Mécanismes de résistance aux inhibiteurs de CCR5
The work described in this manuscript aimed to characterize the resistance pathways to CCR5 inhibitors. We first developed a phenotypic assay to identify viral tropism and to measure the resistance to CCR5 inhibitors. This assay detects R5 tropic viruses or X4 when they represent as little as 0.4% in a mixture of viral populations, demonstrating its robustness and sensitivity. Based on its clonal approach, this assay can differentiate the truly dual-tropic viruses from mixed viral populations. We then studied the impact of mutations in the variable regions of gp120 envelope protein of HIV-1 virus on resistance to CCR5 inhibitors. To do this, resistant viruses were generated by passage of CC1/85 and BAL isolates in the presence of sub-inhibitory concentrations of maraviroc (MVC) and vicriviroc (VCV). Following some passages of the CC1/85 virus in the presence of MVC, some mutations were identified in differents regions of the gp120. We further selected three mutations in the variable domains of gp120, V169M in V2, L317W in V3 and I408T in V4 to construct viruses containing single, double and triple mutations to assess the contribution of individual or combined mutations in the resistance phenotype to MVC and VCV. We determined the sensitivity of each mutant to MVC and VCV, the tropism and the percentage of infectivity compared to wild type. Our results showed that the sequences that carry mutations in the V4 domain I408T, had the most impact on susceptibility to MVC. Finally, we aimed to evaluate the antiviral activity of a new CCR5 inhibitor, VCH-286 and its interaction with representative drugs from different classes of antiretroviral (ARVs) such as reverse transcriptase inhibitors, protease inhibitors, integrase inhibitors and fusion inhibitors used in clinic to treat HIV/AIDS and other CCR5 inhibitors such as MVC and VVC to assess whether these drugs could be used together within the same treatment regimen. To answer this question, we first evaluated the antiviral activity of the three CCR5 inhibitors: VCH-286, MVC, and VVC. We then evaluated the interactions of VCH -286 with MVC VVC. We finally evaluated the interactions of VCH -286 with other ARV drugs These studies showed that VCH-286 is a potent inhibitor of R5 viruses with antiviral activity at the nanomolar range and favorable drug interactions with the majority of ARVs such as reverse transcriptase, protease, integrase and fusion inhibitors employed clinically to treat HIV/AIDS. The combinations of CCR5 inhibitors have interactions ranging from synergy to antagonism. Our results show that the plasticity of the viral envelope of HIV-1 affects resistance to CCR5 inhibitors, its tropism and the potential combination of these drugs.
HIV resistance HIV inhibitors coreceptor CCR5