Progress in researches on in vitro alternative model for nephrotoxicity evaluation
-
摘要: 随着《21世纪毒性测试:愿景与策略》的提出,毒性测试的重点开始从整体动物实验转向基于人类细胞或细胞组分等替代方法的测试策略。由于肾毒性物质的靶器官选择性,且药物诱导的肾损伤是新药开发时需解决的重要问题,故需良好的体外替代模型来评价包括药物在内的外源性化合物的肾脏毒性。然而,现有的体外模型由于缺少肾小管上皮细胞在体内的形态及功能,难以预测外源性化合物的肾脏毒性。肾体外替代模型的细胞来源、特点、培养条件以及检测终点是在建立体外替代模型时重点考虑的问题。多功能干细胞诱导分化肾细胞的出现、3D培养及肾脏芯片技术的发展、组学技术、高内涵筛选的应用为体外模型的建立及模型预测结果从体外到体内的外推提供了新思路。Abstract: With the introduction of Toxicity Testing in the 21st Century: A Vision and Strategy, the focus of toxicity testing has shifted from animal experiments to in vitro models using human resource cells or cellular components. As kidney is a major target for drug-induced toxicity and the drug-induced toxicity remains a major problem in developing new drugs, a predictive in vitro model is urgently needed to evaluate the renal toxicity of exogenous compounds. However, current in vitro cellular models poorly replicate both the morphology and the function of kidney tubules and therefore fail to demonstrate injury responses to that would be nephrotoxic in vivo. The resource and characteristics of cellular models, cell culture conditions, and readouts of injury are important in establishing an in vitro model. The development of differentiation of pluripotent stem cells into multiple renal cell types, 3 dimensional culture systems and kidney-on-a-chip technology, omics technology and high-content screening have opened a range of potential new platforms for evaluating compounds nephrotoxicity and promoted in vitro to in vivo extrapolation. This study summarizes the latest advances in in vitro nephrotoxicity assessment models.
-
表 1 与急性肾损伤相关的体内及体外生物标志物
生物标志物 标志性损伤 部位特异性 参考文献 FDA及EMA认证的生物标志物 肾损伤因子 毒性、缺血引起AKI/早期急性肾小管损伤 近端小管 [32 – 34] 丛生蛋白 毒性、缺血引起AKI、单侧输尿管梗阻 近端小管及远端小管 [32 – 33] 三叶因子 – 3 与肾组织病理学损伤相关 近端小管 [32 – 33] β2微球蛋白 毒性、缺血引起AKI、肾脏移植 肾小球及近端小管 [32] 血清半胱氨酸蛋白酶抑制剂C 肾小球滤过功能、毒性、缺血引起AKI 肾小球及近端小管 [32 – 33] 尿总蛋白 慢性肾损伤(CKD)、肾小球滤过功能或肾小管重吸收功能受损 肾小球及肾小管 [32] 白蛋白 毒性、缺血引起AKI 肾小球及近端小管 [32 – 33] 肾乳头抗原 – 1 集合管损伤 集合管 [32] 其他肾脏相关生物标志物 γ – 谷氨酰转肽酶 肾小管上皮刷状缘受损 肾小管上皮刷状缘 [35] N – 乙酰 – β – D – 葡萄糖苷酶 毒性、缺血引起AKI、肾脏移植、慢性肾小球疾病 近端小管溶酶体 [36] 骨桥蛋白 毒性、缺血引起AKI、单侧输尿管梗阻 无部位特异性 [37] 钙结合蛋白 远端小管及集合管功能 无部位特异性 [37 – 38] 基质金属蛋白酶抑制剂 – 1 肾间质纤维化 无部位特异性 [38] 中性粒细胞明胶酶相关载脂蛋白 毒性、缺血引起AKI/与炎症相关 近端小管及远端小管 [38 – 39] 白介素 – 18 炎症反应/毒性、缺血引起AKI 近端小管 [21] 白介素 – 6 炎症反应 能够在近端小管细胞中表达 [18, 21] 白介素 – 8 炎症反应 能够在近端小管细胞中表达 [18, 21] 血红素加氧酶 – 1 氧化应激、炎症反应 近端小管、远端小管、集合管及髓袢上皮细胞 [17] -
[1] Choudhury D, Ahmed Z. Drug-associated renal dysfunction and injury[J]. Nature Clinical Practice Nephrology, 2006, 2(2): 80 – 91. doi: 10.1038/ncpneph0076 [2] Gonsalez SR, Cortês AL, da Silva RC, et al. Acute kidney injury overview: from basic findings to new prevention and therapy strategies[J]. Pharmacology and Therapeutics, 2019, 200: 1 – 12. doi: 10.1016/j.pharmthera.2019.04.001 [3] Mehta RL, Awdishu L, Davenport A, et al. Phenotype standardization for drug-induced kidney disease[J]. Kidney International, 2015, 88(2): 226 – 234. doi: 10.1038/ki.2015.115 [4] Scotcher D, Jones C, Posada