DNA methylation profiling of myelodysplastic syndrome hematopoietic progenitor cells during in vitro lineage-specific differentiation
DNA methylation profiling of myelodysplastic syndrome hematopoietic progenitor cells during in vitro lineage-specific differentiation
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DOI：
10.1016/j.exphem.2007.01.054
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发表时间：
2007-05-01
影响因子：
2.6
通讯作者：
Hofmann, Wolf-Karsten
中科院分区：
文献类型：
--
作者：
Hopfer, Olaf;Komor, Martina;Hofmann, Wolf-Karsten
不受调控的表观遗传机制可能参与骨髓增生异常综合征（mds）的发病机制。在MDS造血过程中，哪些基因被异常启动子甲基化沉默，尚未得到相应的研究。利用人类造血的体外分化模型，我们从13名MDS患者和7名健康供体中生成红细胞、血栓和颗粒的明确分化阶段（第0天、第4天、第7天、第11天）。通过甲基化特异性聚合酶链反应对参与细胞周期控制（p14、p15、p16、CHK2）、DNA修复（hMLH1）、凋亡（p73、survivin、DAPK）和分化（RARb、WT1）的关键调控基因进行启动子甲基化分析。通过微阵列（Affymetrix, HG-U133A）分析相应基因的表达。我们提供的证据表明，mds中p16、survivin、CHK2和WT1受到启动子超甲基化的影响，显示出选择性的国际预后评分系统风险关联。研究表明，在特定的造血谱系和分化阶段，survivin、CHK2和WT1存在甲基化相关mRNA下调。我们发现，在红细胞生成过程中，甲基化样本中的survivin mRNA表达受到抑制，而在所有MDS风险类型的粒细胞生成过程中，WT1和CHK2甲基化相关的mRNA表达减少。我们的数据表明，MDS造血前体细胞中survivin、CHK2和WT1的谱系特异性甲基化相关基因沉默可能有助于MDS特异性表型。(c) 2007年国际实验血液学学会。Elsevier Inc.出版。
Deregulated epigenetic mechanisms are likely involved in the pathogenesis of myelodysplastic syndromes (MDSs). Which genes are silenced by aberrant promotor methylation during MDS hematopoiesis has not been equivalently investigated. Using an in vitro differentiation model of human hematopoiesis, we generated defined differentiation stages (day 0, day 4, day 7, day 11) of erythro-, thrombo- and granulopoiesis from 13 MDS patients and seven healthy donors. Promotor methylation analysis of key regulatory genes involved in cell cycle control (p14, p15, p16, CHK2), DNA repair (hMLH1), apoptosis (p73, survivin, DAPK), and differentiation (RARb, WT1) was performed by methylation-specific polymerase chain reaction. Corresponding gene expression was analyzed by microarray (Affymetrix, HG-U133A). We provide evidence that p16, survivin, CHK2, and WT1 are affected by promotor hypermethylation in MDSs displaying a selective International Prognostic Scoring System risk association. A methylation-associated mRNA downregulation for specific hematopoietic lineages and differentiation stages is demonstrated for survivin, CHK2, and WT1. We identified a suppressed survivin mRNA expression in methylated samples during erythropoiesis, whereas WT1 and CHK2 methylation-related reduction of mRNA expression was found during granulopoiesis in all MDS risk types. Our data suggest that lineage-specific methylation-associated gene silencing of survivin, CHK2, and WT1 in MDS hematopoietic precursor cells may contribute to the MDS-specific phenotype. (c) 2007 International Society for Experimental Hematology. Published by Elsevier Inc.