组织环境对确定肿瘤代谢易感性起着至关重要的作用。然而，体内药物测试速度较慢，等待肿瘤生长延迟可能不是最合适的代谢治疗终点。在生理环境下，一个用于快速测量能量应激的体内方法将能够迅速确定肿瘤靶向。碘化钠共转运体（NIS）是一种成像报告基因，其蛋白产物能够共转运钠和碘离子以及正电子发射断层扫描（PET）放射标记的阴离子进入细胞。在这里，我们展示了PET成像NIS介导的放射性示踪剂摄取可以在体内治疗后几分钟内迅速可视化肿瘤的能量应激。我们改造了HEK293T人胚肾细胞，A549和H358肺癌细胞以表达转基因NIS。接下来，我们给这些细胞和移植瘤暴露于已知可以诱导代谢应激的药物，以观察对NIS活性和能量状态的影响。我们使用[18F]四氟硼酸酯正电子发射断层扫描（PET）成像在体内非侵入地成像NIS活性。HEK293T细胞体外处理需要Na+/K+ ATP酶抑制剂地高辛后，NIS活性被破坏，证实摄入放射性示踪剂依赖于钠-钾浓度梯度。纵观HEK293T细胞和A549细胞，通过抑制糖酵解和氧化磷酸化的联合作用导致的ATP再合成中断以及通过抑制氧化磷酸化引起的作用，NIS介导的放射性示踪剂摄取分别在体外显著降低（分别为-58.2%和-16.6%）。在体内，通过与氧化磷酸化抑制剂IACS-010759处理亚皮下转基因A549肿瘤，PET信号在治疗开始后90分钟内显著降低（降低了56.5%），显示NIS能够在非侵入的方式下迅速并敏感地检测到能量应激，而在能够检测到更广泛的磷酸化AMP激活的蛋白激酶、磷酸化丙酮酸脱氢酶和GLUT1的变化之前。NIS可作为对导致ATP枯竭的药物的快速代谢传感器。NIS的PET成像可以促进针对能量途径的治疗药物在体内的测试，确定药物效力，并加速代谢药物的发展。© 2023. BioMed Central Ltd., part of Springer Nature.

Tissue environment is critical in determining tumour metabolic vulnerability. However, in vivo drug testing is slow and waiting for tumour growth delay may not be the most appropriate endpoint for metabolic treatments. An in vivo method for measuring energy stress would rapidly determine tumour targeting in a physiologically relevant environment. The sodium-iodide symporter (NIS) is an imaging reporter gene whose protein product co-transports sodium and iodide, and positron emission tomography (PET) radiolabelled anions into the cell. Here, we show that PET imaging of NIS-mediated radiotracer uptake can rapidly visualise tumour energy stress within minutes following in vivo treatment.We modified HEK293T human embryonic kidney cells, and A549 and H358 lung cancer cells to express transgenic NIS. Next, we subjected these cells and implanted tumours to drugs known to induce metabolic stress to observe the impact on NIS activity and energy charge. We used [18F]tetrafluoroborate positron emission tomography (PET) imaging to non-invasively image NIS activity in vivo.NIS activity was ablated by treating HEK293T cells in vitro, with the Na+/K+ ATPase inhibitor digoxin, confirming that radiotracer uptake was dependent on the sodium-potassium concentration gradient. NIS-mediated radiotracer uptake was significantly reduced (- 58.2%) following disruptions to ATP re-synthesis by combined glycolysis and oxidative phosphorylation inhibition in HEK293T cells and by oxidative phosphorylation inhibition (- 16.6%) in A549 cells in vitro. PET signal was significantly decreased (- 56.5%) within 90 min from the onset of treatment with IACS-010759, an oxidative phosphorylation inhibitor, in subcutaneous transgenic A549 tumours in vivo, showing that NIS could rapidly and sensitively detect energy stress non-invasively, before more widespread changes to phosphorylated AMP-activated protein kinase, phosphorylated pyruvate dehydrogenase, and GLUT1 were detectable.NIS acts as a rapid metabolic sensor for drugs that lead to ATP depletion. PET imaging of NIS could facilitate in vivo testing of treatments targeting energetic pathways, determine drug potency, and expedite metabolic drug development.© 2023. BioMed Central Ltd., part of Springer Nature.