Impact of Primary Elvitegravir Resistance-Associated Mutations in HIV-1 Integrase on Drug Susceptibility and Viral Replication Fitness

Abram, Michael E.; Hluhanich, Rebecca; Goodman, Derrick; Andreatta, Kristen; Margot, Nicolas; Ye, Linda; Niedziela-Majka, Anita; Barnes, Tiffany; Novikov, Nikolai; Chen, Xiaowu; Svarovskaia, Evguenia S.; McColl, Damian J.; White, Kirsten; Miller, Michael D.

doi:10.1128/aac.02568-12

Cited by 102 publications

(88 citation statements)

References 47 publications

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“…In contrast, the combination of H51Y and R263K in SIV conferred low-level resistance against both DTG and EVG in our study (1.4-fold and 1.7-fold, respectively) ( Table 4). The E92Q variant enzyme displayed low-level resistance against EVG, consistent with results obtained for HIV-1 (31,57), but the combination of E92Q and T97A did not alter this resistance profile, nor did E92Q T97A confer resistance against either RAL or EVG (Table 4). The HIV RAL resistance-associated substitution, Y143R, also conferred resistance to SIV against this compound but not against DTG or EVG ( Table 4).…”

Section: Discussionsupporting

confidence: 76%

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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Section: Discussionsupporting

confidence: 76%

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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“…One study showed that the addition of E92Q or G163R to N155H was able to increase viral fitness in both the absence and presence of RAL, whereas the accessory mutation L74M increased replication capacity only in the presence of the drug (14). A different study showed that the E92Q substitution in combination with N155H was deleterious to HIV-1 replication in the absence of INSTI pressure (15). The E157Q substitution has been shown to have variable effects on INSTI activity (16,17), while a T97A substitution has been reported to cause a 70% decrease in viral replication relative to that of the WT (15).…”

Section: Discussionmentioning

confidence: 99%

“…A different study showed that the E92Q substitution in combination with N155H was deleterious to HIV-1 replication in the absence of INSTI pressure (15). The E157Q substitution has been shown to have variable effects on INSTI activity (16,17), while a T97A substitution has been reported to cause a 70% decrease in viral replication relative to that of the WT (15).…”

Section: Discussionmentioning

confidence: 99%

“…It is generally agreed that the five accessory mutations discussed here contribute to higher levels of resistance to the older INSTIs RAL and EVG than do the primary mutations for these two drugs on their own (14,15,19,20). In this study, we have expanded on these results by showing that each triply mutated virus is just as or more resistant to each INSTI than the N155H/ R263K double mutant, with the exception of L74M/N155H/ R263K for DTG and N155H/G163R/R263K for RAL.…”

Section: Discussionmentioning

confidence: 99%

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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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“…It is noteworthy that the replicative as well as transmission properties of mutant and wild type variants are highly different. For example, certain data suggest that some drug resistant variants possess reduced replicative capacity in comparison with wild-type virus and therefore, drug resistant infection might be of some virological benefit to the hosts' immune system [39][40][41] .…”

Section: History and Mechanisms Of Anti-retroviral Drug Resistance (Amentioning

confidence: 99%

The emergence of drug resistant HIV variants and novel anti–retroviral therapy

Paydary

Khaghani

Emamzadehfard

et al. 2013

Asian Pacific Journal of Tropical Biomedicine

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Impact of Primary Elvitegravir Resistance-Associated Mutations in HIV-1 Integrase on Drug Susceptibility and Viral Replication Fitness

Cited by 102 publications

References 47 publications

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

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

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

The emergence of drug resistant HIV variants and novel anti–retroviral therapy

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