M, et al. Key to opening kidney for in vitro – in vivo extrapolation entrance in health and disease: Part Ⅱ: mechanistic models and in vitro – in vivo extrapolation[J]. The AAPS Journal, 2016, 18(5): 1082 – 1094. doi: 10.1208/s12248-016-9959-1 [5] Soo JYC, Jansen J, Masereeuw R, et al. Advances in predictive in vitro models of drug-induced nephrotoxicity[J]. Nature Reviews Nephrology, 2018, 14(6): 378 – 393. doi: 10.1038/s41581-018-0003-9 [6] Aleksa K, Halachmi N, Ito S, et al. A tubule cell model for ifosfamide nephrotoxicity[J]. Canadian Journal of Physiology and Pharmacology, 2005, 83(6): 499 – 508. doi: 10.1139/y05-036 [7] Alvarez-Barrientos A, O'Connor JE, Castillo RN, et al. Use of flow cytometry and confocal microscopy techniques to investigate early CdCl2-induced nephrotoxicity in vitro[J]. Toxicology in Vitro, 2001, 15(4/5): 407 – 412. [8] Schinkel AH, Mayer U, Wagenaar E, et al. Normal viability and altered pharmacokinetics in mice lacking mdr1-type (drug-transporting) P-glycoproteins[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(8): 4028 – 4033. doi: 10.1073/pnas.94.8.4028 [9] Ryan MJ, Johnson G, Kirk J, et al. HK-2: an immortalized proximal tubule epithelial cell line from normal adult human kidney[J]. Kidney International, 1994, 45(1): 48 – 57. doi: 10.1038/ki.1994.6 [10] Kim D, Garrett SH, Sens MA, et al. Metallothionein isoform 3 and proximal tubule vectorial active transport[J]. Kidney International, 2002, 61(2): 464 – 472. doi: 10.1046/j.1523-1755.2002.00153.x [11] Jenkinson SE, Chung GW, Van Loon E, et al. The limitations of renal epithelial cell line HK-2 as a model of drug transporter expression and function in the proximal tubule[J]. Pflugers Archiv- European Journal of Physiology, 2012, 464(6): 601 – 611. doi: 10.1007/s00424-012-1163-2 [12] Wieser M, Stadler G, Jennings P, et al. hTERT alone immortalizes epithelial cells of renal proximal tubules without changing their functional characteristics[J]. American Journal of Physiology- Renal Physiology, 2008, 295(5): F1365 – F1375. doi: 10.1152/ajprenal.90405.2008 [13] Wilmes A, Limonciel A, Aschauer L, et al. Application of integrated transcriptomic, proteomic and metabolomic profiling for the delineation of mechanisms of drug induced cell stress[J]. Journal of Proteomics, 2013, 79: 180 – 194. doi: 10.1016/j.jprot.2012.11.022 [14] Aschauer L, Limonciel A, Wilmes A, et al. Application of RPTEC/TERT1 cells for investigation of repeat dose nephrotoxi-city: a transcriptomic study[J]. Toxicology in Vitro, 2015, 30(1): 106 – 116. doi: 10.1016/j.tiv.2014.10.005 [15] Mueller SO, Dekant W, Jennings P, et al. Comprehensive summary – Predict – IV: a systems toxicology approach to improve pharmaceu-tical drug safety testing[J]. Toxicology in Vitro, 2015, 30(1): 4 – 6. doi: 10.1016/j.tiv.2014.09.016 [16] Wilmer MJ, Saleem MA, Masereeuw R, et al. Novel conditionally immortalized human proximal tubule cell line expressing functional influx and efflux transporters[J]. Cell and Tissue Research, 2010, 339(2): 449 – 