Gene editing for latent herpes simplex virus infection reduces viral load and shedding in vivo

Nat Commun. 2024 May 13;15(1):4018. doi: 10.1038/s41467-024-47940-y.

Abstract

Anti-HSV therapies are only suppressive because they do not eliminate latent HSV present in ganglionic neurons, the source of recurrent disease. We have developed a potentially curative approach against HSV infection, based on gene editing using HSV-specific meganucleases delivered by adeno-associated virus (AAV) vectors. Gene editing performed with two anti-HSV-1 meganucleases delivered by a combination of AAV9, AAV-Dj/8, and AAV-Rh10 can eliminate 90% or more of latent HSV DNA in mouse models of orofacial infection, and up to 97% of latent HSV DNA in mouse models of genital infection. Using a pharmacological approach to reactivate latent HSV-1, we demonstrate that ganglionic viral load reduction leads to a significant decrease of viral shedding in treated female mice. While therapy is well tolerated, in some instances, we observe hepatotoxicity at high doses and subtle histological evidence of neuronal injury without observable neurological signs or deficits. Simplification of the regimen through use of a single serotype (AAV9) delivering single meganuclease targeting a duplicated region of the HSV genome, dose reduction, and use of a neuron-specific promoter each results in improved tolerability while retaining efficacy. These results reinforce the curative potential of gene editing for HSV disease.

MeSH terms

  • Animals
  • DNA, Viral / genetics
  • Dependovirus* / genetics
  • Disease Models, Animal
  • Female
  • Gene Editing* / methods
  • Genetic Therapy / methods
  • Genetic Vectors / genetics
  • Herpes Genitalis / therapy
  • Herpes Genitalis / virology
  • Herpes Simplex* / genetics
  • Herpes Simplex* / therapy
  • Herpes Simplex* / virology
  • Herpesvirus 1, Human* / genetics
  • Herpesvirus 1, Human* / physiology
  • Humans
  • Mice
  • Vero Cells
  • Viral Load*
  • Virus Latency / genetics
  • Virus Shedding*

Substances

  • DNA, Viral