Thibault Mesplède1, Nathan Osman2, Melissa Wares2, Peter K Quashie3, Said Hassounah3, Kaitlin Anstett2, Yingshan Han1, Diane N Singhroy2, Mark A Wainberg4. 1. McGill University AIDS Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, Québec, Canada. 2. McGill University AIDS Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, Québec, Canada Department of Microbiology and Immunology, Faculty of Medicine, McGill University, Montréal, Québec, Canada. 3. McGill University AIDS Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, Québec, Canada Division of Experimental Medicine, Faculty of Medicine, McGill University, Montréal, Québec, Canada. 4. McGill University AIDS Centre, Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, Québec, Canada Department of Microbiology and Immunology, Faculty of Medicine, McGill University, Montréal, Québec, Canada Division of Experimental Medicine, Faculty of Medicine, McGill University, Montréal, Québec, Canada mark.wainberg@mcgill.ca.
Abstract
BACKGROUND: The results of several clinical trials suggest that the integrase inhibitor dolutegravir may be less prone than other drugs to the emergence of HIV drug resistance mutations in treatment-naive patients. We have shown that the R263K mutation commonly emerged during tissue culture selection studies with dolutegravir and conferred low levels of resistance to this drug while simultaneously diminishing both HIV replication capacity and integrase enzymatic activity. E138K has been identified as a secondary mutation for dolutegravir in selection studies and has also been observed as a secondary mutation in the clinic for the integrase inhibitors raltegravir and elvitegravir. METHODS: We used biochemical cell-free strand-transfer assays and tissue culture assays to characterize the effects of the E138K/R263K combination of mutations on resistance to dolutegravir, integrase enzyme activity and HIV-1 replication capacity. RESULTS: We show here that the addition of the E138K substitution to R263K increased the resistance of HIV-1 to dolutegravir but failed to restore viral replication capacity, integrase strand-transfer activity and integration within cellular DNA. We also show that the addition of E138K to R263K did not increase the resistance to raltegravir or elvitegravir. The addition of the E138K substitution to R263K was also less detrimental to integrase strand-transfer activity and integration than a different secondary mutation at position H51Y that had also been selected in culture. CONCLUSIONS: The E138K substitution failed to restore the defect in viral replication capacity that is associated with R263K, confirming previous selection studies that failed to identify compensatory mutation(s) for the latter primary mutation. This study suggests that the R263K resistance pathway may represent an evolutionary dead end for HIV in treatment-naive individuals who are treated with dolutegravir and will need to be confirmed by the long-term use of dolutegravir in the clinic.
BACKGROUND: The results of several clinical trials suggest that the integrase inhibitor dolutegravir may be less prone than other drugs to the emergence of HIV drug resistance mutations in treatment-naive patients. We have shown that the R263K mutation commonly emerged during tissue culture selection studies with dolutegravir and conferred low levels of resistance to this drug while simultaneously diminishing both HIV replication capacity and integrase enzymatic activity. E138K has been identified as a secondary mutation for dolutegravir in selection studies and has also been observed as a secondary mutation in the clinic for the integrase inhibitors raltegravir and elvitegravir. METHODS: We used biochemical cell-free strand-transfer assays and tissue culture assays to characterize the effects of the E138K/R263K combination of mutations on resistance to dolutegravir, integrase enzyme activity and HIV-1 replication capacity. RESULTS: We show here that the addition of the E138K substitution to R263K increased the resistance of HIV-1 to dolutegravir but failed to restore viral replication capacity, integrase strand-transfer activity and integration within cellular DNA. We also show that the addition of E138K to R263K did not increase the resistance to raltegravir or elvitegravir. The addition of the E138K substitution to R263K was also less detrimental to integrase strand-transfer activity and integration than a different secondary mutation at position H51Y that had also been selected in culture. CONCLUSIONS: The E138K substitution failed to restore the defect in viral replication capacity that is associated with R263K, confirming previous selection studies that failed to identify compensatory mutation(s) for the latter primary mutation. This study suggests that the R263K resistance pathway may represent an evolutionary dead end for HIV in treatment-naive individuals who are treated with dolutegravir and will need to be confirmed by the long-term use of dolutegravir in the clinic.
Authors: Jan van Lunzen; Franco Maggiolo; José R Arribas; Aza Rakhmanova; Patrick Yeni; Benjamin Young; Jürgen K Rockstroh; Steve Almond; Ivy Song; Cindy Brothers; Sherene Min Journal: Lancet Infect Dis Date: 2011-10-20 Impact factor: 25.071
Authors: Jürgen K Rockstroh; Edwin DeJesus; Jeffrey L Lennox; Yazdan Yazdanpanah; Michael S Saag; Hong Wan; Anthony J Rodgers; Monica L Walker; Michael Miller; Mark J DiNubile; Bach-Yen Nguyen; Hedy Teppler; Randi Leavitt; Peter Sklar Journal: J Acquir Immune Defic Syndr Date: 2013-05-01 Impact factor: 3.731
Authors: Edwin DeJesus; Jürgen K Rockstroh; Keith Henry; Jean-Michel Molina; Joseph Gathe; Srinivasan Ramanathan; Xuelian Wei; Kitty Yale; Javier Szwarcberg; Kirsten White; Andrew K Cheng; Brian P Kearney Journal: Lancet Date: 2012-06-30 Impact factor: 79.321
Authors: Martin Markowitz; Bach-Yen Nguyen; Eduardo Gotuzzo; Fernando Mendo; Winai Ratanasuwan; Colin Kovacs; Guillermo Prada; Javier O Morales-Ramirez; Clyde S Crumpacker; Robin D Isaacs; Lucinda R Gilde; Hong Wan; Michael D Miller; Larissa A Wenning; Hedy Teppler Journal: J Acquir Immune Defic Syndr Date: 2007-10-01 Impact factor: 3.731
Authors: Martin Markowitz; Bach-Yen Nguyen; Eduardo Gotuzzo; Fernando Mendo; Winai Ratanasuwan; Colin Kovacs; Guillermo Prada; Javier O Morales-Ramirez; Clyde S Crumpacker; Robin D Isaacs; Havilland Campbell; Kim M Strohmaier; Hong Wan; Robert M Danovich; Hedy Teppler Journal: J Acquir Immune Defic Syndr Date: 2009-11-01 Impact factor: 3.731
Authors: Melissa Wares; Said Hassounah; Thibault Mesplède; Paul A Sandstrom; Mark A Wainberg Journal: Antimicrob Agents Chemother Date: 2015-01-12 Impact factor: 5.191
Authors: Emmanuel Ndashimye; Mariano Avino; Fred Kyeyune; Immaculate Nankya; Richard M Gibson; Eva Nabulime; Art F Y Poon; Cissy Kityo; Peter Mugyenyi; Miguel E Quiñones-Mateu; Eric J Arts Journal: AIDS Res Hum Retroviruses Date: 2018-02-26 Impact factor: 2.205