Implementation of a Human Renal Proximal Tubule on a Chip for Nephrotoxicity and Drug Interaction Studies
Implementation of a Human Renal Proximal Tubule on a Chip for Nephrotoxicity and Drug Interaction Studies
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DOI：
10.1016/j.xphs.2021.01.028
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发表时间：
2021-03-23
影响因子：
3.8
通讯作者：
Suter-Dick, Laura
中科院分区：
文献类型：
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作者：
Vormann, Marianne K.;Vriend, Jelle;Suter-Dick, Laura
近端小管上皮细胞（PTEC）对药物性肾损伤（DIKI）敏感。基于细胞的二维（2D）体外PTEC模型通常是DIKI的不良预测因子，可能是由于缺乏生理结构和流动。在这里，我们评估了一个高通量，3D微流体平台（Nephroscreen）用于药物开发中DIKI的检测。该系统建立了四个模型肾毒性药物（顺铂，替诺福韦，妥布霉素和环孢菌素A），并与8个药物化合物进行了测试。测量的参数包括细胞活力、乳酸脱氢酶（LDH）和N-乙酰基-β-D-氨基葡萄糖苷酶（NAG）的释放、屏障完整性、特异性miRNA的释放和毒性标志物的基因表达。还测定了P-gp和MRP 2/4的药物-转运蛋白相互作用。DIKI最具预测性的读数是细胞活力、LDH和miRNA释放的组合。总之，Nephroscreen以稳健的方式检测DIKI，与自动移液兼容，证明适合长期实验，并且易于在实验室之间转移。该概念验证研究证明了Nephroscreen用于检测DIKI和药物-转运蛋白相互作用的可用性和重现性。它代表了一个有价值的工具，以取代动物实验和支持3R（减少，优化和取代动物实验）。(C)2021作者爱思唯尔公司出版。美国药学协会（American Pharmacopoeia Association）（R）
Proximal tubule epithelial cells (PTEC) are susceptible to drug-induced kidney injury (DIKI). Cell-based, two-dimensional (2D) in vitro PTEC models are often poor predictors of DIKI, probably due to the lack of physiological architecture and flow. Here, we assessed a high throughput, 3D microfluidic platform (Nephroscreen) for the detection of DIKI in pharmaceutical development. This system was established with four model nephrotoxic drugs (cisplatin, tenofovir, tobramycin and cyclosporin A) and tested with eight pharmaceutical compounds. Measured parameters included cell viability, release of lactate dehydrogenase (LDH) and N-acetyl-beta-D-glucosaminidase (NAG), barrier integrity, release of specific miRNAs, and gene expression of toxicity markers. Drug-transporter interactions for P-gp and MRP2/4 were also determined. The most predictive read outs for DIKI were a combination of cell viability, LDH and miRNA release. In conclusion, Nephroscreen detected DIKI in a robust manner, is compatible with automated pipetting, proved to be amenable to long-term experiments, and was easily transferred between laboratories. This proof-of-concept-study demonstrated the usability and reproducibility of Nephroscreen for the detection of DIKI and drug-transporter interactions. Nephroscreen it represents a valuable tool towards replacing animal testing and supporting the 3Rs (Reduce, Refine and Replace animal experimentation). (C) 2021 The Authors. Published by Elsevier Inc. on behalf of the American Pharmacists Association (R).