.edu.
3
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; mallory-ulferts@uiowa.edu.
4
University of Iowa, 4083, Institute for Vision Research, Department of Biomedical Engineering, Iowa City, Iowa, United States; kristan-worthington@uiowa.edu.
5
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; stephen-russell@uiowa.edu.
6
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; elliott-sohn@uiowa.edu.
7
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; robert-mullins@uiowa.edu.
8
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; edwin-stone@uiowa.edu.
9
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, 375 Newton Rd, Iowa City, Iowa, United States, 52242; budd-tucker@uiowa.edu.
10
University of Iowa, 4083, Institute for Vision Research, Department of Ophthalmology & Visual Sciences, Iowa City, Iowa, United States; luke-wiley@uiowa.edu.

Abstract

The identification of over 100 genes causing inherited retinal degeneration and the promising results of recent gene augmentation trials have led to an increase in the number of studies investigating the preclinical efficacy of viral-mediated gene transfer. Despite success using adeno-associated viruses (AAV), many disease-causing genes, like ABCA4 or USH2A, are too large to be packaged into these vectors. One option for large gene delivery is the family of integration deficient helper-dependent adenoviruses (HDAds), which efficiently transduce post-mitotic neurons. However, HDAds have been shown in other organ systems to elicit an immune response, and the immunogenicity of HDAds in the retina has not been characterized. In this study, HDAd5 was found to successfully transduce rod and cone photoreceptors in ex vivo human retinal organ cultures. The ocular inflammatory response to subretinal injection of the HDAd serotype 5 (HDAd5) was evaluated using a rat model. Subretinal injection of HDAd5 carrying cytomegalovirus promoter-driven enhanced Green Fluorescent Protein (HDAd5-CMVp-eGFP) elicited a robust inflammatory response by 3-days post-injection. This reaction included vitreous infiltration of Iba1-positive monocytes and increased expression of the pro-inflammatory protein, ICAM-1. By 7-days post-injection, most Iba1-positive infiltrates migrated into the neural retina, including the outer nuclear layer, and ICAM-1 expression was significantly increased compared to buffer-injected contralateral control eyes. At 14-days post-injection, Iba1-positive cells persisted in the retinas of HDAd5-injected eyes, and there was thinning of the outer nuclear layer indicative of photoreceptor cell loss. Subretinal injection of an empty HDAd5 virus was used to confirm that the inflammatory response was in response to the HDAd5 vector and not due to eGFP-induced overexpression cytotoxicity. Despite their larger carrying capacity, further work is needed to elucidate the inflammatory pathways involved and to identify an immunomodulation paradigm sufficient for safe and effective transfer of large genes to the retina using the HDAd5 system.

PMID:
31456426
DOI:
10.1089/hum.2019.159

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