Pathology tissue-chromatin immunoprecipitation, coupled with high-throughput sequencing, allows the epigenetic profiling of patient samples
Pathology tissue-chromatin immunoprecipitation, coupled with high-throughput sequencing, allows the epigenetic profiling of patient samples
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DOI：
10.1073/pnas.1007647107
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发表时间：
2010-12-14
影响因子：
11.1
通讯作者：
Minucci, Saverio
中科院分区：
文献类型：
--
作者：
Fanelli, Mirco;Amatori, Stefano;Minucci, Saverio
DNA和组蛋白修饰模式的表观遗传改变在癌症发展中起着至关重要的作用。然而，患者样本的分析受到来自病理学档案的染色质结构研究的技术限制的阻碍，所述病理学档案通常由高度固定的石蜡包埋材料组成。在这里，我们提出了一种方法[病理组织-ChIP（PAT-ChIP）]，从石蜡包埋的患者样本中提取染色质并进行免疫沉淀，最多可达几年。在与经典ChIP的成对比较中，PAT-ChIP显示了几种组蛋白标记的结果的高度再现性和在药物治疗后检测染色质结构动态变化的相同能力。最后，我们表明PAT-ChIP可以与高通量测序（PAT-ChIP-Seq）结合，用于不同染色质修饰的全基因组分析。因此，PAT-ChIP代表了一种用于分析癌症和潜在其他疾病中表观遗传改变的通用程序和诊断工具。
Epigenetic alterations in the pattern of DNA and histone modifications play a crucial role in cancer development. Analysis of patient samples, however, is hampered by technical limitations in the study of chromatin structure from pathology archives that usually consist of heavily fixed, paraffin-embedded material. Here, we present a methodology [pathology tissue-ChIP (PAT-ChIP)] to extract and immunoprecipitate chromatin from paraffin-embedded patient samples up to several years old. In a pairwise comparison with canonical ChIP, PAT-ChIP showed a high reproducibility of results for several histone marks and an identical ability to detect dynamic changes in chromatin structure upon pharmacological treatment. Finally, we showed that PAT-ChIP can be coupled with high-throughput sequencing (PAT-ChIP-Seq) for the genome-wide analysis of distinct chromatin modifications. PAT-ChIP therefore represents a versatile procedure and diagnostic tool for the analysis of epigenetic alterations in cancer and potentially other diseases.