Assessing DNA methylation in the developing human intestinal epithelium: potential link to inflammatory bowel disease.
Assessing DNA methylation in the developing human intestinal epithelium: potential link to inflammatory bowel disease.
复制标题
评估人类肠道上皮的DNA甲基化：与炎症性肠病的潜在联系。
DOI：
10.1038/mi.2015.88
复制
发表时间：
2016-05
影响因子：
8
通讯作者：
Zilbauer M
中科院分区：
文献类型：
--
作者：
Kraiczy J;Nayak K;Ross A;Raine T;Mak TN;Gasparetto M;Cario E;Rakyan V;Heuschkel R;Zilbauer M
DNA甲基化是调控细胞发育和细胞类型特异性基因表达的主要表观遗传学机制之一。在这里，我们同时对来自人胎儿肠道的纯化的肠上皮细胞、健康的儿科活检组织和新诊断为炎症性肠病(IBD)的儿童进行了全基因组DNA甲基化和基因表达分析。结果用焦磷酸测序、实时定量聚合酶链式反应和免疫染色进行了验证。通过在肠道细胞系中采用体外试验来评估DNA甲基化变化对基因表达的功能影响。DNA甲基化分析可以识别214个基因，这些基因的表达是通过DNA甲基化来调节的，即调节的差异甲基化区域(RDMRS)。RDMRS的途径和功能分析表明，DNA甲基化在调节人类肠道上皮细胞的基因表达和功能发育中起着关键作用。此外，对新诊断为IBD的儿童的肠道上皮细胞进行的分析显示，基因组内DNA甲基化发生了变化，这些变化与肠道发育过程中经历甲基化变化的情况显著重叠。我们的研究为DNA甲基化在调节人类肠上皮功能成熟中的生理作用提供了新的见解。此外，我们还提供了将发育获得性DNA甲基化改变与儿童IBD的改变联系起来的数据。
DNA methylation is one of the major epigenetic mechanisms implicated in regulating cellular development and cell-type-specific gene expression. Here we performed simultaneous genome-wide DNA methylation and gene expression analysis on purified intestinal epithelial cells derived from human fetal gut, healthy pediatric biopsies, and children newly diagnosed with inflammatory bowel disease (IBD). Results were validated using pyrosequencing, real-time PCR, and immunostaining. The functional impact of DNA methylation changes on gene expression was assessed by employing in-vitro assays in intestinal cell lines. DNA methylation analyses allowed identification of 214 genes for which expression is regulated via DNA methylation, i.e. regulatory differentially methylated regions (rDMRs). Pathway and functional analysis of rDMRs suggested a critical role for DNA methylation in regulating gene expression and functional development of the human intestinal epithelium. Moreover, analysis performed on intestinal epithelium of children newly diagnosed with IBD revealed alterations in DNA methylation within genomic loci, which were found to overlap significantly with those undergoing methylation changes during intestinal development. Our study provides novel insights into the physiological role of DNA methylation in regulating functional maturation of the human intestinal epithelium. Moreover, we provide data linking developmentally acquired alterations in the DNA methylation profile to changes seen in pediatric IBD.