Han Dali, Lu Xingyu, Shih Alan H, Nie Ji, You Qiancheng, Xu Meng Michelle, Melnick Ari M, Levine Ross L, He Chuan
Department of Chemistry, Department of Biochemistry and Molecular Biology, and Institute for Biophysical Dynamics, Howard Hughes Medical Institute, University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
Leukemia Service, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA; Human Oncology and Pathogenesis Program and Center for Epigenetics Research, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA.
Mol Cell. 2016 Aug 18;63(4):711-719. doi: 10.1016/j.molcel.2016.06.028. Epub 2016 Jul 28.
We present a highly sensitive and selective chemical labeling and capture approach for genome-wide profiling of 5-hydroxylmethylcytosine (5hmC) using DNA isolated from ∼1,000 cells (nano-hmC-Seal). Using this technology, we assessed 5hmC occupancy and dynamics across different stages of hematopoietic differentiation. Nano-hmC-Seal profiling of purified Tet2-mutant acute myeloid leukemia (AML) murine stem cells allowed us to identify leukemia-specific, differentially hydroxymethylated regions that harbor known and candidate disease-specific target genes with differential 5hmC peaks compared to normal stem cells. The change of 5hmC patterns in AML strongly correlates with differential gene expression, demonstrating the importance of dynamic alterations of 5hmC in regulating transcription in AML. Together, covalent 5hmC labeling offers an effective approach to study and detect DNA methylation dynamics in in vivo disease models and in limited clinical samples.
我们提出了一种高度灵敏且具选择性的化学标记与捕获方法，用于使用从约1000个细胞中分离出的DNA进行全基因组5-羟甲基胞嘧啶（5hmC）分析（纳米hmC-Seal）。利用该技术，我们评估了造血分化不同阶段的5hmC占据情况和动态变化。对纯化的Tet2突变急性髓系白血病（AML）小鼠干细胞进行纳米hmC-Seal分析，使我们能够识别出白血病特异性的、差异羟甲基化区域，这些区域含有与正常干细胞相比具有差异5hmC峰的已知和候选疾病特异性靶基因。AML中5hmC模式的变化与差异基因表达密切相关，表明5hmC动态改变在调节AML转录中的重要性。总之，共价5hmC标记为研究和检测体内疾病模型及有限临床样本中的DNA甲基化动态提供了一种有效方法。
