Effects of Raltegravir or Elvitegravir Resistance Signature Mutations on the Barrier to Dolutegravir Resistance
            <i>In Vitro</i>

Seki, Takahiro; Suyama-Kagitani, Akemi; Kawauchi-Miki, Shinobu; Miki, Shigeru; Wakasa-Morimoto, Chiaki; Akihisa, Erika; Nakahara, Koichiro; Kobayashi, Masanori; Underwood, Mark; Sato, Akihiko; Fujiwara, Tamio; Yoshinaga, Tomokazu

doi:10.1128/aac.04844-14

Cited by 32 publications

(16 citation statements)

References 34 publications

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“…Interestingly, the selection of E92Q/ N155H/R263K in response to EVG was reported in that study (18), and we show here that this virus is highly resistant to this INSTI.…”

Section: Discussionmentioning

confidence: 62%

Dolutegravir-Selected HIV-1 Containing the N155H and R263K Resistance Substitutions Does Not Acquire Additional Compensatory Mutations under Drug Pressure That Lead to Higher-Level Resistance and Increased Replicative Capacity

Anstett

Fusco

Cutillas

et al. 2015

J Virol

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We have previously shown that the addition of the raltegravir/elvitegavir (RAL/EVG) primary resistance mutation N155H to the R263K dolutegravir (DTG) resistance mutation partially compensated for the fitness cost imposed by R263K while also slightly increasing DTG resistance in vitro (K. Anstett, T. Mesplede, M. Oliveira, V. Cutillas, and M. A. Wainberg, J Virol 89:4681-4684, 2015, doi:10.1128/JVI.03485-14). Since many patients failing RAL/EVG are given DTG as part of rescue therapy, and given that the N155H substitution often is found in combination with other compensatory resistance mutations in such individuals, we investigated the effects of multiple such substitutions within integrase (IN) on each of integrase function, HIV-1 infectivity, and levels of drug resistance. To this end, each of the L74M, E92Q, T97A, E157Q, and G163R substitutions were introduced into NL4.3 subtype B HIV-1 vectors harboring N155H and R263K in tandem [termed NL4.3 IN(N155H/R263K) ]. Relevant recombinant integrase enzymes also were expressed, and purified and biochemical assays of strand transfer efficiency as well as viral infectivity and drug resistance studies were performed. We found that the addition of T97A, E157Q, or G163R somewhat improved the affinity of IN N155H/R263K for its target DNA substrate, while the presence of L74M or E92Q had a negative effect on this process. However, viral infectivity was significantly decreased from that of NL4.3 IN(N155H/R263K) after the addition of each tertiary mutation, and no increases in levels of DTG resistance were observed. This work shows that the compensatory mutations that evolve after N155H under continued DTG or RAL/EVG pressure in patients are unable to improve either enzyme efficiency or viral infectivity in an N155H/R263K background. IMPORTANCEIn contrast to other drugs, dolutegravir has not selected for resistance in HIV-positive individuals when used in first-line therapy. We had previously shown that HIV containing the primary raltegravir/elvitegravir resistance substitution N155H could select for R263K under dolutegravir pressure and that this virus was fit and displayed low-level resistance to dolutegravir (Anstett et al., J Virol 89:4681-4684). Therefore, the current study aimed to uncover whether accessory mutations that appear after N155H in response to raltegravir/elvitegravir were compatible with N155H and R263K. We found, however, that the addition of a third mutation negatively impacted both the enzyme and the virus in terms of activity and infectivity without large shifts in integrase inhibitor resistance. Thus, it is unlikely that these substitutions would be selected under dolutegravir pressure. These data support the hypothesis that primary resistance against DTG cannot evolve through RAL/EVG resistance pathways and that the selection of R263K leads HIV into an evolutionary dead-end. Integrase strand transfer inhibitors (INSTIs) constitute the newest class of drugs approved for the treatment of HIV. They act by inhibiting the HIV enzyme integrase (IN) ...

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“…Interestingly, the selection of E92Q/ N155H/R263K in response to EVG was reported in that study (18), and we show here that this virus is highly resistant to this INSTI.…”

Section: Discussionmentioning

confidence: 62%

Dolutegravir-Selected HIV-1 Containing the N155H and R263K Resistance Substitutions Does Not Acquire Additional Compensatory Mutations under Drug Pressure That Lead to Higher-Level Resistance and Increased Replicative Capacity

Anstett

Fusco

Cutillas

et al. 2015

J Virol

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“…While resistance mutations were detected within 29 weeks under either DTG or EVG selection pressure, no resistance mutations emerged under pressure with RAL. A direct comparison cannot be drawn between our selection studies, which were performed using rhesus PBMCs, and those performed by others who used HIV-1 in MT-2 cell lines and showed that mutations emerged rapidly under RAL pressure (31,46). Our selection experiments with RAL may suggest that RAL possesses a higher genetic barrier than the other INSTIs in rhesus macaques; however, a previous study revealed that SHIV-infected macaques treated with pressure from L-870812, a RAL analogue, possessed the N155H substitution after 25 days of treatment with this drug (36).…”

Section: Discussionmentioning

confidence: 70%

Characterization of the Drug Resistance Profiles of Integrase Strand Transfer Inhibitors in Simian Immunodeficiency Virus SIVmac239

