Gene editing and elimination of latent herpes simplex virus in vivo

• Published in: Nature communications 2020-08, Vol.11 (1), p.4148-15, Article 4148
• Main Authors: Aubert, Martine, Strongin, Daniel E., Roychoudhury, Pavitra, Loprieno, Michelle A., Haick, Anoria K., Klouser, Lindsay M., Stensland, Laurence, Huang, Meei-Li, Makhsous, Negar, Tait, Alexander, De Silva Feelixge, Harshana S., Galetto, Roman, Duchateau, Philippe, Greninger, Alexander L., Stone, Daniel, Jerome, Keith R.
• Format: Article
• Language: English
• Subjects: 13/106; 38; 38/22; 38/23; 38/70; 38/77; 38/90; 42/41; 631/1647/1511; 631/326/596/1296; 631/326/596/1553; 631/326/596/2564; 64/60; Activation; Animals; Antiviral agents; Cells, Cultured; Chlorocebus aethiops; CRISPR; CRISPR-Cas Systems - genetics; Deoxyribonucleases - genetics; Dependovirus - genetics; Editing; Eye Infections - genetics; Eye Infections - therapy; Eye Infections - virology; Female; Ganglia; Gene Editing - methods; Gene sequencing; Genetic engineering; Genetic modification; Genome editing; HEK293 Cells; Herpes simplex; Herpes Simplex - genetics; Herpes Simplex - therapy; Herpes viruses; Herpesvirus 1, Human - genetics; Herpesvirus 1, Human - pathogenicity; Humanities and Social Sciences; Humans; Infections; Latency; Mice; multidisciplinary; Neurons - metabolism; Neurons - virology; Optimization; Recurrent infection; Ribonucleic acid; RNA; RNA viruses; RNA-Seq; Science; Science (multidisciplinary); Serotypes; Single-Cell Analysis; Superior Cervical Ganglion - metabolism; Superior Cervical Ganglion - virology; Vero Cells; Virus Latency - genetics; Viruses
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-17936-5

We evaluate gene editing of HSV in a well-established mouse model, using adeno-associated virus (AAV)-delivered meganucleases, as a potentially curative approach to treat latent HSV infection. Here we show that AAV-delivered meganucleases, but not CRISPR/Cas9, mediate highly efficient gene editing of HSV, eliminating over 90% of latent virus from superior cervical ganglia. Single-cell RNA sequencing demonstrates that both HSV and individual AAV serotypes are non-randomly distributed among neuronal subsets in ganglia, implying that improved delivery to all neuronal subsets may lead to even more complete elimination of HSV. As predicted, delivery of meganucleases using a triple AAV serotype combination results in the greatest decrease in ganglionic HSV loads. The levels of HSV elimination observed in these studies, if translated to humans, would likely significantly reduce HSV reactivation, shedding, and lesions. Further optimization of meganuclease delivery and activity is likely possible, and may offer a pathway to a cure for HSV infection. Herpes simplex virus establishes lifelong latency in ganglionic neurons, which are the source for recurrent infection. Here Aubert et al. report a promising antiviral therapy based on gene editing with adeno-associated virus-delivered meganucleases, which leads to a significant reduction in ganglionic HSV loads and HSV reactivation.