Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice

• Published in: Nature communications Vol. 10; no. 1; pp. 2753 - 20
• Main Authors: Dash, Prasanta K., Kaminski, Rafal, Bella, Ramona, Su, Hang, Mathews, Saumi, Ahooyi, Taha M., Chen, Chen, Mancuso, Pietro, Sariyer, Rahsan, Ferrante, Pasquale, Donadoni, Martina, Robinson, Jake A., Sillman, Brady, Lin, Zhiyi, Hilaire, James R., Banoub, Mary, Elango, Monalisha, Gautam, Nagsen, Mosley, R. Lee, Poluektova, Larisa Y., McMillan, JoEllyn, Bade, Aditya N., Gorantla, Santhi, Sariyer, Ilker K., Burdo, Tricia H., Young, Won-Bin, Amini, Shohreh, Gordon, Jennifer, Jacobson, Jeffrey M., Edagwa, Benson, Khalili, Kamel, Gendelman, Howard E.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 02.07.2019; Nature Publishing Group; Nature Portfolio
• Subjects: 38/77; 42; 42/109; 631/326/4041/3196; 631/326/596/1296; 692/699/255/1901; Adoptive Transfer; Animals; Anti-HIV Agents - administration & dosage; Antiretroviral drugs; Antiretroviral therapy; Antiviral agents; Bone marrow; Brain; Combined Modality Therapy; CRISPR; CRISPR-Cas Systems; Deoxyribonucleic acid; DNA; DNA, Viral - genetics; DNA, Viral - immunology; Gag protein; Gene Editing; HIV; HIV Infections - drug therapy; HIV Infections - immunology; HIV Infections - therapy; HIV Infections - virology; HIV-1 - genetics; HIV-1 - immunology; HIV-1 - physiology; Human immunodeficiency virus; Humanities and Social Sciences; Humans; In vivo methods and tests; Infectious diseases; Lasers; Lymphoid tissue; Mice; multidisciplinary; Progeny; Science; Science (multidisciplinary); Target detection; Treatment Outcome; Virus Latency; Viruses
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-019-10366-y

Elimination of HIV-1 requires clearance and removal of integrated proviral DNA from infected cells and tissues. Here, sequential long-acting slow-effective release antiviral therapy (LASER ART) and CRISPR-Cas9 demonstrate viral clearance in latent infectious reservoirs in HIV-1 infected humanized mice. HIV-1 subgenomic DNA fragments, spanning the long terminal repeats and the Gag gene, are excised in vivo, resulting in elimination of integrated proviral DNA; virus is not detected in blood, lymphoid tissue, bone marrow and brain by nested and digital-droplet PCR as well as RNAscope tests. No CRISPR-Cas9 mediated off-target effects are detected. Adoptive transfer of human immunocytes from dual treated, virus-free animals to uninfected humanized mice fails to produce infectious progeny virus. In contrast, HIV-1 is readily detected following sole LASER ART or CRISPR-Cas9 treatment. These data provide proof-of-concept that permanent viral elimination is possible. Here, the authors show that sequential treatment with long-acting slow-effective release ART and AAV9- based delivery of CRISPR-Cas9 results in undetectable levels of virus and integrated DNA in a subset of humanized HIV-1 infected mice. This proof-of-concept study suggests that HIV-1 elimination is possible.