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| Hauptseite > Publikationsdatenbank > Evaluation of a multifunctional blood-brain barrier co-culture model prepared from rat primary brain endothelial cells and astrocytes: first results |
| Hauptseite > Publikationsdatenbank > Evaluation of a multifunctional blood-brain barrier co-culture model prepared from rat primary brain endothelial cells and astrocytes: first results |
| Conference Presentation (After Call) | FZJ-2024-04010 |
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2022
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Please use a persistent id in citations: doi:10.1016/S0969-8051(22)00233-5
Abstract: Objectives: The potential of a rodent endothelial cell/astrocyteco-culture BBB model to predict in vivo brain exposure ofCNS radiotracers was evaluated using three 18F-labeled xanthine-derived positron emission tomography (PET) tracers foradenosine A1 receptor (A1AR) imaging, namely 8-cyclopentyl-3-(3-[18F]Fluoropropyl)-1-propylxanthine ([18F]CPFPX [1,2]), 8-cyclobutyl-3-(3-[18F]Fluoropropyl)-1-propylxanthine ([18F]CBX [3]), and3-(3-[18F]Fluoropropyl)-8-(1-methylcyclobutyl)-1-propylxanthine([18F]MCBX [3]).Methods: Primary rat brain astrocytes (4x104 cells) and primaryrat brain microvascular endothelial cells (BECs, 2x104 cells) wereplated on opposite sides of a polycarbonate Transwell membrane(pore size 3.0 m) and cultured at 37°C in a 5% CO2 atmosphere for4 days [4]. Integrity of the endothelial cell layer was assessed bytransendothelial electrical resistance (TEER) measurements. Forpermeability testing, the radiolabeled compounds (185 kBq/ml)were added to the donor chamber (apical compartment), and theirappearance in the receiver chamber (basolateral compartment) wasmonitored with a gamma counter to calculate apparent permeabilities(Papp). Papp values of the xanthines were compared to Pin vivo valuesderived from PET kinetic modeling data [5]. Additional experimentswere conducted to investigate the influence of test compound concentrationand addition of albumin on Papp values.Results: Figure 1 displays time dependent permeation of [18F]CPFPX, [18F]CBX, and [18F]MCBX across the model BBB. Calculated Pappvalues and the corresponding Pin vivo values obtained from PET measurementsare listed in Table 1. Papp values proved to be highly predictivefor in vivo brain penetration. Permeability rankings in vivo andin vitro were comparable ([18F]MCBX > [18F]CBX≈[18F]CPFPX).Papp values of [18F]CPFPX did not show concentration dependence,indicating that passage of the compound through the BBB proceedssolely via transmembrane diffusion without involvement of saturabletransport mechanism. However, addition of bovine serumalbumin (30 mg/ml, corresponding to a free ligand concentration of7%) significantly lowered Papp of [18F]CPFPX by about 30%, which isconsistent with the free drug hypothesis.Conclusions: This first study demonstrates a strong agreementbetween in vitro cell-based permeability data and in vivo brain penetrationmeasured by PET. If these results can be confirmed withother classes of molecules that exhibit different transport characteristicsat the BBB (e.g., P-glycoprotein substrates), the BBB modeldescribed here should prove valuable for the development of novelCNS radiotracers.Acknowledgments: We thank Prof. Dr. Dieter Willbold andDominik Honold from the Institute of Structural Biochemistry (IBI-7)at FZJ for their valuable support.References:[1] Holschbach et al., J Med Chem. 2002, 45(23), 5150-5156.[2] Bauer et al., J Nucl Med. 2003, 44(10), 1682-1689.[3] Schneider et al., Pharmaceuticals. 2019, 12(2), 57.[4] Niego et al., J Vis Exp. 2013, 81, e50934.[5] Schneider et al., Nucl Med Biol. 2020, 82-83, 1-8.
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