Downstream targets of HOXB4 in a cell line model of primitive hematopoietic progenitor cells
Downstream targets of HOXB4 in a cell line model of primitive hematopoietic progenitor cells
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DOI：
10.1182/blood-2009-11-253872
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发表时间：
2010-08-05
期刊：
影响因子：
20.3
通讯作者：
Forget, Bernard G.
中科院分区：
文献类型：
--
作者：
Lee, Han M.;Zhang, Hui;Forget, Bernard G.
同源框转录因子HOXB 4的增强表达已显示增强造血干细胞的离体和体内自我更新和扩增。为了研究HOXB 4在造血祖细胞中的下游靶点，在原始造血祖细胞系EML中组成型过表达HOXB 4。使用了两种全基因组分析技术：使用微阵列和染色质免疫沉淀（ChIP）芯片的RNA表达谱。RNA表达谱分析显示，与仅用载体转导的对照KLS-EML细胞相比，在过表达HOXB 4的KLS（c-Kit（+），Lin（-），Sca-1（+））-EML细胞（KLS-EML-HOXB 4）中差异表达465个基因转录物。特别是，观察到红系特异性基因转录物在KLS-EMLHOXB 4细胞中高度下调。ChIP芯片分析显示，在KLS-EML-HOXB 4细胞中，HOXB 4占据了1910个基因的启动子区，如CD 34、Sox 4和B220。ChIP芯片和RNA表达谱数据集的并排比较提供了相关信息，并将Gp 49 a和Laptm 4 b鉴定为候选的“干相关”基因。这两个基因在两个数据集列表中排名很高，并且先前已显示优先在造血干细胞中表达并在成熟造血细胞中下调，从而使它们成为造血细胞中未来功能研究的有吸引力的候选者。（血。2010;116（5）：720-730）
Enforced expression of the homeobox transcription factor HOXB4 has been shown to enhance hematopoietic stem cell self-renewal and expansion ex vivo and in vivo. To investigate the downstream targets of HOXB4 in hematopoietic progenitor cells, HOXB4 was constitutively overexpressed in the primitive hematopoietic progenitor cell line EML. Two genome-wide analytical techniques were used: RNA expression profiling using microarrays and chromatin immuno-precipitation (ChIP)-chip. RNA expression profiling revealed that 465 gene transcripts were differentially expressed in KLS (c-Kit(+), Lin(-), Sca-1(+))-EML cells that overexpressed HOXB4 (KLS-EML-HOXB4) compared with control KLS-EML cells that were transduced with vector alone. In particular, erythroid-specific gene transcripts were observed to be highly down-regulated in KLS-EMLHOXB4 cells. ChIP-chip analysis revealed that the promoter region for 1910 genes, such as CD34, Sox4, and B220, were occupied by HOXB4 in KLS-EML-HOXB4 cells. Side-by-side comparison of the ChIP-chip and RNA expression profiling datasets provided correlative information and identified Gp49a and Laptm4b as candidate "stemness-related" genes. Both genes were highly ranked in both dataset lists and have been previously shown to be preferentially expressed in hematopoietic stem cells and down-regulated in mature hematopoietic cells, thus making them attractive candidates for future functional studies in hematopoietic cells. (Blood. 2010;116(5):720-730)