The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers
The Lgr5 intestinal stem cell signature: robust expression of proposed quiescent '+4' cell markers
复制标题
DOI：
10.1038/emboj.2012.166
复制
发表时间：
2012-07-18
期刊：
影响因子：
11.4
通讯作者：
Clevers, Hans
中科院分区：
文献类型：
--
作者：
Munoz, Javier;Stange, Daniel E.;Clevers, Hans
目前在小肠隐窝中定义了两种类型的干细胞：循环隐窝基柱状(CBC)细胞和静止的‘+4’细胞。在这里，我们将转录组学和蛋白质组学结合起来，为Lgr5(+)CBC细胞定义一个明确的分子特征。使用两个微阵列平台对FACS分选的Lgr5(+)干细胞及其子代进行转录图谱分析，揭示了384个独特基因的mRNA干细胞特征。对同一细胞群体的定量质谱分析发现，肠道干细胞中富含278种蛋白质。信使核糖核酸和蛋白质数据集显示了高度的相关性，并定义了510个干细胞丰富基因的组合签名。通过mRNA原位杂交进一步研究了Lgr5(+)干细胞的空间表达模式，发现大约一半的基因呈梯度表达，在隐窝底部表达水平最高，另一半在Lgr5(+)干细胞中唯一表达。利用新建立的敲入小鼠对其中一个标志性基因Smoc2进行血统追踪，证实了其干细胞特异性。利用这个资源，我们发现-并通过独立的方法证实-建议的静止/‘+4’干细胞标记Bmi1，TERT，Hopx和Lrig1在CBC细胞中强健表达。EMBO期刊(2012)31,3079-3091。DOI：10.1038/Intemj.2012.166；2012年6月12日在线发布
Two types of stem cells are currently defined in small intestinal crypts: cycling crypt base columnar (CBC) cells and quiescent '+4' cells. Here, we combine transcriptomics with proteomics to define a definitive molecular signature for Lgr5(+) CBC cells. Transcriptional profiling of FACS-sorted Lgr5(+) stem cells and their daughters using two microarray platforms revealed an mRNA stem cell signature of 384 unique genes. Quantitative mass spectrometry on the same cell populations identified 278 proteins enriched in intestinal stem cells. The mRNA and protein data sets showed a high level of correlation and a combined signature of 510 stem cell-enriched genes was defined. Spatial expression patterns were further characterized by mRNA in-situ hybridization, revealing that approximately half of the genes were expressed in a gradient with highest levels at the crypt bottom, while the other half was expressed uniquely in Lgr5(+) stem cells. Lineage tracing using a newly established knock-in mouse for one of the signature genes, Smoc2, confirmed its stem cell specificity. Using this resource, we find-and confirm by independent approaches-that the proposed quiescent/'+4' stem cell markers Bmi1, Tert, Hopx and Lrig1 are robustly expressed in CBC cells. The EMBO Journal (2012) 31, 3079-3091. doi:10.1038/emboj.2012.166; Published online 12 June 2012