Strategic self-limiting production of infectious HIV particles by CRISPR in permissive cells

Hong Liu; Chen Chen; Shuren Liao; Danielle K Sohaii; Conrad R Y Cruz; Tricia H Burdo; Thomas J Cradick; Anand Mehta; Carlos Barrero; Magda Florez; Jennifer Gordon; Stephane Grauzam; James Dressman; Shohreh Amini; Catherine M Bollard; Rafal Kaminski; Kamel Khalili

doi:10.1016/j.omtn.2023.04.027

Strategic self-limiting production of infectious HIV particles by CRISPR in permissive cells

Mol Ther Nucleic Acids. 2023 May 19:32:1010-1025. doi: 10.1016/j.omtn.2023.04.027. eCollection 2023 Jun 13.

Authors

Hong Liu¹, Chen Chen¹, Shuren Liao¹, Danielle K Sohaii², Conrad R Y Cruz², Tricia H Burdo¹, Thomas J Cradick³, Anand Mehta⁴, Carlos Barrero⁵, Magda Florez⁵, Jennifer Gordon³, Stephane Grauzam⁴, James Dressman⁴, Shohreh Amini⁶, Catherine M Bollard², Rafal Kaminski¹, Kamel Khalili¹

Affiliations

¹ Center for Neurovirology and Gene Editing, Department of Microbiology, Immunology, and Inflammation, Lewis Katz School of Medicine at Temple University, 3500 N. Broad Street, 7th Floor, Philadelphia, PA 19140, USA.
² Center for Cancer and Immunology Research, Children's National Health System, The George Washington University, 7144 13th Place NW, Washington, DC 20012, USA.
³ Excision Biotherapeutics, Inc., 499 Jackson Street, San Francisco, CA 94111, USA.
⁴ Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Basic Science Building, Room 310, 173 Ashley Avenue, Charleston, SC 29425, USA.
⁵ Department of Pharmaceutical Sciences, School of Pharmacy, Temple University, 3307 N. Broad Street, Philadelphia, PA 19140, USA.
⁶ Department of Biology, College of Science and Technology, Temple University, 1900 North 12th Street, Philadelphia, PA 19122, USA.

Abstract

Post-translational glycosylation of the HIV-1 envelope protein involving precursor glycan trimming by mannosyl oligosaccharide glucosidase (MOGS) is critically important for morphogenesis of virions and viral entry. Strategic editing of the MOGS gene in T lymphocytes and myeloid origin cells harboring latent proviral DNA results in the production of non-infectious particles upon treatment of cells with latency reversal agents. Controlled activation of CRISPR-MOGS by rebound HIV-1 mitigates production of infectious particles that exhibit poor ability of the virus to penetrate uninfected cells. Moreover, exclusive activation of CRISPR in cells infected with HIV-1 alleviates concern for broad off-target impact of MOGS gene ablation in uninfected cells. Combination CRISPR treatment of peripheral blood lymphocytes prepared from blood of people with HIV-1 (PWH) tailored for editing the MOGS gene (CRISPR-MOGS) and proviral HIV-1 DNA (CRISPR-HIV) revealed a cooperative impact of CRISPR treatment in inhibiting the production of infectious HIV-1 particles. Our design for genetic inactivation of MOGS by CRISPR exhibits no detectable off-target effects on host cells or any deleterious impact on cell survival and proliferation. Our findings offer the development of a new combined gene editing-based cure strategy for the diminution of HIV-1 spread after cessation of antiretroviral therapy (ART) and its elimination.

Keywords: CRISPR gene editing; CRISPR prevention of viral entry; HIV infection; MT: RNA/DNA Editing; combined gene editing for HIV cure; glycosylation; mannosyl oligosaccharide glucosidase (MOGS); non-infectious virus production.