Evaluation of biomarkers for in vitro prediction of drug-induced nephrotoxicity: comparison of HK-2, immortalized human proximal tubule epithelial, and primary cultures of human proximal tubular cells.
Evaluation of biomarkers for in vitro prediction of drug-induced nephrotoxicity: comparison of HK-2, immortalized human proximal tubule epithelial, and primary cultures of human proximal tubular cells.
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DOI：
10.1002/prp2.148
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发表时间：
2015-06
影响因子：
2.6
通讯作者：
Cooper, Matthew A
中科院分区：
文献类型：
--
作者：
Huang, Johnny X;Kaeslin, Geraldine;Ranall, Max V;Blaskovich, Mark A;Becker, Bernd;Butler, Mark S;Little, Melissa H;Lash, Lawrence H;Cooper, Matthew A
人们付出了大量努力来识别体内生物标志物，这些生物标志物可用于监测药物引起的肾脏损伤，并在发生重大损害之前识别损伤。肾损伤分子 1 (KIM-1)、中性粒细胞明胶酶相关脂质运载蛋白 (NGAL) 和人巨噬细胞集落刺激因子 (M-CSF) 已被验证为预测急性和慢性肾损伤和疾病的尿液和血浆临床生物标志物。预测肾毒性的高通量体外测定的类似验证可能在药物发现先导化合物优化的早期实施，以减少药物开发后期的损耗。为了评估这些已知的体内生物标志物在体外筛选药物引起的肾毒性的潜力，我们选择了一组肾毒性药物，并检查了它们对永生化（HK-2）和原代（市售和新鲜内部生产的）人肾近曲小管上皮细胞中肾毒性生物标志物过度表达的影响。将传统的细胞毒性与使用 ELISA 和实时定量逆转录 PCR 评估的 KIM-1、NGAL 和 M-CSF 的表达水平进行对比。传统的细胞毒性测定和使用 HK-2 细胞的生物标志物测定均不适用于预测肾毒性。然而，原代细胞中 KIM-1 和 NGAL 蛋白水平的增加与已知肾毒性化合物的剂量水平密切相关，而 M-CSF 蛋白和 mRNA 水平的相关性有限。这些结果表明，通过监测生物标志物蛋白水平来分析针对原代细胞的化合物可能具有作为药物引起的肾毒性的体外预测测定的潜力。
There has been intensive effort to identify in vivo biomarkers that can be used to monitor drug-induced kidney damage and identify injury before significant impairment occurs. Kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), and human macrophage colony stimulating factor (M-CSF) have been validated as urinary and plasma clinical biomarkers predictive of acute and chronic kidney injury and disease. Similar validation of a high throughput in vitro assay predictive of nephrotoxicity could potentially be implemented early in drug discovery lead optimization to reduce attrition at later stages of drug development. To assess these known in vivo biomarkers for their potential for in vitro screening of drug-induced nephrotoxicity, we selected a panel of nephrotoxic agents and examined their effects on the overexpression of nephrotoxicity biomarkers in immortalized (HK-2) and primary (commercially available and freshly in-house produced) human renal proximal tubule epithelial cells. Traditional cytotoxicity was contrasted with expression levels of KIM-1, NGAL, and M-CSF assessed using ELISA and real-time quantitative reverse transcription PCR. Traditional cytotoxicity assays and biomarker assays using HK-2 cells were both unsuitable for prediction of nephrotoxicity. However, increases in protein levels of KIM-1 and NGAL in primary cells were well correlated with dose levels of known nephrotoxic compounds, with limited correlation seen in M-CSF protein and mRNA levels. These results suggest that profiling compounds against primary cells with monitoring of biomarker protein levels may have potential as in vitro predictive assays of drug-induced nephrotoxicity.