Lethal Mutagenesis of HIV with Mutagenic Nucleoside Analogs

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 96; no. 4; pp. 1492 - 1497
• Main Authors: Loeb, Lawrence A., Essigmann, John M., Kazazi, Farhad, Zhang, Jue, Rose, Karl D., Mullins, James I.
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 16.02.1999; National Acad Sciences; National Academy of Sciences; The National Academy of Sciences
• Subjects: Antimetabolites - pharmacokinetics; Antimetabolites - pharmacology; Antiviral Agents - pharmacokinetics; Antiviral Agents - pharmacology; Biological Sciences; Cell Line; Cell Survival - drug effects; DNA; Genetic mutation; Genetics; Genomes; HIV; HIV Reverse Transcriptase - metabolism; HIV-1 - drug effects; HIV-1 - genetics; Human immunodeficiency virus; Humans; Hybridity; Mutagenesis; Mutation; Nucleic Acid Hybridization; Nucleosides; Nucleosides - pharmacokinetics; Nucleosides - pharmacology; Nucleotides; RNA, Viral - drug effects; RNA, Viral - genetics; Viral DNA; Viruses
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.96.4.1492

The human immunodeficiency virus (HIV) replicates its genome and mutates at exceptionally high rates. As a result, the virus is able to evade immunological and chemical antiviral agents. We tested the hypothesis that a further increase in the mutation rate by promutagenic nucleoside analogs would abolish viral replication. We evaluated deoxynucleoside analogs for lack of toxicity to human cells, incorporation by HIV reverse transcriptase, resistance to repair when incorporated into the DNA strand of an$\text{RNA}· \text{DNA}$hybrid, and mispairing at high frequency. Among the candidates tested, 5-hydroxydeoxycytidine (5-OH-dC) fulfilled these criteria. In seven of nine experiments, the presence of this analog resulted in the loss of viral replicative potential after 9-24 sequential passages of HIV in human CEM cells. In contrast, loss of viral replication was not observed in 28 control cultures passaged in the absence of the nucleoside analog, nor with other analogs tested. Sequence analysis of a portion of the HIV reverse transcriptase gene demonstrated a disproportionate increase in G → A substitutions, mutations predicted to result from misincorporation of 5-OH-dC into the cDNA during reverse transcription. Thus, "lethal mutagenesis" driven by the class of deoxynucleoside analogs represented by 5-OH-dC could provide a new approach to treating HIV infections and, potentially, other viral infections.