June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Corneal Application of rAAV-Delivered Ribozymes that Target the LAT of HSV-1 Significantly Reduce Viral Reactivation in the Eye
Author Affiliations & Notes
  • Donna M Neumann
    Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • David Clair Bloom
    Microbiology and Molecular Genetics, University of Florida, Gainesville, Florida, United States
  • Pankaj Singh
    Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Mason Shipley
    Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Donna Neumann None; David Bloom None; Pankaj Singh None; Mason Shipley None
  • Footnotes
    Support  NIH NIAID R01AI134807; NIH NIAID R01AI048633; NIH NEI P30EY16665
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 3798. doi:
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      Donna M Neumann, David Clair Bloom, Pankaj Singh, Mason Shipley; Corneal Application of rAAV-Delivered Ribozymes that Target the LAT of HSV-1 Significantly Reduce Viral Reactivation in the Eye. Invest. Ophthalmol. Vis. Sci. 2023;64(8):3798.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Herpes Simplex Virus 1 (HSV-1) is a lifelong human pathogen that infects >70% of the population by early adulthood. Following the initial infection, the virus establishes a latent viral reservoir in sensory neurons. Latent virus can reactivate from neurons resulting in recurrent ocular infections. HSV-1 is a leading infectious cause of blindness worldwide, and available antivirals do not target latent virus, where only one transcript is expressed; the lncRNA known as the Latency Associated Transcript (LAT). LAT is spliced into a stable intron, whose expression in neurons is required for efficient reactivation, suggesting that the LAT could be a therapeutic target to prevent recurrences. We hypothesized that degrading the LAT intron in latently infected neurons would attenuate reactivation and viral recurrence in the eye.

Methods : 30 NZW rabbits were infected with HSV-1 following corneal abrasion and allowed to establish latency. Latent rabbits were administered rAAV vectors containing ribozymes targeting the 5’ exon or the intron of LAT, or a combination of both. Control vectors were a rhodopsin ribozyme. rAAVs were topically delivered to the cornea following abrasion and rabbits were then subjected to reactivation (epinephrine). To determine reactivation frequencies, both eyes were swabbed daily for 12 days to quantify infectious virus by plaque assay. Statistical analyses were done by Chi-squared analyses.

Results : We found that the individual treatments with the rAAV-ribozyme targeted to the LAT 5’exon or the LAT intron resulted in significant decreases in the frequency and number of rabbit eyes shedding infectious virus compared to controls (83.3% vs 27.3% and 50%, respectively; n=10-12 eyes; (p>0.0001; p>0.002)). Combination treatment using both the intron and exon targeting ribozymes also resulted in a significant reduction of reactivation frequencies compared to controls (83.3% vs 58.3%, p>0.004, n=10-12) but the increase in infectious virus shedding in the combination treatment from single vector therapies suggests that further optimization of ribozyme doses is required for the combination therapy.

Conclusions : Deegrading either the LAT 5’exon or the LAT intron individually using rAAV-delivered ribozymes significantly reduced HSV-1 reactivation, suggesting that targeting the latent HSV-1 reservoir in neurons can prevent HSV-1 recurrence in the eye.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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