Advances in blood-brain barrier modeling in microphysiological systems highlight critical differences in opioid transport due to cortisol exposure
Advances in blood-brain barrier modeling in microphysiological systems highlight critical differences in opioid transport due to cortisol exposure
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DOI：
10.1186/s12987-020-00200-9
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发表时间：
2020-06-03
影响因子：
7.3
通讯作者：
Lippmann, Ethan S.
中科院分区：
文献类型：
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作者：
Brown, Jacquelyn A.;Faley, Shannon L.;Lippmann, Ethan S.
背景美国面临着一场涉及阿片类药物的全国性危机，目前每天有130多人死亡。为了对抗这种流行病，需要更好地了解阿片类药物如何渗透到中枢神经系统（CNS）以促进疼痛缓解，并可能导致成瘾和/或滥用。然而，动物模型是人体血脑屏障（BBB）转运和CNS药物渗透的不良预测因子，并且BBB和神经血管单元的许多传统2D细胞培养模型具有不足的屏障功能和弱或不适当的外排转运蛋白表达。在这里，我们试图更好地了解阿片类药物的转运机制，使用一个简化的微流体神经血管单元（NVU）模型组成的人脑微血管内皮细胞（BMEC）与星形胶质细胞共培养。方法将人原代和诱导多能干细胞（iPSC）衍生的BMEC纳入微流控NVU模型中，与我们以前的设计相比进行了几项技术改进。被动屏障功能通过不同大小的荧光葡聚糖的渗透性进行评估，P-糖蛋白功能通过在存在或不存在抑制剂的情况下的罗丹明渗透性进行评估;用荧光酶标仪进行定量。在存在或不存在P-糖蛋白抑制剂和皮质醇的情况下评估洛哌丁胺、吗啡和羟考酮的渗透性;采用质谱法进行定量。结果我们首先报告了对我们先前描述的使用原代人BMEC的微流体模型的技术和方法优化，这导致相对于Transwell模型的屏障形成加速，可变性降低和被动渗透性降低。然后，我们证明了洛哌丁胺、吗啡和羟考酮在含有源自人iPSC的BMEC的微流体NVU中的适当转运和流出。我们进一步证明，皮质醇可以改变洛哌丁胺和吗啡的渗透性在不同的方式。结论我们揭示了应激激素皮质醇在调节阿片类药物通过血脑屏障的转运中的新作用，这可能导致阿片类药物滥用或过量。我们更新的BBB模型是研究人员，临床医生和药物制造商了解阿片类药物进入CNS的机制的强大工具。
Background The United States faces a national crisis involving opioid medications, where currently more than 130 people die every day. To combat this epidemic, a better understanding is needed of how opioids penetrate into the central nervous system (CNS) to facilitate pain relief and, potentially, result in addiction and/or misuse. Animal models, however, are a poor predictor of blood-brain barrier (BBB) transport and CNS drug penetration in humans, and many traditional 2D cell culture models of the BBB and neurovascular unit have inadequate barrier function and weak or inappropriate efflux transporter expression. Here, we sought to better understand opioid transport mechanisms using a simplified microfluidic neurovascular unit (NVU) model consisting of human brain microvascular endothelial cells (BMECs) co-cultured with astrocytes. Methods Human primary and induced pluripotent stem cell (iPSC)-derived BMECs were incorporated into a microfluidic NVU model with several technical improvements over our previous design. Passive barrier function was assessed by permeability of fluorescent dextrans with varying sizes, and P-glycoprotein function was assessed by rhodamine permeability in the presence or absence of inhibitors; quantification was performed with a fluorescent plate reader. Loperamide, morphine, and oxycodone permeability was assessed in the presence or absence of P-glycoprotein inhibitors and cortisol; quantification was performed with mass spectrometry. Results We first report technical and methodological optimizations to our previously described microfluidic model using primary human BMECs, which results in accelerated barrier formation, decreased variability, and reduced passive permeability relative to Transwell models. We then demonstrate proper transport and efflux of loperamide, morphine, and oxycodone in the microfluidic NVU containing BMECs derived from human iPSCs. We further demonstrate that cortisol can alter permeability of loperamide and morphine in a divergent manner. Conclusions We reveal a novel role for the stress hormone cortisol in modulating the transport of opioids across the BBB, which could contribute to their abuse or overdose. Our updated BBB model represents a powerful tool available to researchers, clinicians, and drug manufacturers for understanding the mechanisms by which opioids access the CNS.