A Novel Genetic Pathway of Human Immunodeficiency Virus Type 1 Resistance to Stavudine Mediated by the K65R Mutation

Garcı́a-Lerma, J. Gerardo; MacInnes, Hamish; Bennett, Diane; Reid, Patrick; Nidtha, Soumya; Weinstock, Hillard; Kaplan, Jonathan E.; Heneine, Walid

doi:10.1128/jvi.77.10.5685-5693.2003

Cited by 78 publications

(52 citation statements)

References 43 publications

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“…Although combination treatment is highly effective, persistent or suboptimal use of NRTIs can result in the development of reverse transcriptase (RT) mutations that confer drug resistance to this class of inhibitors. Tenofovir (TFV ), abacavir (ABC), zalcitabine (ddC) didanosine (ddI) or stavudine (d4T) can select for the lysine-to-arginine substitution at residue 65 (K65R; Garcia-Lerma et al, 2003;Gu et al, 1994b;Tisdale et al, 1997;Wainberg et al, 1999;Winters et al, 1997). K65R viruses show measurable reduced susceptibility (increase in median inhibiton concentration compared with wild-type) to these drugs and cross-resistance to lamivudine (3TC) and emtricitabine (FTC), but remain fully susceptible to zidovudine (AZT) in vitro (Garcia-Lerma et al, 2003;Harrigan et al, 2000;Tisdale et al, 1997;Wainberg et al, 1999).…”

confidence: 99%

“…Tenofovir (TFV ), abacavir (ABC), zalcitabine (ddC) didanosine (ddI) or stavudine (d4T) can select for the lysine-to-arginine substitution at residue 65 (K65R; Garcia-Lerma et al, 2003;Gu et al, 1994b;Tisdale et al, 1997;Wainberg et al, 1999;Winters et al, 1997). K65R viruses show measurable reduced susceptibility (increase in median inhibiton concentration compared with wild-type) to these drugs and cross-resistance to lamivudine (3TC) and emtricitabine (FTC), but remain fully susceptible to zidovudine (AZT) in vitro (Garcia-Lerma et al, 2003;Harrigan et al, 2000;Tisdale et al, 1997;Wainberg et al, 1999). 3TC, FTC and ABC select for the methionine-to-valine substitution at residue 184 (M184V) in cell culture and in vivo Kavlick et al, 1995;Lanier et al, 2004;Miller et al, 2000;Schinazi et al, 1993;Schuurman et al, 1995;Wainberg et al, 1995).…”

confidence: 99%

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The Balance between NRTI Discrimination and Excision Drives the Susceptibility of HIV-1 RT Mutants K65R, M184V and K65R+M184V

Margot

MacArthur

et al. 2007

Antivir Chem Chemother

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The HIV-1 reverse transcriptase (RT) resistance mutations K65R and M184V occur individually and in combination, and can contribute to decreased treatment responses in patients. In order to understand how these mutations interact with one another to confer drug resistance, the susceptibilities and underlying resistance mechanisms of these mutants to nucleoside RT inhibitors (NRTIs) were determined. Virus carrying K65R have reduced susceptibility to most NRTIs, but retain full susceptibility to zidovudine (AZT). M184V mutants have reduced susceptibility to lamivudine (3TC), emtricitabine (FTC) and didanosine (ddI), and contribute to reduced susceptibility to abacavir; however, they remain fully susceptible to tenofovir (TFV), AZT and stavudine (d4T). In cell culture, the K65R+M184V virus showed slightly increased susceptibility to TFV, AZT and d4T compared with K65R alone, but showed further decreases in susceptibility to 3TC, FTC, ddI and abacavir. There are two major biochemical mechanisms of resistance: altered NRTI binding/incorporation and altered NRTI excision after incorporation. For most NRTIs, the primary mechanism of resistance by K65R, M184V and K65R+M184V mutant RTs is to disrupt the NRTIbinding/incorporation steps. In the case of AZT, however, decreased binding/incorporation by K65R and K65R+M184V was counteracted by decreased AZT excision resulting in wild-type susceptibility. For TFV, decreased excision by K65R and K65R+M184V may partially counteract the K65R-driven decrease in incorporation relative to wild-type resulting in only low levels of TFV resistance. The K65R-mediated effect on decreasing NRTI excision was stronger than for M184V. These studies show that both mechanisms of resistance (binding/incorporation and excision) must be considered when defining resistance mechanisms.

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confidence: 99%

The Balance between NRTI Discrimination and Excision Drives the Susceptibility of HIV-1 RT Mutants K65R, M184V and K65R+M184V

Margot

MacArthur

et al. 2007

Antivir Chem Chemother

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“…We previously found that these isolates are more frequent in recently diagnosed persons than AZTresistant viruses with the T215Y/F mutations (14). Although phenotypically indistinguishable from WT HIV-1, viruses with 215C or 215D acquire the 215Y mutation in vitro more rapidly than WT viruses, likely reflecting the need for only one nucleotide change, compared to two for WT HIV-1 (13,14). Clinical studies also suggest that the presence of the 215D/C substitutions may be associated with an increased risk of virologic failure in antiretroviral-naive adults starting therapy with AZT or d4T (6,33).…”

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confidence: 99%

Transmitted Human Immunodeficiency Virus Type 1 Carrying the D67N or K219Q/E Mutation Evolves Rapidly to Zidovudine Resistance In Vitro and Shows a High Replicative Fitness in the Presence of Zidovudine

