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

Nat Commun. July 2, 2019 V.10 N.1 P.2753.

Dash PK1, Kaminski R2, Bella R2, Su H1, Mathews S1, Ahooyi TM2, Chen C2, Mancuso P2, Sariyer R2, Ferrante P2, Donadoni M2, Robinson JA2, Sillman B1, Lin Z1, Hilaire JR1, Banoub M1, Elango M1, Gautam N3, Mosley RL1, Poluektova LY1, McMillan J1, Bade AN1, Gorantla S1, Sariyer IK2, Burdo TH2, Young WB2, Amini S2, Gordon J2, Jacobson JM2, Edagwa B1, Khalili K4, Gendelman HE5.

Author information

1 Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, 68198-5880, USA.

2 Department of Neuroscience, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, 19115, USA.

3 Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE, 68198-5880, USA.

4 Department of Neuroscience, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, 19115, USA. kamel.khalili@temple.edu.

5 Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, 68198-5880, USA. hegendel@unmc.edu.

Abstract

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.

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6606613/pdf/41467_2019_Article_10366.pdf