In vitro and patient studies with platelets to explore off‐target cardiovascular effects of integrase inhibitors

Figures

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  FIGURE 1. Platelet aggregation upon in vitro treatment with bictegravir (BIC) and dolutegravir (DTG). Panel (a) shows percentage platelet aggregation from N = 5 upon exposure to peak plasma concentration (Cmax) of BIC, DTG, or dimethylsulfoxide (DMSO) for 25 min followed by adenosine 5′‐diphosphate (ADP) activation using a range of agonist concentrations from 1 to 30 μM ADP. Similarly, (b) shows thrombin receptor activator for peptide 6 (TRAP‐6)‐evoked platelet aggregation using 1–30 μM TRAP‐6, and (c) shows collagen‐induced aggregation ranging from 1 to 30 μg/mL of collagen. Results were analysed using one‐way analysis of variance, and groups were compared using Tukey's multiple comparison: * p < 0.05, ** p < 0.01, *** p < 0.001.

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  FIGURE 2. Eptifibatide‐inhibited platelet aggregation after treatment with bictegravir (BIC) or dolutegravir (DTG). Platelets from volunteers without HIV (N = 4) were treated with clinically relevant concentrations of BIC, DTG, or vehicle control (dimethylsulfoxide [DMSO]) as well as with 0.6 μg/mL of eptifibatide, a specific αΙΙbβ3 inhibitor, to block integrin‐mediated platelet aggregation. The graphs show (a) 3 μM adenosine 5′‐diphosphate (ADP), (b) 10 μM thrombin receptor activator for peptide 6 (TRAP‐6), and (c) 3 μg/mL collagen‐evoked platelet aggregation, 25 min after treatment with the drugs ± eptifibatide. Results were analysed using two‐way analysis of variance, and groups were compared using Tukey's multiple comparison: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

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  FIGURE 3. Aggregation profiles of bictegravir (BIC)/emtricitabine (FTC)/tenofovir alafenamide fumarate (TAF)‐ or dolutegravir (DTG)/lamivudine (3TC)‐treated platelets. Top row (a) shows aggregation profiles of adenosine 5′‐diphosphate (ADP)‐evoked platelet activation at increasing agonist concentrations from left to right, after in vitro treatment of platelets from healthy volunteers (N = 5) with clinically relevant concentrations of BIC/FTC/TAF, DTG/3TC, or vehicle (dimethylsulfoxide). Row (b) indicates thrombin receptor activator for peptide 6 (TRAP‐6)‐induced and row (c) the collagen‐induced platelet aggregation at increasing concentrations from left to right. Results were statistically analysed using one‐way analysis of variance, and groups were compared using Tukey's multiple comparison: *p < 0.05, **p < 0.01, ***p < 0.001.

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  FIGURE 4. Fold change in CD62P and CD63 levels after in vitro treatment of platelets with bictegravir (BIC) or dolutegravir (DTG). The graph shows (a) the fold change in CD62P (P‐selectin) in platelet‐rich plasma (N = 6) treated with BIC, DTG, or vehicle control (dimethylsulfoxide [DMSO]) after 10 μM adenosine 5′‐diphosphate (ADP), 10 μM thrombin receptor activator for peptide 6 (TRAP‐6), or 10 μg/mL collagen activation, in flow cytometry. Graph (b) shows the fold change in CD63. Results from N = 6 were analysed with two‐way analysis of variance, and groups were compared using Tukey's multiple comparison test: *p < 0.05.

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  FIGURE 5. Aggregation profiles of platelets from people living with HIV on bictegravir (BIC)/emtricitabine (FTC)/tenofovir alafenamide fumarate (TAF) or dolutegravir (DTG)/lamivudine (3TC). The figure shows (a) the adenosine 5′‐diphosphate (ADP)‐induced platelet aggregation in platelet‐rich plasma (PRP) from people with HIV, N = 8 on BIC/FTC/TAF and N = 8 ON DTG/3TC. Row (b) demonstrates thrombin receptor activator for peptide 6 (TRAP‐6)‐induced aggregation in PRP from people with HIV in the two treatment groups, and row (c) the collagen‐induced aggregation. Results are analysed using two‐way analysis of variance, and groups were compared with Tukey's multiple comparison. No statistically significant differences were observed.