1. Detection of treatment-resistant infectious HIV after genome-directed antiviral endonuclease therapy.
- Author
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De Silva Feelixge HS, Stone D, Pietz HL, Roychoudhury P, Greninger AL, Schiffer JT, Aubert M, and Jerome KR
- Subjects
- Base Sequence, Cell Line, DNA, Viral genetics, Drug Resistance, Viral, Endonucleases metabolism, Exodeoxyribonucleases metabolism, Exodeoxyribonucleases pharmacology, Gene Products, pol genetics, Gene Products, pol metabolism, HEK293 Cells, HIV Infections drug therapy, HIV Infections genetics, HIV Infections therapy, HIV Protease genetics, HIV Protease metabolism, HIV Reverse Transcriptase genetics, HIV Reverse Transcriptase metabolism, Humans, Molecular Sequence Data, Mutation, Phosphoproteins metabolism, Phosphoproteins pharmacology, Transduction, Genetic, Virus Replication drug effects, Anti-HIV Agents pharmacology, HIV Infections virology, HIV-1 drug effects, HIV-1 genetics, Reverse Transcriptase Inhibitors pharmacology, Zinc Fingers
- Abstract
Incurable chronic viral infections are a major cause of morbidity and mortality worldwide. One potential approach to cure persistent viral infections is via the use of targeted endonucleases. Nevertheless, a potential concern for endonuclease-based antiviral therapies is the emergence of treatment resistance. Here we detect for the first time an endonuclease-resistant infectious virus that is found with high frequency after antiviral endonuclease therapy. While testing the activity of HIV pol-specific zinc finger nucleases (ZFNs) alone or in combination with three prime repair exonuclease 2 (Trex2), we identified a treatment-resistant and infectious mutant virus that was derived from a ZFN-mediated disruption of reverse transcriptase (RT). Although gene disruption of HIV protease, RT and integrase could inhibit viral replication, a chance single amino acid insertion within the thumb domain of RT produced a virus that could actively replicate. The endonuclease-resistant virus could replicate in primary CD4(+) T cells, but remained susceptible to treatment with antiretroviral RT inhibitors. When secondary ZFN-derived mutations were introduced into the mutant virus's RT or integrase domains, replication could be abolished. Our observations suggest that caution should be exercised during endonuclease-based antiviral therapies; however, combination endonuclease therapies may prevent the emergence of resistance., (Copyright © 2015 Elsevier B.V. All rights reserved.)
- Published
- 2016
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