Pathology tissue-chromatin immunoprecipitation, coupled with high-throughput sequencing, allows the epigenetic profiling of patient samples

被引:48
作者
Fanelli, Mirco [1 ]
Amatori, Stefano [1 ,2 ]
Barozzi, Iros [2 ]
Soncini, Matias [2 ]
Dal Zuffo, Roberto [2 ]
Bucci, Gabriele [2 ]
Capra, Maria [2 ]
Quarto, Micaela [2 ,3 ]
Dellino, Gaetano Ivan [2 ,4 ]
Mercurio, Ciro [2 ,5 ]
Alcalay, Myriam [2 ,4 ]
Viale, Giuseppe [6 ]
Pelicci, Pier Giuseppe [2 ,4 ]
Minucci, Saverio [2 ,7 ]
机构
[1] Univ Urbino Carlo Bo, Dept Biomol Sci, I-61032 Fano, Italy
[2] European Inst Oncol, Dept Expt Oncol, I-20139 Milan, Italy
[3] Inst Mol Oncol Fdn, Fdn Italiana Ric Canc, IFOM, I-20100 Milan, Italy
[4] Univ Milan, Dept Med Surg & Dent, I-20124 Milan, Italy
[5] DAC Genextra Grp, I-20100 Milan, Italy
[6] European Inst Oncol, Div Pathol, I-20141 Milan, Italy
[7] Univ Milan, Dept Biomol Sci & Biotechnol, I-20100 Milan, Italy
关键词
EMBRYONIC STEM-CELLS; METHYLATION STATES; GENOME; EXPRESSION; PLASTICITY; PATHWAYS; CANCER; CHIP;
D O I
10.1073/pnas.1007647107
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];
摘要
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.
引用
收藏
页码:21535 / 21540
页数:6
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