Global transcriptome analyses of human and murine terminal erythroid differentiation
Global transcriptome analyses of human and murine terminal erythroid differentiation
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DOI：
10.1182/blood-2014-01-548305
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发表时间：
2014-05-29
期刊：
影响因子：
20.3
通讯作者：
Gallagher, Patrick G.
中科院分区：
文献类型：
--
作者：
An, Xiuli;Schulz, Vincent P.;Gallagher, Patrick G.
我们最近开发了基于荧光激活细胞分选（FACS）的方法来纯化形态学和功能离散的细胞群，每个细胞群代表终末红细胞分化的特定阶段。我们使用这些技术获得了处于不同发育阶段的人和鼠红细胞的纯群体。从这些细胞中制备RNA并进行RNA测序分析，创建无偏倚的阶段特异性转录组。来自不同阶段的转录组紧密聚类，甚至在生物学上不同的重复之间，验证了基于facs的分析的实用性。生物信息学分析显示，在分化阶段之间存在显著差异，在转录空间中定义每个阶段的基因表达谱既有相同的，也有不同的。在分化阶段，基因表达存在巨大的时间变化，每个阶段都表现出独特的转录组。聚类和网络分析表明，不同功能的基因在分化过程中观察到的不同阶段特异性表达模式丰富。人类和小鼠转录组之间存在许多差异，基因表达的整体模式存在显著差异。这些数据为正常和异常红细胞的研究提供了重要的资源，使我们能够更深入地了解各种遗传性和获得性红细胞疾病中红细胞的发育机制。
We recently developed fluorescence-activated cell sorting ( FACS)-based methods to purify morphologically and functionally discrete populations of cells, each representing specific stages of terminal erythroid differentiation. We used these techniques to obtain pure populations of both human and murine erythroblasts at distinct developmental stages. RNA was prepared from these cells and subjected to RNA sequencing analyses, creating unbiased, stage-specific transcriptomes. Tight clustering of transcriptomes from differing stages, even between biologically different replicates, validated the utility of the FACS-based assays. Bioinformatic analyses revealed that there were marked differences between differentiation stages, with both shared and dissimilar gene expression profiles defining each stage within transcriptional space. There were vast temporal changes in gene expression across the differentiation stages, with each stage exhibiting unique transcriptomes. Clustering and network analyses revealed that varying stage-specific patterns of expression observed across differentiation were enriched for genes of differing function. Numerous differences were present between human and murine transcriptomes, with significant variation in the global patterns of gene expression. These data provide a significant resource for studies of normal and perturbed erythropoiesis, allowing a deeper understanding of mechanisms of erythroid development in various inherited and acquired erythroid disorders.