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

• Published in: Nature communications 2024-05, Vol.15 (1), p.4018-4018, Article 4018
• Main Authors: Aubert, Martine, Haick, Anoria K., Strongin, Daniel E., Klouser, Lindsay M., Loprieno, Michelle A., Stensland, Laurence, Santo, Tracy K., Huang, Meei-Li, Hyrien, Ollivier, Stone, Daniel, Jerome, Keith R.
• Format: Article
• Language: English
• Subjects: 38/1; 38/22; 45; 45/22; 45/29; 45/70; 45/77; 631/1647/334/1874/345; 631/326/596/1553; 64/60; 692/4017; 692/699/255/2514; 82/29; 82/80; Animal models; Animals; Deoxyribonucleic acid; Dependovirus - genetics; Disease Models, Animal; DNA; DNA, Viral - genetics; Editing; Female; Gene Editing - methods; Genetic engineering; Genetic modification; Genetic Therapy - methods; Genetic Vectors - genetics; Genome editing; Genomes; Hepatotoxicity; Herpes Genitalis - therapy; Herpes Genitalis - virology; Herpes simplex; Herpes Simplex - genetics; Herpes Simplex - therapy; Herpes Simplex - virology; Herpes viruses; Herpesvirus 1, Human - genetics; Herpesvirus 1, Human - physiology; Humanities and Social Sciences; Humans; Infections; Mice; multidisciplinary; Neurological diseases; Neurons; Recurrent infection; Science; Science (multidisciplinary); Vero Cells; Viral Load; Virus Latency - genetics; Virus Shedding; Viruses
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-47940-y

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. The main challenge for anti-HSV therapy is to target latent virus in ganglionic neurons. Here, the authors report a well-tolerated anti-HSV gene editing approach against HSV which targets latent HSV genomes and leads to reductions of ganglionic viral loads, and viral shedding upon reactivation in mouse models.