Primary mutations selected in vitro with raltegravir confer large fold changes in susceptibility to first-generation integrase inhibitors, but minor fold changes to inhibitors with second-generation resistance profiles

Goethals, Olivia; Vos, Ann; Ginderen, Marcia Van; Geluykens, Peggy; Smits, Veerle; Schols, Dominique; Hertogs, Kurt; Clayton, R. M.

doi:10.1016/j.virol.2010.03.034

Cited by 58 publications

(46 citation statements)

References 41 publications

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“…The level of resistance provided by these pathways during the infection of T cells has been investigated previously (19,46). However, fundamental differences in cellular composition, metabolism, and viral replication kinetics can lead to differences in the efficacy of antiviral drugs in T cells and macrophages (3).…”

Section: Stimulated Cd4mentioning

confidence: 99%

Single Mutations in HIV Integrase Confer High-Level Resistance to Raltegravir in Primary Human Macrophages

Marsden

Avancena

Kitchen

et al. 2011

Antimicrob Agents Chemother

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CD4؉ T cells and macrophages are the primary target cells for HIV in vivo, and antiretroviral drugs can vary in their ability to inhibit the infection of these different cell types. Resistance pathways to the HIV integrase inhibitor raltegravir have previously been investigated in T cells. Primary raltegravir resistance mutations, most often at integrase amino acid position 148 or 155, afford some resistance to the drug. The acquisition of pathway-specific secondary mutations then provides higher-level resistance to viruses infecting T cells. We show here that during macrophage infection, the presence of a single primary raltegravir resistance mutation (Q148H, Q148R, N155H, or N155S) is sufficient to provide resistance to raltegravir comparable to that seen in viruses expressing both primary and secondary mutations in costimulated CD4؉ T cells. These data implicate macrophages as a potential in vivo reservoir that may facilitate the development of resistance to raltegravir. Notably, the newer integrase inhibitor MK-2048 effectively suppressed the infection of all raltegravir-resistant viruses in both T cells and macrophages, indicating that more recently developed integrase inhibitors are capable of inhibiting infection in both major HIV cellular reservoirs, even in patients harboring raltegravir-resistant viruses.

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Section: Stimulated Cd4mentioning

confidence: 99%

Single Mutations in HIV Integrase Confer High-Level Resistance to Raltegravir in Primary Human Macrophages

Marsden

Avancena

Kitchen

et al. 2011

Antimicrob Agents Chemother

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“…Numerous secondary mutations confer low levels of resistance against both drugs (reviewed in reference 47). Secondgeneration INSTIs have been developed, which possess a more robust resistance profile than RAL and EVG (3,21,27,33,67). These include MK-2048 (3, 4, 20, 67) and dolutegravir (DTG) (3,20,33,45,46,61,67).…”

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confidence: 99%

Characterization of the R263K Mutation in HIV-1 Integrase That Confers Low-Level Resistance to the Second-Generation Integrase Strand Transfer Inhibitor Dolutegravir

Quashie

Mésplède

Han

et al. 2012

J Virol

214

306

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Integrase (IN) strand transfer inhibitors (INSTIs) have been developed to inhibit the ability of HIV-1 integrase to irreversibly link the reverse-transcribed viral DNA to the host genome. INSTIs have proven their high efficiency in inhibiting viral replication in vitro and in patients. However, first-generation INSTIs have only a modest genetic barrier to resistance, allowing the virus to escape these powerful drugs through several resistance pathways. Second-generation INSTIs, such as dolutegravir (DTG, S/GSK1349572), have been reported to have a higher resistance barrier, and no novel drug resistance mutation has yet been described for this drug. Therefore, we performed in vitro selection experiments with DTG using viruses of subtypes B, C, and A/G and showed that the most common mutation to emerge was R263K. Further analysis by site-directed mutagenesis showed that R263K does confer low-level resistance to DTG and decreased integration in cell culture without altering reverse transcription. Biochemical cell-free assays performed with purified IN enzyme containing R263K confirmed the absence of major resistance against DTG and showed a slight decrease in 3= processing and strand transfer activities compared to the wild type. Structural modeling suggested and in vitro IN-DNA binding assays show that the R263K mutation affects IN-DNA interactions.T he high mutation rate of HIV-1 reverse transcriptase (RT) allows the virus to escape pressure through adaptive mutations that include drug resistance mutations that limit the effectiveness of antiretroviral drugs (5,31,66,69,70). The use of multiple drugs in combination can hamper this process by restraining viral replication, limiting the emergence of resistant strains. The addition of integrase inhibitors to the arsenal of drugs against HIV-1 is important since these inhibitors are active against viruses resistant to other drug classes (16,49,63).The HIV-1 integrase enzyme catalyzes two reactions. The first is 3= processing, which consists of cleavage of a dinucleotide at both 3= ends of the reverse-transcribed linear viral DNA and results in the exposure of reactive hydroxyl groups. The second step termed "strand transfer" is carried out through a nucleophilic attack by exposed 3= hydroxyl groups on host genomic DNA (26, 47). Even though 3= processing may be a suitable therapeutic target, the integrase inhibitors developed so far are integrase strand transfer inhibitors (INSTIs) that preferentially inhibit strand transfer while only modestly affecting 3= processing (18,24,26). Raltegravir (RAL) was the first INSTI to be approved for therapy in 2007 (64) and is safe and efficient in both treatment-naïve and treatment-experienced subjects (11,17,23,35,49,62,63). Elvitegravir (EVG) is another INSTI currently in advanced clinical trials (10,12,77).Although first-generation INSTIs strongly inhibit HIV-1 replication, they possess only a modest genetic barrier to resistance. Three main resistance pathways have been identified for RAL, involving initial mutations of the N1...

