June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
In Vivo depletion of the Herpes Simplex Virus 1 Latency Associated Transcript intron using AAV-ribozymes inhibits the establishment of neuronal latency.
Author Affiliations & Notes
  • Mason Shipley
    Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Pankaj Singh
    Department of Ophthalmology & Visual Science, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • David Clair Bloom
    Molecular Genetics & Microbiology, University of Florida, Gainesville, Florida, United States
  • Donna M Neumann
    Department of Pathology and Laboratory Medicine, University of Wisconsin-Madison, Madison, Wisconsin, United States
    Department of Ophthalmology & Visual Science, University of Wisconsin-Madison, Madison, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Mason Shipley None; Pankaj Singh None; David Bloom None; Donna Neumann None
  • Footnotes
    Support  NIAID R01 AI134807; NIAID R01 AI048633; NIH 2TL1TR002375-06
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 145. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Mason Shipley, Pankaj Singh, David Clair Bloom, Donna M Neumann; In Vivo depletion of the Herpes Simplex Virus 1 Latency Associated Transcript intron using AAV-ribozymes inhibits the establishment of neuronal latency.. Invest. Ophthalmol. Vis. Sci. 2023;64(8):145.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Herpes Simplex Virus-1 (HSV-1) is a leading cause of infectious blindness due to repeated reactivation of latent virus from the neuron to the eye. However, the mechanisms that govern the establishment of latency and reactivation are not well characterized. We have shown that a viral long noncoding RNA called the latency associated transcript (LAT) and its intron are required for efficient reactivation. Degradation of these LAT elements results in an attenuated reactivation phenotype. However, the roles that the expression and accumulation of LAT have on the establishment of latency are poorly understood, and previous studies aimed at understanding LAT have only leveraged mutant viruses with large deletions. In this work, we use the topical corneal delivery of Adeno-associated virus (rAAV) vectors containing a LAT intron targeting ribozyme to determine how intron expression dictates the establishment of latency.

Methods : New Zealand White (NZW) rabbits were ocularly administered a rAAV vector containing a ribozyme specific for either the LAT intron (LATRz992) or a Rhodopsin control (N=8). Rabbits were subsequently infected with HSV-1 and maintained for 28 days post-infection, at which time latency is considered to be established. Trigeminal ganglia (TG) were harvested and subjected to either qRT-PCR for gene expression or qPCR to determine viral genome loads. All expression was normalized to the endogenous control GAPDH. Two-tailed Student’s t-test was used for statistical analysis of PCR and Cox regression was used for survival analysis.

Results : Establishment of latency, shown by genomic load in TG, was significantly reduced in rabbits pretreated with LATRz992 (5.44±0.17 Fold change) compared to control (p=0.01) . Overall intron expression was reduced (10.2±0.08 Fold change) when compared to control. LATRz992 treated rabbits had significantly higher mortality rates (p=0.031) following infection with HSV-1 when compared to control, suggesting that lytic replication is occurring in the neuron.

Conclusions : We found that the LAT intron is essential for the establishment of latency in neurons. These data further suggest that future strategies aimed at preventing lifelong HSV-1 latency target the LAT intron during the initial infection.

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

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×