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PLoS One. 2017 Aug 1;12(8):e0182473. doi: 10.1371/journal.pone.0182473. eCollection 2017.

Single stranded adeno-associated virus achieves efficient gene transfer to anterior segment in the mouse eye.

Author information

1
Department of Ophthalmology and Visual Science and Eye Research Institute, Eye, and ENT Hospital, Shanghai Medical College, Fudan University, Shanghai, China.
2
Key Laboratory of Myopia, Ministry of Health, Fudan University, Shanghai, China.
3
Shanghai Key Laboratory of Visual Impairment and Restoration, Fudan University, Shanghai, China.
4
Grousbeck Gene Therapy Center, Schepens Eye Research Institute and Massachusetts Eye and Ear Infirmary, Boston, Massachusetts, United States of America.
5
Ocular Genomics Institute, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States of America.
6
Harvard Stem Cell Institute, Harvard University, Cambridge, Massachusetts, United States of America.

Abstract

Adeno-associated viruses (AAVs) are used extensively as a gene delivery vehicle for retinal gene therapy, yet its ability to target the anterior segment of the eye, critical to unlocking therapeutic opportunities, is less characterized. Previously, self-complimentary (sc) AAV was shown to be necessary for transduction of the cornea and trabecular meshwork (TM), limiting the size of the gene transfer cassette, likely due to a block in second strand synthesis thought to be required for functional transduction. Here, we evaluated several AAV capsids in a single stranded (ss) genome conformation for their ability to overcome the need for scAAV for targeting corneal endothelium and TM. AAV2, 8, and a recently synthetically developed AAV called Anc80L65 were evaluated in vitro and in vivo by intracameral injection in mice. Results show that although scAAV2 demonstrated superior infectivity in vitro including Human Trabecular meshwork (HTM) immortalized cell lines; Anc80L65 transduced following a single intracameral injection efficiently all components of the mouse anterior segment, including the TM, corneal stroma, and endothelial cells. These results suggest that Anc80L65 is able to overcome the requirement for scAAV genomes to enable TM and corneal targeting, expanding the potential experimental and therapeutic use of AAV gene transfer in the anterior segment of the eye.

PMID:
28763501
PMCID:
PMC5538712
DOI:
10.1371/journal.pone.0182473
[Indexed for MEDLINE]
Free PMC Article

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