Garcı́a-Lerma¹,

MacInnes²,

Bennett³

et al. 2004

J Virol

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Drug-naive patients infected with drug-resistant human immunodeficiency virus type 1 (HIV-1) who initiate antiretroviral therapy show a shorter time to virologic failure than patients infected with wild-type (WT) viruses. Resistance-related HIV genotypes not commonly seen in treated patients, which likely result from reversion or loss of primary resistance mutations, have also been recognized in drug-naive persons. Little work has been done to characterize the patterns of mutations in these viruses and the frequency of occurrence, their association with phenotypic resistance, and their effect on fitness and evolution of resistance. Through the analysis of resistance mutations in 1082 newly diagnosed antiretroviral-naive persons from the United States, we found that 35 of 48 (72.9%) persons infected with HIV-1 containing thymidine analog mutations (TAMs) had viruses that lacked a primary mutation (T215Y/F, K70R, or Q151M). Of these viruses, 9 (25.7%) had only secondary TAMs (D67N, K219Q, M41L, or F77L), and all were found to be sensitive to zidovudine (AZT) and other drugs. To assess the impact of secondary TAMs on the evolution of AZT resistance, we generated recombinant viruses from cloned plasma-derived reverse transcriptase sequences. Two viruses had D67N, three had D67N and K219Q/E, and three were WT. Four site-directed mutants with D67N, K219Q, K219E, and D67N/K219Q were also made in HIV-1 HXB2 . In vitro selection of AZT resistance showed that viruses with D67N and/or K219Q/E acquired AZT resistance mutations more rapidly than WT viruses (36 days compared to 54 days; P ‫؍‬ 0.003). To investigate the factors associated with the rapid selection of AZT mutations in these viruses, we evaluated fitness differences among HXB2 WT and HXB2 D67N or HXB2 D67N/K219Q in the presence of AZT. Both HXB2 D67N/K219Q and HXB2 D67N were more fit than HXB2 WT in the presence of either low or high AZT concentrations, likely reflecting low-level resistance to AZT that is not detectable by phenotypic testing. In the absence of AZT, the fitness cost conferred by D67N or K219Q was modest. Our results demonstrate that viruses with unique patterns of TAMs, including D67N and/or K219Q/E, are commonly found among newly diagnosed persons and illustrate the expanding diversity of revertant viruses in this population. The modest fitness cost conferred by D67N and K219Q supports persistence of these mutants in the untreated population and highlights the potential for secondary transmission. The faster evolution of these mutants toward AZT resistance is consistent with the higher viral fitness in the presence of AZT and shows that these viruses are phenotypically different from WT HIV-1. Our study emphasizes the need for clinical studies to better define the impact of these mutants on treatment responses and evolution of resistance.

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“…This mutation was first identified in vitro by serial passage of HIV-1 in peripheral blood mononuclear cell cultures containing the nucleoside analog RT inhibitor (NRTI) 2Ј,3Ј-dideoxycytidine (ddC) (49) and has subsequently been selected in vitro by many other NRTI (2,9,11,15,16,39,41).…”

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confidence: 99%

The K65R Mutation in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Exhibits Bidirectional Phenotypic Antagonism with Thymidine Analog Mutations

Parikh

Bacheler²,

Koontz

et al. 2006

J Virol

117

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The K65R mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is selected in vitro by many D-nucleoside analog RT inhibitors (NRTI) but has been rarely detected in treated patients. In recent clinical trials, the K65R mutation has emerged frequently in patients experiencing virologic failure on antiretroviral combinations that do not include 3-azidothymidine (AZT). The reason for this change is uncertain. To gain insight, we examined trends in the frequency of K65R in a large genotype database, the association of K65R with thymidine analog mutations (TAMs) and other NRTI mutations, and the viral susceptibility profile of HIV-1 with K65R alone and in combination with TAMs. Among >60,000 clinical samples submitted for genotype analysis that contained one or more NRTI resistance mutations, the frequency of K65R increased from 0.4% in 1998 to 3.6% in 2003. Among samples with K65R, a strong negative association was evident with the TAMs M41L, D67N, L210W, T215Y/F, and K219Q/E (P < 0.005) but not with other NRTI mutations, including the Q151M complex. This suggested that K65R and TAMs are antagonistic. To test this possibility, we generated recombinant HIV-1 encoding K65R in two different TAM backgrounds: M41L/L210W/ T215Y and D67N/K70R/T215F/K219Q. K65R reduced AZT resistance from >50-fold to <2.5-fold in both backgrounds. In addition, TAMs antagonized the phenotypic effect of K65R, reducing resistance to tenofovir, abacavir, 2,3-dideoxycytidine, dideoxyinosine, and stavudine. In conclusion, K65R and TAMs exhibit bidirectional phenotypic antagonism. This antagonism likely explains the negative association of these mutations in genotype databases, the rare emergence of K65R with antiretroviral therapies that contain AZT, and its more frequent emergence with combinations that exclude AZT.

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A Novel Genetic Pathway of Human Immunodeficiency Virus Type 1 Resistance to Stavudine Mediated by the K65R Mutation

Cited by 78 publications

References 43 publications

The Balance between NRTI Discrimination and Excision Drives the Susceptibility of HIV-1 RT Mutants K65R, M184V and K65R+M184V

The Balance between NRTI Discrimination and Excision Drives the Susceptibility of HIV-1 RT Mutants K65R, M184V and K65R+M184V

Transmitted Human Immunodeficiency Virus Type 1 Carrying the D67N or K219Q/E Mutation Evolves Rapidly to Zidovudine Resistance In Vitro and Shows a High Replicative Fitness in the Presence of Zidovudine

The K65R Mutation in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Exhibits Bidirectional Phenotypic Antagonism with Thymidine Analog Mutations

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