Adeno-associated virus type 5: transduction efficiency and cell-type specificity in the primate retina

Andrew J Lotery; Grace S Yang; Robert F Mullins; Stephen R Russell; Michael Schmidt; Edwin M Stone; Jonathan D Lindbloom; John A Chiorini; Robert M Kotin; Beverly L Davidson

doi:10.1089/104303403322542301

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Adeno-associated virus type 5: transduction efficiency and cell-type specificity in the primate retina

Journal article

Peer reviewed

Adeno-associated virus type 5: transduction efficiency and cell-type specificity in the primate retina

Andrew J Lotery, Grace S Yang, Robert F Mullins, Stephen R Russell, Michael Schmidt, Edwin M Stone, Jonathan D Lindbloom, John A Chiorini, Robert M Kotin and Beverly L Davidson

Human gene therapy, Vol.14(17), pp.1663-1671

11/20/2003

DOI: 10.1089/104303403322542301

PMID: 14633408

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Abstract

Gene transfer using adeno-associated viruses (AAVs) has been effective for treating inherited retinal diseases in animal models. Further evaluation in primates must be performed prior to clinical application, however, because of the difference between the retina of the primate and those of other animals. Prior work has shown that AAV2 can transduce rod-photoreceptor and RPE cells in the non-human primate retina and that AAV5 is more efficient at transducing photoreceptor cells than AAV2 in the rodent retina. In this study, we evaluated the efficiency of AAV5 in the non-human primate retina after subretinal injections of the vector to distinct anatomic retinal regions (superior, inferior, nasal, macula, temporal). rAAV5 led to a rapid onset of transgene expression (within 2 weeks), with expression persisting up to 10 months. Postoperative electrophysiology studies showed that global retinal function was preserved following gene transfer. Quantitative analysis of gene transfer demonstrated a maximum transduction efficiency of 22% in the injected areas. Evaluation of cell types using confocal microscopy and cone-specific antibodies revealed that AAV5, expressing reporter genes from the cytomegalovirus (CMV) promoter/enhancer, preferentially transduced rods. No significant differences were found in the regional tropism of AAV5 among the five areas injected despite variation in retinal topography. Immunohistochemical studies revealed that the AAV5 receptor, PDGFR-A, is localized to the outer segments of rods but not cones providing a basis for the observed tropism. Our results support the utility of AAV5 for rod photoreceptor degeneration therapies.

Electroretinography

Immunohistochemistry

Cell Line

Cytomegalovirus - genetics

Dependovirus - genetics

Gene Transfer Techniques

Green Fluorescent Proteins

Promoter Regions, Genetic

Retina - metabolism

Humans

Electrophysiology

Macaca fascicularis

Macaca mulatta

Retina - physiology

Microscopy, Confocal

Animals

Enhancer Elements, Genetic

Time Factors

Receptor, Platelet-Derived Growth Factor alpha - genetics

Photoreceptor Cells - metabolism

Adenoviridae - genetics

Genetic Vectors

Luminescent Proteins - metabolism

Details

Title: Subtitle: Adeno-associated virus type 5: transduction efficiency and cell-type specificity in the primate retina
Creators: Andrew J Lotery - Department of Ophthalmology, Roy J. and Lucille A Carver College of Medicine, Iowa City, IA 52242, USA
Grace S Yang
Robert F Mullins
Stephen R Russell
Michael Schmidt
Edwin M Stone
Jonathan D Lindbloom
John A Chiorini
Robert M Kotin
Beverly L Davidson
Resource Type: Journal article
Publication Details: Human gene therapy, Vol.14(17), pp.1663-1671
DOI: 10.1089/104303403322542301
PMID: 14633408
NLM abbreviation: Hum Gene Ther
ISSN: 1043-0342
eISSN: 1557-7422
Publisher: United States
Grant note: HD33531 / NICHD NIH HHS DK54759 / NIDDK NIH HHS
Language: English
Date published: 11/20/2003
Academic Unit: Iowa Neuroscience Institute; Ophthalmology and Visual Sciences
Record Identifier: 9983980008202771

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