Consideration of Metabolite Efflux in Radiolabelled Choline Kinetics

Hypoxia is a complex microenvironmental condition known to regulate choline kinase α (CHKA) activity and choline transport through transcription factor hypoxia-inducible factor-1α (HIF-1α) and, therefore, may confound the uptake of choline radiotracer [F]fluoromethyl-[1,2-H]-choline ([F]-D4-FCH). The aim of this study was to investigate how hypoxia affects the choline radiotracer dynamics. Three underlying mechanisms by which hypoxia could potentially alter the uptake of the choline radiotracer, [F]-D4-FCH, were investigated: F-D4-FCH import, CHKA phosphorylation activity, and the efflux of [F]-D4-FCH and its phosphorylated product [F]-D4-FCHP. The effects of hypoxia on [F]-D4-FCH uptake were studied in CHKA-overexpressing cell lines of prostate cancer, PC-3, and breast cancer MDA-MB-231 cells. The mechanisms of radiotracer efflux were assessed by the cell uptake and immunofluorescence in vitro and examined in vivo ( = 24). The mathematical modelling methodology was further developed to verify the efflux hypothesis using [F]-D4-FCH dynamic PET scans from non-small cell lung cancer (NSCLC) patients ( = 17). We report a novel finding involving the export of phosphorylated [F]-D4-FCH and [F]-D4-FCHP via HIF-1α-responsive efflux transporters, including ABCB4, when the HIF-1α level is augmented. This is supported by a graphical analysis of human data with a compartmental model (M2T6k + k) that accounts for the efflux. Hypoxia/HIF-1α increases the efflux of phosphorylated radiolabelled choline species, thus supporting the consideration of efflux in the modelling of radiotracer dynamics.