Is the human model RPTEC/TERT1 a relevant model for assessing renal drug efflux?
Is the human model RPTEC/TERT1 a relevant model for assessing renal drug efflux?
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人类模型 RPTEC/TERT1 是否是评估肾脏药物流出的相关模型？
DOI：
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发表时间：
2020
影响因子：
2.9
通讯作者：
X. Delavenne
中科院分区：
文献类型：
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作者：
S. Saib;S. Hodin;Zhiguo Hé;O. Delézay;X. Delavenne
由于存在许多膜转运蛋白如ABC转运蛋白，活性肾小管分泌在药物的肾排泄中起主要作用。这些蛋白质促进药物转移到尿液中，可能是药代动力学变异性的来源。迄今为止，已经提出了几种人近端小管体外模型，但很少有人能用于预测药物的肾外排。本研究的目的是确定人模型RPTEC/TERT 1是否符合评估肾脏药物转运的良好模型的所有预期标准。首先，研究了体外屏障特性。然后，与MDCK模型相比，通过免疫荧光和液相色谱-高分辨率质谱（LC-HRMS）相对定量评估了几种ABC转运蛋白的表达。最后，进行双向转运研究以评估转运蛋白的功能和模型区分几种药物的能力。RPTEC/TERT 1模型形成了一个紧密的结构（192 μ m.cm 2），这是通过细胞旁渗透性测定证实的。蛋白质组学分析和免疫荧光染色显示了几个ABC转运蛋白的表达。然后，在本研究中，仅通过阿哌沙班的主动外排证实了P-gp的功能性。此外，RPTEC/TERT 1模型提供了肾屏障的关键标准，并表达了几种ABC转运蛋白。然而，BCRP和MRP的功能尚未得到证实，需要进一步研究以验证该模型作为评估肾脏药物外排的体外模型的有效性。
Active tubular secretion plays a major role in renal excretion of drugs thanks to the presence of many membrane transporters such as ABC transporters. These proteins facilitate drug transfer into the urine and could be a source of pharmacokinetic variabilities. Up to now, several human in vitro models of proximal tubule have been proposed but few of them have been characterized for predicting drugs renal efflux. The aim of this study was to determine whether the human model RPTEC/TERT1 meets all the criteria expected of a good model to assess renal drug transport. First, in vitro barrier properties were investigated. Then, the expression of several ABC transporters was assessed by immunofluorescence and relative quantification by liquid chromatography—high‐resolution mass spectrometry (LC‐HRMS) in comparison to the MDCK model. Finally, bidirectional transport studies were performed to evaluate the functionality of transporters and the abilities of model to discriminate several drugs. The RPTEC/TERT1 model formed a tight structure (192 Ω.cm2) that was confirmed by paracellular permeability assays. Proteomic analysis and immunofluorescence staining showed the expression of several ABC transporters. Then, only the functionality of P‐gp was confirmed by the active efflux of apixaban in this study. In addition, the RPTEC/TERT1 model presents the key criteria of a renal barrier and expresses several ABC transporters. Nevertheless, the BCRP and MRP’s functionality was not confirmed and further investigations are required to valid this model as in vitro model for assessing renal drug efflux.