Establishment and assessment of a new human embryonic stem cell-based biomarker assay for developmental toxicity screening.
Establishment and assessment of a new human embryonic stem cell-based biomarker assay for developmental toxicity screening.
复制标题
DOI：
10.1002/bdrb.21078
复制
发表时间：
2013-08-01
期刊：
影响因子：
--
通讯作者：
Kirchner, Fred R
中科院分区：
文献类型：
--
作者：
Palmer, Jessica A;Smith, Alan M;Kirchner, Fred R
开发了利用人胚胎干（hES）细胞的基于代谢生物标志物的体外测定，以鉴定以指示致畸性的方式干扰细胞代谢的测试化合物的浓度。本试验旨在帮助早期发现阶段检测潜在的人类发育毒物。在本研究中，使用来自hES细胞培养基的代谢组学数据评估用于开发快速体外致畸性试验的潜在生物标志物。用已知的人类致畸性的药物处理hES细胞，其浓度相当于其公开的人类峰值治疗血浆浓度。两种代谢物生物标志物（鸟氨酸和胱氨酸）被确定为发育毒性的指标。然后使用9点剂量-反应曲线开发了使用这些代谢物的基于靶向清除的生物标志物测定，沿着细胞毒性终点。使用一组单独的供试化合物评价新测定法的预测性。为了说明该试验如何应用于具有未知发育毒性潜力的化合物，对另外10种化合物进行了评价，这些化合物没有妊娠期间人体暴露的数据，但在动物发育毒性研究中显示出阳性结果。新的检测方法以77%的准确度（57%的灵敏度，100%的特异性）鉴定了测试集中的潜在发育毒物。该试验与现有体内模型具有高度一致性（≥75%），表明新试验可预测新化合物的发育毒性潜力（作为发现阶段试验的一部分），并提供所需体内试验可能结果的信号。
A metabolic biomarker-based in vitro assay utilizing human embryonic stem (hES) cells was developed to identify the concentration of test compounds that perturbs cellular metabolism in a manner indicative of teratogenicity. This assay is designed to aid the early discovery-phase detection of potential human developmental toxicants. In this study, metabolomic data from hES cell culture media were used to assess potential biomarkers for development of a rapid in vitro teratogenicity assay. hES cells were treated with pharmaceuticals of known human teratogenicity at a concentration equivalent to their published human peak therapeutic plasma concentration. Two metabolite biomarkers (ornithine and cystine) were identified as indicators of developmental toxicity. A targeted exposure-based biomarker assay using these metabolites, along with a cytotoxicity endpoint, was then developed using a 9-point dose-response curve. The predictivity of the new assay was evaluated using a separate set of test compounds. To illustrate how the assay could be applied to compounds of unknown potential for developmental toxicity, an additional 10 compounds were evaluated that do not have data on human exposure during pregnancy, but have shown positive results in animal developmental toxicity studies. The new assay identified the potential developmental toxicants in the test set with 77% accuracy (57% sensitivity, 100% specificity). The assay had a high concordance (≥75%) with existing in vivo models, demonstrating that the new assay can predict the developmental toxicity potential of new compounds as part of discovery phase testing and provide a signal as to the likely outcome of required in vivo tests.