457. doi: 10.1007/s00441-009-0882-y [17] Adler M, Ramm S, Hafner M, et al. A quantitative approach to screen for nephrotoxic compounds in vitro[J]. Journal of the American Society of Nephrology, 2016, 27(4): 1015 – 1028. doi: 10.1681/ASN.2015010060 [18] LI Y, Oo ZY, Chang SY, et al. An in vitro method for the prediction of renal proximal tubular toxicity in humans[J]. Toxicology Research, 2013, 2(5): 352 – 365. doi: 10.1039/c3tx50042j [19] Davies J. Engineered renal tissue as a potential platform for pharmacokinetic and nephrotoxicity testing[J]. Drug Discovery Today, 2014, 19(6): 725 – 729. doi: 10.1016/j.drudis.2013.10.023 [20] Narayanan K, Schumacher KM, Tasnim F, et al. Human embryonic stem cells differentiate into functional renal proximal tubular-like cells[J]. Kidney International, 2013, 83(4): 593 – 603. doi: 10.1038/ki.2012.442 [21] LI Y, Kandasamy K, Chuah JKC, et al. Identification of nephrotoxic compounds with embryonic stem-cell-derived human renal proximal tubular-like cells[J]. Molecular Pharmaceutics, 2014, 11(7): 1982 – 1990. doi: 10.1021/mp400637s [22] Kandasamy K, Chuah JKC, Su R, et al. Prediction of drug-induced nephrotoxicity and injury mechanisms with human induced pluripotent stem cell-derived cells and machine learning methods[J]. Scientific Reports, 2015, 5: 12337. doi: 10.1038/srep12337 [23] Astashkina AI, Mann BK, Prestwich GD, et al. Comparing predictive drug nephrotoxicity biomarkers in kidney 3-D primary organoid culture and immortalized cell lines[J]. Biomaterials, 2012, 33(18): 4712 – 4721. doi: 10.1016/j.biomaterials.2012.03.001 [24] Wilmer MJ, Ng CP, Lanz HL, et al. Kidney-on-a-chip technology for drug-induced nephrotoxicity screening[J]. Trends in Biotechnology, 2016, 34(2): 156 – 170. doi: 10.1016/j.tibtech.2015.11.001 [25] Jang KJ, Mehr AP, Hamilton GA, et al. Human kidney proximal tubule-on-a-chip for drug transport and nephrotoxicity assessment[J]. Integrative Biology, 2013, 5(9): 1119 – 1129. doi: 10.1039/c3ib40049b [26] Trietsch SJ, Israëls GD, Joore J, et al. Microfluidic titer plate for stratified 3D cell culture[J]. Lab on a Chip, 2013, 13(18): 3548 – 3554. doi: 10.1039/c3lc50210d [27] Lever JE. Inducers of dome formation in epithelial cell cultures including agents that cause differentiation[M]//Taub M. Tissue Culture of Epithelial Cells. Boston: Springer, 1985: 3 – 22. [28] Nieskens TTG, Peters JGP, Schreurs MJ, et al. A human renal proximal tubule cell line with stable organic anion transporter 1 and 3 expression predictive for antiviral-induced toxicity[J]. The AAPS Journal, 2016, 18(2): 465 – 475. doi: 10.1208/s12248-016-9871-8 [29] Hagos Y, Wolff NA. Assessment of the role of renal organic anion transporters in drug-induced nephrotoxicity[J]. Toxins, 2010, 2(8): 2055 – 2082. doi: 10.3390/toxins2082055 [30] Ciarimboli G. Role of organic cation transporters in drug-induced toxicity[J]. Expert Opinion on Drug Metabolism and Toxicology, 2011, 7(2): 159 – 174. doi: 10.1517/17425255.2011.547474 [31] Hori Y, Aoki N, Kuwahara S, et al. Megalin