Fatty Acid-Binding Protein 5 Mediates the Uptake of Fatty Acids, but not Drugs, Into Human Brain Endothelial Cells

The purpose of this study was to examine the involvement of fatty acid-binding protein 5 (FABP5), a lipid-binding protein expressed at the blood-brain barrier (BBB), in fatty acid and drug uptake into human brain endothelial cells. Following transfection with siRNA against hFABP5, human brain endothelial cell (hCMEC/D3) uptake of lipophilic ligands with varying affinity to FABP5 was assessed with intracellular concentrations quantified by liquid scintillation counting, HPLC, or LCMS/MS. The in situ BBB transport of [H-3]-diazepam was also assessed in wild type and FABP5-deficient mice. hFABP5 siRNA reduced FABP5 expression in hCMEC/D3 cells by 39.9 +/- 3.8% (mRNA) and 38.8 +/- 6.6% (protein; mean +/- SEM), leading to a reduction in uptake of [C-14]-lauric acid, [H-3]-oleic acid, and [C-14]-stearic acid by 37.5 +/- 8.8%, 41.7 +/- 11.6%, and 50.7 +/- 13.6%, respectively, over 1 min. No significant changes in [C-14]-diazepam, pioglitazone, and troglitazone uptake were detected following FABP5 knockdown in hCMEC/D3 cells. Similarly, no difference in BBB transport of [3H]-diazepam was observed between wild type and FABP5-deficient mice. Therefore, although FABP5 facilitates brain endothelial cell uptake of fatty acids, it has limited effects on brain endothelial cell uptake and BBB transport of drugs with lower affinity for FABP5. Crown Copyright (C) 2018 Published by Elsevier Inc. on behalf of the American Pharmacists Association. All rights reserved.