Comprehensive microRNA expression profiling of the hematopoietic hierarchy
Comprehensive microRNA expression profiling of the hematopoietic hierarchy
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DOI：
10.1073/pnas.1009320107
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发表时间：
2010-08-31
影响因子：
11.1
通讯作者：
Hansen, C. L.
中科院分区：
文献类型：
--
作者：
Petriv, O. I.;Kuchenbauer, F.;Hansen, C. L.
造血系统产生大量高度特化的细胞类型，这些细胞类型是通过分级分化过程从共同的干细胞群体衍生的。miRNAs是协调这种分化的关键参与者。在这里，我们报告了一种高通量微流控实时定量PCR（RT-qPCR）方法的开发和应用，该方法用于生成从正常成年小鼠造血组织中分离的27个表型不同的细胞群的全局miRNA谱。总共使用了80，000个RT-qPCR测定来绘制整个造血层次中miRNA表达的景观，包括罕见的祖细胞和干细胞群体。我们表明，miRNA的配置文件允许直接推断细胞谱系关系和功能相似性。我们的分析揭示了多能祖细胞和干细胞中miRNA表达模式的密切相关性，随后是限制分化潜能至单一谱系后的主要重编程。在单个造血细胞中的miRNA表达的分析进一步表明，miRNA表达在高度纯化的群体中受到非常严格的调控，强调了单细胞miRNA谱用于评估区室异质性的潜力。
The hematopoietic system produces a large number of highly specialized cell types that are derived through a hierarchical differentiation process from a common stem cell population. miRNAs are critical players in orchestrating this differentiation. Here, we report the development and application of a high-throughput microfluidic real-time quantitative PCR (RT-qPCR) approach for generating global miRNA profiles for 27 phenotypically distinct cell populations isolated from normal adult mouse hematopoietic tissues. A total of 80,000 RT-qPCR assays were used to map the landscape of miRNA expression across the hematopoietic hierarchy, including rare progenitor and stem cell populations. We show that miRNA profiles allow for the direct inference of cell lineage relations and functional similarity. Our analysis reveals a close relatedness of the miRNA expression patterns in multipotent progenitors and stem cells, followed by a major reprogramming upon restriction of differentiation potential to a single lineage. The analysis of miRNA expression in single hematopoietic cells further demonstrates that miRNA expression is very tightly regulated within highly purified populations, underscoring the potential of single-cell miRNA profiling for assessing compartment heterogeneity.