blockade with cilastatin suppresses drug-induced nephrotoxicity[J]. Journal of the American Society of Nephrology, 2017, 28(6): 1783 – 1791. doi: 10.1681/ASN.2016060606 [32] Dieterle F, Sistare F, Goodsaid F, et al. Renal biomarker qualification submission: a dialog between the FDA-EMEA and Predictive Safety Testing Consortium[J]. Nature Biotechnology, 2010, 28(5): 455 – 462. doi: 10.1038/nbt.1625 [33] Qiu X, Zhou XB, Miao YF, et al. An in vitro method for nephrotoxicity evaluation using HK-2 human kidney epithelial cells combined with biomarkers of nephrotoxicity[J]. Toxicology Research, 2018, 7(6): 1205 – 1213. doi: 10.1039/C8TX00095F [34] Luo QH, Chen ML, Chen ZL, et al. Evaluation of KIM-1 and NGAL as early indicators for assessment of gentamycin-induced nephrotoxicity in vivo and in vitro[J]. Kidney and Blood Pressure Research, 2016, 41(6): 911 – 918. doi: 10.1159/000452592 [35] Westhuyzen J, Endre ZH, Reece G, et al. Measurement of tubular enzymuria facilitates early detection of acute renal impairment in the intensive care unit[J]. Nephrology Dialysis Transplantation, 2003, 18(3): 543 – 551. doi: 10.1093/ndt/18.3.543 [36] Vaidya VS, Ozer JS, Dieterle F, et al. Kidney injury molecule-1 outperforms traditional biomarkers of kidney injury in preclinical biomarker qualification studies[J]. Nature Biotechnology, 2010, 28(5): 478 – 485. doi: 10.1038/nbt.1623 [37] Hoffmann D, Fuchs TC, Henzler T, et al. Evaluation of a urinary kidney biomarker panel in rat models of acute and subchronic nephrotoxicity[J]. Toxicology, 2010, 277(1/3): 49 – 58. [38] Sohn SJ, Kim SY, Kim HS, et al. In vitro evaluation of biomarkers for cisplatin-induced nephrotoxicity using HK-2 human kidney epithelial cells[J]. Toxicology Letters, 2013, 217(3): 235 – 242. doi: 10.1016/j.toxlet.2012.12.015 [39] Wasilewska A, Zoch-Zwierz W, Taranta-Janusz K, et al. Neutrophil gelatinase-associated lipocalin (NGAL): a new marker of cyclosporine nephrotoxicity?[J]. Pediatric Nephrology, 2010, 25(5): 889 – 897. doi: 10.1007/s00467-009-1397-1 [40] Saikumar J, Hoffmann D, Kim TM, et al. Expression, circulation, and excretion profile of microRNA-21, -155, and -18a following acute kidney injury[J]. Toxicological Sciences, 2012, 129(2): 256 – 267. doi: 10.1093/toxsci/kfs210 [41] Sonoda H, Lee BR, Park KH, et al. miRNA profiling of urinary exosomes to assess the progression of acute kidney injury[J]. Scientific Reports, 2019, 9(1): 4692. doi: 10.1038/s41598-019-40747-8 [42] Su R, Xiong SJ, Zink D, et al. High-throughput imaging-based nephrotoxicity prediction for xenobiotics with diverse chemical structures[J]. Archives of Toxicology, 2016, 90(11): 2793 – 2808. doi: 10.1007/s00204-015-1638-y [43] Sjögren AK, Breitholtz K, Ahlberg E, et al. A novel multi-parametric high content screening assay in ciPTEC-OAT1 to predict drug-induced nephrotoxicity during drug discovery[J]. Archives of Toxicology, 2018, 92(10): 3175 – 3190. doi: 10.1007/s00204-018-2284-y
点击查看大图
表(1)
计量
- 文章访问数: 2949
- HTML全文浏览量: 1393
- PDF下载量: 79
- 被引次数: 0