Effect of increased HoxB4 on human megakaryocytic development
Effect of increased HoxB4 on human megakaryocytic development
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DOI：
10.1016/j.bbrc.2010.06.075
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发表时间：
2010-07-30
影响因子：
3.1
通讯作者：
Lasky, Larry C.
中科院分区：
文献类型：
--
作者：
Zhong, Yiming;Sullenbarger, Brent;Lasky, Larry C.
为了离体产生临床有用量的血小板，我们可能需要首先增强造血干细胞（HSC）和/或巨核细胞（Mk）祖细胞的早期自我更新。同源域转录因子HoxB 4已被证明是干细胞更新和造血的重要调节因子;然而，其对巨核细胞生成的影响尚不清楚。在这项研究中，我们研究了HoxB 4过表达或RNA沉默对人TF 1祖细胞系中巨核细胞发育的影响;然后，我们使用重组tPTD-HoxB 4融合蛋白来研究外源性HoxB 4对人CD 34阳性选择的脐带血细胞的巨核细胞发育的影响。我们发现TF 1细胞中异位HoxB 4增加了CD 61和CD 41 a的抗原表达，增加了血小板生成素受体（TpoR）、Scl-1、细胞周期蛋白D1、Fog-1和Fli-1的基因表达，同时降低了c-Myb的表达。HoxB 4 RNA沉默可降低TF 1细胞中CD 61和CD 41 a的表达，降低Fli-1的表达，而增加c-Myb的表达。重组tPTD-HoxB 4融合蛋白可增加CD 34阳性脐血细胞向巨核细胞分化过程中CD 41 a和CD 61阳性细胞的百分率和绝对数，增加巨核细胞集落形成单位（CFU-Mk）的数量。加入tPTD-HoxB 4融合蛋白可增加TpoR、Cyclin D1、Fog-1和Fli-1的基因表达，而抑制c-Myb的表达。我们的数据表明，增加HoxB 4增强人TF 1细胞和CD 34阳性选择的脐带血细胞的早期巨核细胞的发展，主要是通过上调TpoR和Fli-1的表达和下调c-Myb的表达。增加HoxB 4的表达或添加重组HoxB 4蛋白可能是扩增Mks以生产用于输血医学的血小板的一种方法。(C)2010年爱思唯尔公司All rights reserved.
In order to produce clinically useful quantities of platelets ex vivo we may need to firstly enhance early self-renewal of hematopoietic stem cells (HSCs) and/or megakaryocyte (Mk) progenitors. The homeodomain transcription factor HoxB4 has been shown to be an important regulator of stem cell renewal and hematopoiesis; however, its effect on megakaryopoiesis is unclear. In this study, we investigated the effect of HoxB4 overexpression or RNA silencing on megakaryocytic development in the human TF1 progenitor cell line; we then used recombinant tPTD-HoxB4 fusion protein to study the effect of exogenous HoxB4 on megakaryocytic development of human CD34 positively-selected cord blood cells. We found that ectopic HoxB4 in TF1 cells increased the antigen expression of CD61 and CD41a, increased the gene expression of thrombopoietin receptor (TpoR), Scl-1, Cyclin D1, Fog-1 and Fli-1 while it decreased c-Myb expression. HoxB4 RNA silencing in TF1 cells decreased the expression of CD61 and CD41a and decreased Fli-1 expression while it increased the expression of c-Myb. Recombinant tPTD-HoxB4 fusion protein increased the percentages and absolute numbers of CD41a and CD61 positive cells during megakaryocytic differentiation of CD34 positively-selected cord blood cells and increased the numbers of colony-forming unit-megakaryocyte (CFU-Mk). Adding tPTD-HoxB4 fusion protein increased the gene expression of TpoR, Cyclin D1, Fog-1 and Fli-1 while it inhibited c-Myb expression. Our data suggest that increased HoxB4 enhanced early megakaryocytic development in human TF1 cells and CD34 positively-selected cord blood cells primarily by upregulating TpoR and Fli-1 expression and downregulating c-Myb expression. Increasing HoxB4 expression or adding recombinant HoxB4 protein might be a way to expand Mks for the production of platelets for use in transfusion medicine. (C) 2010 Elsevier Inc. All rights reserved.