N. Ghazal,1 Y. Yeung,2 Z. Ngan,2 H. Wang,2 M. El-Sakkary,2 F. Belanger,1 N. Sheehan,3 B. Lebouche,3 L. Labbe,2 J. Turgeon,1 V. Michaud1; 1CHUM Research Centre, Montreal, QC, Canada, 2Faculty of Pharmacy, University of Montreal, Montreal, QC, Canada, 3McGill University Health Center, Montréal, QC, Canada
BACKGROUND: The pharmacological target of most antiretroviral (ARV) agents is within cells infected with HIV. Clinical outcomes depend on intracellular (IC) drug concentrations which, for ARV, can be influenced by drug influx and efflux transporters. Diabetes is associated with inflammatory components that can affect protein expression including that of drug-transporters. Our preliminary objective was to investigate the impact of diabetes on mRNA drug-transporters in peripheral blood mononuclear cells (PBMCs) from HIV-infected patients treated with atazanavir/ritonavir, darunavir/ritonavir or tenofovir-based regimens.
METHODS: Blood sample specimens were obtained from HIV-infected patients with diabetes (n=9) and without diabetes (n=12) under an approved research protocol. PBMC were isolated by density gradient centrifugation using CPT tubes. Total mRNA was extracted from PBMC pellet and q-PCR assays were performed using TaqMan probe (Applied Biosystems) for ABCB1, ABCC1-2-4-10, OCT1-2, ABCG2, the co-receptors CXCR4 and CCR5 and GAPDH. Relative mRNA expression was determined using the ΔΔCT methods.
RESULTS: No statistical difference in mRNA levels was observed among HIV-infected patients with diabetes compared to non-diabetics for all tested drug-transporters and co-receptors. ABCC10, OCT2 and CCR5 mRNAs tended to be higher in the diabetic group while ABCC4 tended to be higher in the non-diabetic group. Expression was highly variable, especially among diabetic HIV-infected patients (CV114-185%). Drug-transporters with the lowest and the highest average expression in PBMC were ABCG2 and ABCC10, respectively.
CONCLUSION: Our results need to be confirmed in a larger cohort of HIV-infected patients as well as in diabetic HIV-negative patients.