Hassounah

Liu

Quashie

et al. 2015

J Virol

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We previously showed that the simian immunodeficiency virus SIVmac239 is susceptible to human immunodeficiency virus (HIV) integrase (IN) strand transfer inhibitors (INSTIs) and that the same IN drug resistance mutations result in similar phenotypes in both viruses. Now we wished to determine whether tissue culture drug selection studies with SIV would yield the same resistance mutations as in HIV. Tissue culture selection experiments were performed using rhesus macaque peripheral blood mononuclear cells (PBMCs) infected with SIVmac239 viruses in the presence of increasing concentrations of dolutegravir (DTG), elvitegravir (EVG), and raltegravir (RAL). We now show that 22 weeks of selection pressure with DTG yielded a mutation at position R263K in SIV, similar to what has been observed in HIV, and that selections with EVG led to emergence of the E92Q substitution, which is a primary INSTI resistance mutation in HIV associated with EVG treatment failure. To study this at a biochemical level, purified recombinant SIVmac239 wild-type (WT) and E92Q, T97A, G118R, Y143R, Q148R, N155H, R263K, E92Q T97A, E92Q Y143R, R263K H51Y, and G140S Q148R recombinant substitution-containing IN enzymes were produced, and each of the characteristics strand transfer, 3=-processing activity, and INSTI inhibitory constants was assessed in cell-free assays. The results show that the G118R and G140S Q148R substitutions decreased K m = and V max =/K m = for strand transfer compared to those of the WT. RAL and EVG showed reduced activity against both viruses and against enzymes containing Q148R, E92Q Y143R, and G140S Q148R. Both viruses and enzymes containing Q148R and G140S Q148R showed moderate levels of resistance against DTG. This study further confirms that the same mutations associated with drug resistance in HIV display similar profiles in SIV. IMPORTANCEOur goal was to definitively establish whether HIV and simian immunodeficiency virus (SIV) share similar resistance pathways under tissue culture drug selection pressure with integrase strand transfer inhibitors and to test the effect of HIV-1 integrase resistance-associated mutations on SIV integrase catalytic activity and resistance to integrase strand transfer inhibitors. Clinically relevant HIV integrase resistance-associated mutations were selected in SIV in our tissue culture experiments. Not only do we report on the characterization of SIV recombinant integrase enzyme catalytic activities, we also provide the first research anywhere on the effect of mutations within recombinant integrase SIV enzymes on drug resistance.T he limitations of current highly active antiretroviral therapy (HAART) include the incidence of drug resistance observed in patients, issues of drug adherence, and numbers of newly acquired infections. The emergence of drug-resistant viruses can lead to increases in viral load and treatment failure, and such viruses can be transmitted to both healthy individuals as well as to untreated HIV-positive individuals, therefore threatening the long-term effica...

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“…Thus cross-resistance exists between RAL and EVG in regard to mutations at positions 155 and 148. In contrast, DTG has a higher barrier to HIV drug resistance than do RAL and EVG (White et al 2014;Llibre et al 2015;Seki et al 2015;Wainberg and Han 2015). The major mutations that confer resistance to RAL, EVG, and DTG are summarized in Table 1.…”

Section: The Unique Hiv Drug Resistance Profile Of Dtgmentioning

confidence: 99%

Might dolutegravir be part of a functional cure for HIV?

2016

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Antiretroviral therapy (ART) has greatly decreased HIV-related morbidity and mortality. However, HIV can establish viral reservoirs that evade both the immune system and ART. Dolutegravir (DTG) is a secondgeneration integrase strand transfer inhibitor (INSTI) related to the first-generation INSTIs raltegravir (RAL) and elvitegravir (EVG). DTG shows a higher genetic barrier to the development of HIV-1 resistance than RAL and EVG. More interestingly, clinical resistance mutations to DTG in treatment-naïve patients have not been observed to date. This review summarizes recent studies on strategies toward a cure for HIV, explores resistance profiles of DTG, and discusses how DTG might help in finding a functional cure for HIV.Key words: dolutegravir, drug resistance, HIV, cure. Résumé :Les traitements antirétroviraux (TAR) ont considérablement réduit la morbidité et la mortalité associées au VIH. Or, le VIH peut établir des réservoirs viraux pour se soustraire au système immunitaire et aux TAR. Le dolutégravir (DTG) est un inhibiteur du transfert de brin par l'intégrase (integrase strand transfer inhibitor, INSTI) de deuxième génération qui s'apparente au raltégravir (RAL) et à l'elvitégravir (EVG), des INSTI de première génération. Comparativement au RAL et à l'EVG, le DTG présente une plus grande barrière génétique au dével-oppement de la résistance chez le VIH. On remarque avec intérêt que des mutations menant à la résistance clinique au DTG chez des patients jamais traités auparavant n'ont pas encore été observées. La présente revue fait la synthèse des études récentes abordant des stratégies visant à éliminer le VIH, explore les profils de résistance au DTG et discute de la possibilité de mettre au point un remède contre le VIH à l'aide du DTG. [Traduit par la Rédaction]

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Effects of Raltegravir or Elvitegravir Resistance Signature Mutations on the Barrier to Dolutegravir Resistance In Vitro

Cited by 32 publications

References 34 publications

Dolutegravir-Selected HIV-1 Containing the N155H and R263K Resistance Substitutions Does Not Acquire Additional Compensatory Mutations under Drug Pressure That Lead to Higher-Level Resistance and Increased Replicative Capacity

Dolutegravir-Selected HIV-1 Containing the N155H and R263K Resistance Substitutions Does Not Acquire Additional Compensatory Mutations under Drug Pressure That Lead to Higher-Level Resistance and Increased Replicative Capacity

Characterization of the Drug Resistance Profiles of Integrase Strand Transfer Inhibitors in Simian Immunodeficiency Virus SIVmac239

Might dolutegravir be part of a functional cure for HIV?

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