Glycome Diagnosis of Human Induced Pluripotent Stem Cells Using Lectin Microarray
Glycome Diagnosis of Human Induced Pluripotent Stem Cells Using Lectin Microarray
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DOI：
10.1074/jbc.m111.231274
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发表时间：
2011-06-10
影响因子：
4.8
通讯作者：
Asashima, Makoto
中科院分区：
文献类型：
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作者：
Tateno, Hiroaki;Toyota, Masashi;Asashima, Makoto
通过四种转录因子的异位表达，可以从不同的体细胞(SC)系产生诱导多能干细胞(IPSCs)。虽然这一过程已经被证明可以诱导基因和microRNA表达的全球变化，甚至表观遗传修饰，但在很大程度上仍不清楚这种转录因子诱导的重编程如何影响细胞上表达的总糖链。在这里，我们对来自五种不同干细胞的114种人类IPSCs进行了全面的糖聚糖分析，并使用高密度凝集素微阵列将它们的糖类与人类胚胎干细胞(ESCs；九种细胞类型)的糖类进行了比较。在凝集素芯片结果的非监督聚类分析中，未分化的IPSCs和ESCs被聚为一个大组。然而，它们明显地从分化的干细胞群中分离出来，而所有四种干细胞都具有明显不同的糖组分，这表明最初具有不同糖组分的干细胞在诱导多能性后获得了类似于ESCs的糖组分。统计筛选了38个区分SCs和IPSCs/ESCs的凝集素，并在细胞糖蛋白水平上获得了多能性状态的特征。相关糖基转移酶基因的表达谱与凝集素芯片的结果一致。在38种凝集素中，rBC2LCN仅检测到未分化的IPSCs/ESCs，而未检测到分化的SCs。因此，高密度凝集素微阵列不仅可用于糖链的综合分析，还可用于细胞糖基概念下的干细胞诊断。
Induced pluripotent stem cells (iPSCs) can now be produced from various somatic cell (SC) lines by ectopic expression of the four transcription factors. Although the procedure has been demonstrated to induce global change in gene and microRNA expressions and even epigenetic modification, it remains largely unknown how this transcription factor-induced reprogramming affects the total glycan repertoire expressed on the cells. Here we performed a comprehensive glycan analysis using 114 types of human iPSCs generated from five different SCs and compared their glycomes with those of human embryonic stem cells (ESCs; nine cell types) using a high density lectin microarray. In unsupervised cluster analysis of the results obtained by lectin microarray, both undifferentiated iPSCs and ESCs were clustered as one large group. However, they were clearly separated from the group of differentiated SCs, whereas all of the four SCs had apparently distinct glycome profiles from one another, demonstrating that SCs with originally distinct glycan profiles have acquired those similar to ESCs upon induction of pluripotency. Thirty-eight lectins discriminating between SCs and iPSCs/ESCs were statistically selected, and characteristic features of the pluripotent state were then obtained at the level of the cellular glycome. The expression profiles of relevant glycosyltransferase genes agreed well with the results obtained by lectin microarray. Amongthe 38 lectins, rBC2LCN was found to detect only undifferentiated iPSCs/ESCs and not differentiated SCs. Hence, the high density lectin microarray has proved to be valid for not only comprehensive analysis of glycans but also diagnosis of stem cells under the concept of the cellular glycome.