Biological Sciences, School of

 

School of Biological Sciences: Faculty Publications

Unique Molecular Signatures in Rebound Viruses from Antiretroviral Drug and CRISPR-Treated HIV-1-Infected Humanized Mice

ORCID IDs

Zhang https://orcid.org/0000-0002-0785-1292

Poluektova https://orcid.org/0000-0001-7339-8732

Gendelman https://orcid.org/0000-0002-7831-0370

Dash https://orcid.org/0000-0002-8250-2359

Document Type

Article

Date of this Version

2025

Citation

Communications Biology (2025) 8: 1077

doi: 10.1038/s42003-025-08499-6

Comments

Open access

License: CC BY-NC-ND 4.0

Abstract

HIV-1 elimination from a subset of virus-infected humanized mice (hu-mice) is reported following sequential dual treatment with long-acting (LA) antiretroviral (ART) and CRISPR-Cas9 therapies. However, viral rebound is observed in > 50% of the dual-treated animals. The molecular signatures of the rebound virus, recovered from plasma, have now been determined. The LA-ART treatment contains nanoformulated dolutegravir, lamivudine, abacavir, and rilpivirine combinations, and the HIV-1 excision treatment is CRISPR-Cas9 targeting the HIV-1-LTR-gag. One-step reverse transcriptase polymerase chain reaction, which avoids spontaneous preparatory mutations, is performed on plasma-derived RNA. Sanger and Next-Generation Sequencing are employed targeting the HIV-1-gagpol, and env genes. HIV-1 env shows the most divergence. LA-ART, with or without CRISPR, is responsible for the new mutations. The primary and accessory mutations are detected by deep sequencing. Viral evolution reflects changes in the virus as reported by ART-treated and HIV-1-infected patients. No major CRISPR-specific mutations are observed. The molecular viral signatures demonstrate an accelerated HIV-1 drug resistance escape from ART rather than from the generation of CRISPR mutants. These define viral rebound in the dual-treated hu-mice. The data underscores the limited role of CRISPR excision in generating these rebound HIV-1 mutants from dual-treated hu-mice.

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