Comparative Evaluation of [¹⁸F]5-Fluoroaminosuberic Acid and (4<i>S</i>)-4-3-[¹⁸F]fluoropropyl)-l-Glutamate as System xC--Targeting Radiopharmaceuticals.

Abstract

System [Formula: see text] is an appealing biomarker for targeting oxidative stress with oncologic PET imaging and can serve as an alternative PET biomarker to other metabolic indicators. In this paper, we report a direct comparison of 2 ¹⁸F-labeled amino acid radiopharmaceuticals targeting system [Formula: see text], [¹⁸F]5-fluoroaminosuberic acid ([¹⁸F]FASu) and (4<i>S</i>)-4-(3-[¹⁸F]fluoropropyl)-l-glutamate ([¹⁸F]FSPG), in terms of their uptake specificity and ability to image glioma and lung cancer xenografts in vivo. <b>Methods:</b> Both tracers were synthesized according to previously published procedures. In vitro uptake specificity assays were conducted using prostate (PC-3), glioblastoma (U-87), colorectal (HT-29), ovarian (SKOV3), breast (MDA-MB-231), and lung cancer (A549) cell lines. PET/CT imaging and biodistribution studies were conducted in immunocompromised mice bearing U-87 or A549 xenografts. <b>Results:</b> In vitro cell uptake assays showed that the tracers accumulated in cancer cells in a time-dependent manner and that the uptake of [¹⁸F]FASu was blocked by the system [Formula: see text] inhibitor sulfasalazine and rose bengal, but not by system L inhibitor 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, system [Formula: see text] inhibitor L-<i>trans</i>-pyrrolidine-2,4-dicarboxylic acid, or l-serine, which is a substrate for transporter systems A, ACS, B⁰, and B^0,+ Conversely, [¹⁸F]FSPG uptake decreased significantly in the presence of an excess of L-<i>trans</i>-pyrrolidine-2,4-dicarboxylic acid in 2 of 3 tested cell lines, indicating some reliance on system [Formula: see text] in these cells. In an in vivo setting, [¹⁸F]FASu and [¹⁸F]FSPG generated good-contrast PET images in U-87 and A549 tumor-bearing mice. Tracer accumulation in A549 tumors was 5.0 ± 0.8 percentage injected dose (%ID)/g ([¹⁸F]FASu, <i>n</i> ≥ 5) and 6.3 ± 1.3 %ID/g ([¹⁸F]FSPG, <i>n</i> ≥ 6, <i>P</i> = 0.7786), whereas U-87 xenografts demonstrated uptake of 6.1 ± 2.4 %ID/g ([¹⁸F]FASu, <i>n</i> ≥ 4) and 11.2 ± 4.1 %ID/g ([¹⁸F]FSPG, <i>n</i> ≥ 4, <i>P</i> = 0.0321) at 1 h after injection. <b>Conclusion:</b> [¹⁸F]FSPG had greater in vitro uptake than [¹⁸F]FASu in all cell lines tested; however, our results indicate that residual uptake differences exist between [¹⁸F]FSPG and [¹⁸F]FASu, suggesting alternative transporter activity in the cell lines tested. In vivo studies demonstrated the ability of both [¹⁸F]FASu and [¹⁸F]FSPG to image glioblastoma (U-87) and non-small cell lung cancer (A549) xenografts.