Verhelst Sigrid, De Clerck Laura, Willems Sander, Van Puyvelde Bart, Daled Simon, Deforce Dieter, Dhaenens Maarten
ProGenTomics, Laboratory of Pharmaceutical Biotechnology, Ghent University, Ottergemsesteenweg 460, 9000, Ghent, Belgium.
MethodsX. 2020 Sep 5;7:101055. doi: 10.1016/j.mex.2020.101055. eCollection 2020.
Evidence of the involvement of epigenetics in pathologies such as cancer, diabetes, and neurodegeneration has increased global interest in epigenetic modifications. For nearly thirty years, it has been known that cancer cells exhibit abnormal DNA methylation patterns. In contrast, the large-scale analysis of histone post-translational modifications (hPTMs) has lagged behind because classically, histone modification analysis has relied on site specific antibody-based techniques. Mass spectrometry (MS) is a technique that holds the promise to picture the histone code comprehensively in a single experiment. Therefore, we developed an MS-based method that is capable of tracking all possible hPTMs in an untargeted approach. In this way, trends in single and combinatorial hPTMs can be reported and enable prediction of the epigenetic toxicity of compounds. Moreover, this method is based on the use of human cells to provide preliminary data, thereby omitting the need to sacrifice laboratory animals. Improving the workflow and the user-friendliness in order to become a high throughput, easily applicable, toxicological screening assay is an ongoing effort. Still, this novel toxicoepigenetic assay and the data it generates holds great potential for, among others, pharmaceutical industry, food science, clinical diagnostics and, environmental toxicity screening. •There is a growing interest in epigenetic modifications, and more specifically in histone post-translational modifications (hPTMs).•We describe an MS-based workflow that is capable of tracking all possible hPTMs in an untargeted approach that makes use of human cells.•Improving the workflow and the user-friendliness in order to become a high throughput, easily applicable, toxicological screening assay is an ongoing effort.
表观遗传学参与癌症、糖尿病和神经退行性变等病理过程的证据，增加了全球对表观遗传修饰的关注。近三十年来，人们已经知道癌细胞表现出异常的DNA甲基化模式。相比之下，组蛋白翻译后修饰（hPTMs）的大规模分析则滞后了，因为传统上，组蛋白修饰分析依赖于基于位点特异性抗体的技术。质谱（MS）技术有望在一次实验中全面描绘组蛋白密码。因此，我们开发了一种基于质谱的方法，能够以非靶向方式追踪所有可能的hPTMs。通过这种方式，可以报告单个和组合hPTMs的趋势，并能够预测化合物的表观遗传毒性。此外，该方法基于使用人类细胞来提供初步数据，从而无需牺牲实验动物。为了成为一种高通量、易于应用的毒理学筛选测定方法，正在努力改进工作流程和提高用户友好性。尽管如此，这种新型的毒理表观遗传学测定方法及其产生的数据在制药行业、食品科学、临床诊断和环境毒性筛选等领域具有巨大潜力。
• 人们对表观遗传修饰，尤其是组蛋白翻译后修饰（hPTMs）的兴趣日益浓厚。
• 我们描述了一种基于质谱（MS）的工作流程，该流程能够以非靶向方式利用人类细胞追踪所有可能的hPTMs。
• 为了成为一种高通量、易于应用的毒理学筛选测定方法，正在努力改进工作流程和提高用户友好性。
