A Loss of Function Screen of Identified Genome-Wide Association Study Loci Reveals New Genes Controlling Hematopoiesis
A Loss of Function Screen of Identified Genome-Wide Association Study Loci Reveals New Genes Controlling Hematopoiesis
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DOI：
10.1371/journal.pgen.1004450
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发表时间：
2014-07-01
期刊：
影响因子：
4.5
通讯作者：
Cvejic, Ana
中科院分区：
文献类型：
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作者：
Bielczyk-Maczynska, Ewa;Serbanovic-Canic, Jovana;Cvejic, Ana
造血干细胞形成成熟细胞的过程在细胞水平上已经得到了很好的理解，我们也知道许多控制命运决定的关键转录因子。然而，许多上游信号传导和下游效应过程仅部分了解。全基因组关联研究（GWAS）在为剖析这些途径提供新的方向方面特别有用。一项GWAS荟萃分析确定了68个控制血小板大小和数量的遗传位点。然而，这些基因中只有四分之一是已知的造血调节因子。为了确定其余基因的功能，我们在斑马鱼中使用反义吗啉代寡核苷酸（MO）敲低蛋白质表达进行了中等通量遗传筛选，然后使用广泛的不同造血标记物对选定的基因进行组织学分析。由初始敲低产生的信息被用于分析表型并在造血中分层定位候选基因。对brd3a的进一步分析揭示了其在血小板分化中的重要作用，而不是维持和存活。使用从GWAS到功能的策略，我们不仅确定了一系列代表血小板生成和造血的新调节因子的基因，而且据我们所知，这项工作也代表了功能性遗传筛选策略的第一个例子，这是从GWA研究中获得生物学相关功能数据的关键一步。
The formation of mature cells by blood stem cells is very well understood at the cellular level and we know many of the key transcription factors that control fate decisions. However, many upstream signalling and downstream effector processes are only partially understood. Genome wide association studies (GWAS) have been particularly useful in providing new directions to dissect these pathways. A GWAS meta-analysis identified 68 genetic loci controlling platelet size and number. Only a quarter of those genes, however, are known regulators of hematopoiesis. To determine function of the remaining genes we performed a medium-throughput genetic screen in zebrafish using antisense morpholino oligonucleotides (MOs) to knock down protein expression, followed by histological analysis of selected genes using a wide panel of different hematopoietic markers. The information generated by the initial knockdown was used to profile phenotypes and to position candidate genes hierarchically in hematopoiesis. Further analysis of brd3a revealed its essential role in differentiation but not maintenance and survival of thrombocytes. Using the from-GWAS-to-function strategy we have not only identified a series of genes that represent novel regulators of thrombopoiesis and hematopoiesis, but this work also represents, to our knowledge, the first example of a functional genetic screening strategy that is a critical step toward obtaining biologically relevant functional data from GWA study for blood cell traits.