Protein biomarkers for in vitro testing of embryotoxicity.
Protein biomarkers for in vitro testing of embryotoxicity.
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DOI：
10.1021/pr100514e
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发表时间：
2010-10
影响因子：
4.4
通讯作者：
K. Groebe;K. Hayess;Martina Klemm-Manns;G. Schwall;W. Wozny;M. Steemans;A. K. Peters;C. Sastri;P. Jaeckel;W. Stegmann;H. Zengerling;Rainer Schopf;S. Poznanović;T. Stummann;A. Seiler;H. Spielmann;A. Schrattenholz
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文献类型：
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作者：
K. Groebe;K. Hayess;Martina Klemm-Manns;G. Schwall;W. Wozny;M. Steemans;A. K. Peters;C. Sastri;P. Jaeckel;W. Stegmann;H. Zengerling;Rainer Schopf;S. Poznanović;T. Stummann;A. Seiler;H. Spielmann;A. Schrattenholz
在欧洲的REACH法规以及美国和日本的相关活动方面，化学工业中的危害和风险评估面临新的挑战，这需要开发新的体外模型，用于药物或化学相关效应的分子表征，以取代传统的动物试验。在欧洲FP 6生殖毒理学项目（www.reprotect.eu）的框架内，我们根据经验证的胚胎干细胞试验（EST）方案，从分化为收缩心肌细胞的小鼠胚胎干细胞中制备蛋白质样品，这些样品已暴露于由专家委员会从不同的体内胚胎毒性类别中选择的毒性物质。使用裂解物进行以下研究：（i）确定EST中的最佳剂量范围条件，所述最佳剂量范围条件适合于（ii）进行潜在分子途径的差异定量蛋白质组学研究，（iii）定义具有相似蛋白质组学响应模式的物质类别，（iv）将这些类别与传统的体内胚胎毒性类别联系起来，最终目标是鉴定用于胚胎毒性的新的替代蛋白生物标志物候选物。我们发现两类不同的有毒物质（地诺西、赭曲霉毒素A和硝苯地平vs β-氨基丙腈、甲氧氯普胺、琥珀酸多西拉敏和d-青霉胺）在其蛋白质组模式中具有明显的通路相关差异。最值得注意的是，观察到热休克蛋白β-1、Ras-GTdR激活蛋白SH 3-结构域结合蛋白、Ran结合蛋白5和钙网蛋白、二氢嘧啶酶样2（Ulip 2蛋白）对簇1和簇2物质的不同反应。另一方面，热休克蛋白8和Fscn 1蛋白被来自两个簇的所有化合物下调。
There are new challenges for hazard and risk assessment in the chemical industry with regard to REACH legislation in Europe and related activities in the U.S. and Japan, which require the development of novel in vitro models for the molecular characterization of drug- or chemical-related effects replacing conventional animal testing. In the frame of a European FP6 project on reproductive toxicology ( www.reprotect.eu ), we prepared protein samples from mouse embryonic stem cells differentiated into contracting cardiomyocytes according to the validated embryonic stem cell test (EST) protocol, which had been exposed to toxic substances selected by an expert committee from different in vivo categories of embryotoxicity. Lysates were used to carry out the following investigations: (i) identify optimal dose range conditions in the EST that are suitable for (ii) performing a differential quantitative proteomic study of underlying molecular pathways, (iii) define classes of substances with similar proteomic response patterns, (iv) relate these classes to the traditional in vivo categories of embryotoxicity with (v) the final goal to identify novel surrogate protein biomarker candidates for embryo toxicity. We found two distinct classes of toxic substances (Dinoseb, Ochratoxin-A, and Nitrofen vs β-aminoproprionitril, Metoclopramide, Doxylamine succinate, and d-penicillamine) with clear pathway-related differences in their proteomic patterns. Most notably, different responses to cluster 1 and cluster 2 substances were observed for Heat shock protein β-1, Ras-GTPase-activating protein SH3-domain binding protein, Ran binding protein 5, and Calreticulin, Dihydropyrimidinase-like 2 (Ulip2 protein). On the other hand, Heat shock protein 8 and Fscn1 protein were down-regulated by all compounds from both clusters.