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“…The N155H mutation was usually accompanied by the secondary mutation L74M, E92Q, T97A, G136R, or V151I (23)(24)(25). Although secondary mutations do not individually confer INSTI resistance, their combinations with N155H increase resistance and restore viral replication capacity (26,27). To assess whether mutations at L74 alone or with V75I act as secondary mutations, we evaluated resistance level (Fig.…”

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confidence: 99%

Impact of HIV-1 Integrase L74F and V75I Mutations in a Clinical Isolate on Resistance to Second-Generation Integrase Strand Transfer Inhibitors

Hachiya

Kirby

Ido

et al. 2017

Antimicrob Agents Chemother

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A novel HIV-1 integrase mutation pattern, L74F V75I, which conferred resistance to first-generation integrase strand transfer inhibitors (INSTIs), was identified in a clinical case with virological failure under a raltegravir-based regimen. Addition of L74F V75I to N155H or G140S Q148H increased resistance levels to the second-generation INSTIs dolutegravir (Ͼ385-and 100-fold, respectively) and cabotegravir (153-and 197-fold, respectively). These findings are important for the development of an accurate system for interpretation of INSTI resistance and the rational design of next-generation INSTIs.KEYWORDS dolutegravir, drug resistance mechanisms, human immunodeficiency virus, integrase, integrase strand transfer inhibitor I ntegrase strand transfer inhibitors (INSTIs) constitute the latest class of available antiretroviral agents exhibiting potent antiretroviral effects in vitro and vivo (1-8). The first-generation INSTIs raltegravir (RAL) and elvitegravir (EVG) display broad crossresistance, whereas second-generation INSTIs of carbamoyl pyridine analogues, dolutegravir (DTG) and cabotegravir (CAB), demonstrate superior activity against firstgeneration INSTI-resistant HIV-1 variants (4, 9-13). Although a DTG resistance mutation, R263K, has been reported, and its resistance mechanism has been well studied (10,12,14,15), other potential DTG resistance mutations and their mechanisms are not fully understood.In a clinical case (Fig. 1A), the plasma HIV-1 RNA level rebounded to 2,900 copies/ml 1 year after starting antiretroviral therapy (ART) that included tenofovir disoproxil fumarate (TDF), emtricitabine (FTC), and RAL. However, no known INSTI resistance mutations were identified based on major drug resistance mutation lists (IAS-USA drug resistance mutations list [16] and the HIV Drug Resistance Database at Stanford University [HIVDB]) at time points 2 and 3 (Fig. 1B). Clinical samples were obtained from the fresh plasma of a patient attending the outpatient clinic of the National Hospital Organization Nagoya Medical Center. The Institutional Review Board approved this study , and written informed consent was obtained from this patient. To identify novel mutations associated with RAL resistance in the clinical isolates, we constructed infectious HIV-1 clones with cDNA fragments of the integrase (IN)-coding region derived from the clinical isolates and performed phenotypic resistance assays using TZM-bl cells as previously described (8, 17). Briefly, viral RNA was extracted from

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Primary mutations selected in vitro with raltegravir confer large fold changes in susceptibility to first-generation integrase inhibitors, but minor fold changes to inhibitors with second-generation resistance profiles

Cited by 58 publications

References 41 publications

Single Mutations in HIV Integrase Confer High-Level Resistance to Raltegravir in Primary Human Macrophages

Single Mutations in HIV Integrase Confer High-Level Resistance to Raltegravir in Primary Human Macrophages

Characterization of the R263K Mutation in HIV-1 Integrase That Confers Low-Level Resistance to the Second-Generation Integrase Strand Transfer Inhibitor Dolutegravir

Impact of HIV-1 Integrase L74F and V75I Mutations in a Clinical Isolate on Resistance to Second-Generation Integrase Strand Transfer Inhibitors

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