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

Hong Liu, 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.
Chen Chen, 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.
Shuren Liao, 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.
Danielle K. Sohaii, Center for Cancer and Immunology Research, Children's National Health System, The George Washington University, 7144 13th Place NW, Washington, DC 20012, USA.
Conrad R. Cruz, Center for Cancer and Immunology Research, Children's National Health System, The George Washington University, 7144 13th Place NW, Washington, DC 20012, USA.
Tricia H. Burdo, 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.
Thomas J. Cradick, Excision Biotherapeutics, Inc., 499 Jackson Street, San Francisco, CA 94111, USA.
Anand Mehta, Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Basic Science Building, Room 310, 173 Ashley Avenue, Charleston, SC 29425, USA.
Carlos Barrero, Department of Pharmaceutical Sciences, School of Pharmacy, Temple University, 3307 N. Broad Street, Philadelphia, PA 19140, USA.
Magda Florez, Department of Pharmaceutical Sciences, School of Pharmacy, Temple University, 3307 N. Broad Street, Philadelphia, PA 19140, USA.
Jennifer Gordon, Excision Biotherapeutics, Inc., 499 Jackson Street, San Francisco, CA 94111, USA.
Stephane Grauzam, Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Basic Science Building, Room 310, 173 Ashley Avenue, Charleston, SC 29425, USA.
James Dressman, Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Basic Science Building, Room 310, 173 Ashley Avenue, Charleston, SC 29425, USA.
Shohreh Amini, Department of Biology, College of Science and Technology, Temple University, 1900 North 12th Street, Philadelphia, PA 19122, USA.
Catherine M. Bollard, Center for Cancer and Immunology Research, Children's National Health System, The George Washington University, 7144 13th Place NW, Washington, DC 20012, USA.
Rafal Kaminski, 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.
Kamel Khalili, 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.

Document Type

Journal Article

Publication Date

6-13-2023

Journal

Molecular therapy. Nucleic acids

Volume

32

DOI

10.1016/j.omtn.2023.04.027

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

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.

Department

Pediatrics

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