Division of Molecular and Computational Toxicology, Chemistry and Pharmaceutical Sciences, AIMMS, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Edelweiss Connect GmbH, Technology Park Basel, Hochbergerstrasse 60C, 4057, Basel, Switzerland.
Cell Biol Toxicol. 2023 Aug;39(4):1773-1793. doi: 10.1007/s10565-022-09783-5. Epub 2022 Dec 31.
Transcriptomic analysis is a powerful method in the utilization of New Approach Methods (NAMs) for identifying mechanisms of toxicity and application to hazard characterization. With this regard, mapping toxicological events to time of exposure would be helpful to characterize early events. Here, we investigated time-dependent changes in gene expression levels in iPSC-derived renal proximal tubular-like cells (PTL) treated with five diverse compounds using TempO-Seq transcriptomics with the aims to evaluate the application of PTL for toxicity prediction and to report on temporal effects for the activation of cellular stress response pathways. PTL were treated with either 50 μM amiodarone, 10 μM sodium arsenate, 5 nM rotenone, or 300 nM tunicamycin over a temporal time course between 1 and 24 h. The TGFβ-type I receptor kinase inhibitor GW788388 (1 μM) was used as a negative control. Pathway analysis revealed the induction of key stress-response pathways, including Nrf2 oxidative stress response, unfolding protein response, and metal stress response. Early response genes per pathway were identified much earlier than 24 h and included HMOX1, ATF3, DDIT3, and several MT1 isotypes. GW788388 did not induce any genes within the stress response pathways above, but showed deregulation of genes involved in TGFβ inhibition, including downregulation of CYP24A1 and SERPINE1 and upregulation of WT1. This study highlights the application of iPSC-derived renal cells for prediction of cellular toxicity and sheds new light on the temporal and early effects of key genes that are involved in cellular stress response pathways.
转录组分析是利用新型方法（NAMs）识别毒性机制并应用于危害特征描述的有力方法。在这方面，将毒理学事件映射到暴露时间有助于描述早期事件。在这里，我们使用 TempO-Seq 转录组学研究了五种不同化合物处理的 iPSC 衍生的肾近端管状样细胞 (PTL) 中基因表达水平随时间的变化，目的是评估 PTL 在毒性预测中的应用，并报告细胞应激反应途径的时间效应。PTL 用 50 μM 胺碘酮、10 μM 砷酸钠、5 nM 鱼藤酮或 300 nM 衣霉素处理，时间范围为 1 至 24 小时。TGFβ 型 I 受体激酶抑制剂 GW788388（1 μM）用作阴性对照。通路分析显示关键应激反应途径的诱导，包括 Nrf2 氧化应激反应、 unfolded protein response 和金属应激反应。每个途径的早期反应基因比 24 小时更早被识别，包括 HMOX1、ATF3、DDIT3 和几种 MT1 同工型。GW788388 未诱导上述应激反应途径中的任何基因，但显示 TGFβ 抑制相关基因的失调，包括 CYP24A1 和 SERPINE1 的下调以及 WT1 的上调。这项研究强调了 iPSC 衍生的肾细胞在预测细胞毒性中的应用，并为参与细胞应激反应途径的关键基因的时间和早期效应提供了新的认识。
