Bioluminescent imaging of drug efflux at the blood-brain barrier mediated by the transporter ABCG2
Bioluminescent imaging of drug efflux at the blood-brain barrier mediated by the transporter ABCG2
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DOI：
10.1073/pnas.1312159110
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发表时间：
2013-12-17
影响因子：
11.1
通讯作者：
Hall, Matthew D.
中科院分区：
文献类型：
--
作者：
Bakhsheshian, Joshua;Wei, Bih-Rong;Hall, Matthew D.
ATP 结合盒 (ABC) 转运蛋白是一组跨膜蛋白，通过将化合物流出细胞器和细胞来维持化学稳态。除其他功能外，ABC 转运蛋白在通过血脑屏障 (BBB) 毛细血管内皮细胞流出异生物质来保护脑实质方面发挥着关键作用。它们还阻止治疗药物进入血脑屏障，从而限制其疗效。 ABCG2 是发挥这一作用的关键转运蛋白之一。尽管可以通过各种成像方式研究其他 ABC 转运蛋白，但不存在用于体内成像 ABCG2 功能的特异性探针。在这里，我们证明萤火虫荧光素酶的内源底物 D-荧光素是 ABCG2 的特异性底物。我们假设可以通过在大脑中表达萤火虫荧光素酶的转基因小鼠中使用生物发光成像来评估 BBB 处的 ABCG2 功能。联合使用 ABCG2 抑制剂 Ko143、吉非替尼和尼洛替尼（但不使用 ABCB1 抑制剂）后，小鼠大脑中的生物发光信号增加。这种在 BBB 处成像 ABCG2 功能的方法将有助于了解该转运蛋白的功能和药代动力学抑制。
ATP-binding cassette (ABC) transporters are a group of transmembrane proteins that maintain chemical homeostasis through efflux of compounds out of organelles and cells. Among other functions, ABC transporters play a key role in protecting the brain parenchyma by efflux of xenobiotics from capillary endothelial cells at the blood-brain barrier (BBB). They also prevent the entry of therapeutic drugs at the BBB, thereby limiting their efficacy. One of the key transporters playing this role is ABCG2. Although other ABC transporters can be studied through various imaging modalities, no specific probe exists for imaging ABCG2 function in vivo. Here we show that D-luciferin, the endogenous substrate of firefly luciferase, is a specific substrate for ABCG2. We hypothesized that ABCG2 function at the BBB could be evaluated by using bioluminescence imaging in transgenic mice expressing firefly luciferase in the brain. Bioluminescence signal in the brain of mice increased with co-administration of the ABCG2 inhibitors Ko143, gefitinib, and nilotinib, but not an ABCB1 inhibitor. This method for imaging ABCG2 function at the BBB will facilitate understanding of the function and pharmacokinetic inhibition of this transporter.