CRISPR-Cas9-mediated genome editing delivered by a single AAV9 vector inhibits HSV-1 reactivation in a latent rabbit keratitis model.

Autor:	Amrani N; Excision BioTherapeutics Inc, Watertown, MA, USA., Luk K; Excision BioTherapeutics Inc, Watertown, MA, USA., Singh P; Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI, USA., Shipley M; Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI, USA., Isik M; Excision BioTherapeutics Inc, Watertown, MA, USA., Donadoni M; Center for Neurovirology and Gene Editing, Department of Microbiology, Immunology and Inflammation, Temple University Lewis Katz School of Medicine, Philadelphia, PA, USA., Bellizzi A; Center for Neurovirology and Gene Editing, Department of Microbiology, Immunology and Inflammation, Temple University Lewis Katz School of Medicine, Philadelphia, PA, USA., Khalili K; Center for Neurovirology and Gene Editing, Department of Microbiology, Immunology and Inflammation, Temple University Lewis Katz School of Medicine, Philadelphia, PA, USA., Sariyer IK; Center for Neurovirology and Gene Editing, Department of Microbiology, Immunology and Inflammation, Temple University Lewis Katz School of Medicine, Philadelphia, PA, USA., Neumann D; Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI, USA., Gordon J; Excision BioTherapeutics Inc, Watertown, MA, USA., Ruan GX; Excision BioTherapeutics Inc, Watertown, MA, USA.
Jazyk:	angličtina
Zdroj:	Molecular therapy. Methods & clinical development [Mol Ther Methods Clin Dev] 2024 Aug 14; Vol. 32 (3), pp. 101303. Date of Electronic Publication: 2024 Aug 14 (Print Publication: 2024).
DOI:	10.1016/j.omtm.2024.101303
Abstrakt:	Herpes simples virus 1 (HSV-1) keratitis is a major cause of blindness globally. During primary infection, HSV-1 travels to the trigeminal ganglia and establishes lifelong latency. Although some treatments can reduce symptom severity and recurrence, there is no cure for HSV-1 keratitis. We used CRISPR-Cas9 to co-target gene sequences encoding two essential HSV-1 proteins, ICP0 and ICP27, as a potential therapy for HSV-1 keratitis. In HSV-1-infected Vero cells, the HSV-1 viral load and titer were significantly reduced by plasmid transfection or AAV2 vector transduction expressing Cas9 nuclease from Staphylococcus aureus (SaCas9) and paired guide RNAs (gRNAs). Off-target assessment showed minimal off-target editing activity from the selected gRNAs. We then tested our CRISPR-Cas9 gene editing approach in a latent rabbit model of HSV-1 keratitis. Corneal scarification with all-in-one AAV8(Y733F)-SaCas9 or AAV9-SaCas9 vector reduced viral shedding by over 50%. Interestingly, intravenous administration of the same AAV9-SaCas9 vector eliminated viral shedding in 92% of treated eyes. In addition, treated trigeminal ganglia showed a reduction in HSV-1 DNA and RNA expression. Our results support the utility of single-dose AAV9 all-in-one CRISPR-Cas9 gene editing as a safe and effective strategy for treating HSV-1 keratitis. Competing Interests: N.A., K.L., M.I., J.G., and G.-X.R. are employees of Excision BioTherapeutics. K.K. is a co-founder, board member, scientific advisor, and holds equity in Excision BioTherapeutics. The authors have filed a patent application related to this work. (© 2024 The Authors.)
Databáze:	MEDLINE
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