Preclinical safety and biodistribution of CRISPR targeting SIV in non-human primates

Tricia H Burdo; Chen Chen; Rafal Kaminski; Ilker K Sariyer; Pietro Mancuso; Martina Donadoni; Mandy D Smith; Rahsan Sariyer; Maurizio Caocci; Shuren Liao; Hong Liu; Wenwen Huo; Huaqing Zhao; John Misamore; Mark G Lewis; Vahan Simonyan; Elaine E Thompson; Ethan Y Xu; Thomas J Cradick; Jennifer Gordon; Kamel Khalili

doi:10.1038/s41434-023-00410-4

Preclinical safety and biodistribution of CRISPR targeting SIV in non-human primates

Gene Ther. 2023 Aug 17. doi: 10.1038/s41434-023-00410-4. Online ahead of print.

Authors

Tricia H Burdo¹, Chen Chen², Rafal Kaminski², Ilker K Sariyer², Pietro Mancuso², Martina Donadoni², Mandy D Smith², Rahsan Sariyer², Maurizio Caocci², Shuren Liao², Hong Liu², Wenwen Huo³, Huaqing Zhao⁴, John Misamore⁵, Mark G Lewis⁵, Vahan Simonyan⁶, Elaine E Thompson⁶, Ethan Y Xu³, Thomas J Cradick³, Jennifer Gordon⁷, Kamel Khalili⁸

Affiliations

¹ Department of Microbiology, Immunology, and Inflammation, Center for NeuroVirology and Gene Editing, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, 19140, USA. burdot@temple.edu.
² Department of Microbiology, Immunology, and Inflammation, Center for NeuroVirology and Gene Editing, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, 19140, USA.
³ Excision BioTherapeutics, Inc., San Francisco, CA, USA.
⁴ Center for Biostatistics and Epidemiology, Department of Biomedical Education and Data Science, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, 19140, USA.
⁵ BioQual, Inc., Rockville, MD, USA.
⁶ Embleema, Metuchen, NJ, USA.
⁷ Excision BioTherapeutics, Inc., San Francisco, CA, USA. jgordon@excisionbio.com.
⁸ Department of Microbiology, Immunology, and Inflammation, Center for NeuroVirology and Gene Editing, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, 19140, USA. kkhalili@temple.edu.

PMID: 37587230
DOI: 10.1038/s41434-023-00410-4

Abstract

In this study, we demonstrate the safety and utility of CRISPR-Cas9 gene editing technology for in vivo editing of proviral DNA in ART-treated, virally controlled simian immunodeficiency virus (SIV) infected rhesus macaques, an established model for HIV infection. EBT-001 is an AAV9-based vector delivering SaCas9 and dual guide RNAs designed to target multiple regions of the SIV genome: the viral LTRs, and the Gag gene. The results presented here demonstrate that a single IV inoculation of EBT-001 at each of 3 dose levels (1.4 × 10¹², 1.4 × 10¹³ and 1.4 × 10¹⁴ genome copies/kg) resulted in broad and functional biodistribution of AAV9-EBT-001 to known tissue reservoirs of SIV. No off-target effects or abnormal pathology were observed, and animals returned to their normal body weight after receiving EBT-001. Importantly, the macaques that received the 2 highest doses of EBT-001 showed improved absolute lymphocyte counts as compared to antiretroviral-treated controls. Taken together, these results demonstrate safety, biodistribution, and in vivo proviral DNA editing following IV administration of EBT-001, supporting the further development of CRISPR-based gene editing as a potential therapeutic approach for HIV in humans.