Modulation of p210BCR-ABL activity in transduced primary human hematopoietic cells controls lineage programming
Modulation of p210BCR-ABL activity in transduced primary human hematopoietic cells controls lineage programming
复制标题
DOI：
10.1182/blood.v99.9.3197
复制
发表时间：
2002-05-01
期刊：
影响因子：
20.3
通讯作者：
Eaves, C
中科院分区：
文献类型：
--
作者：
Chalandon, Y;Jiang, XY;Eaves, C
用人类癌基因对原代造血细胞进行逆转录病毒转导，为研究控制这些细胞正常增殖和分化的分子机制提供了一种强有力的方法。在这里，我们表明，原始人CD34（+）脐带血细胞，包括多能以及粒细胞生成和红细胞限制性祖细胞，可以用MSCV-BCR-ABL-IRES-GFP逆转录病毒有效转导，导致其后代持续表达非常高水平的酪氨酸磷酸化p210（BCR-ABL）。有趣的是，即使存在支持绿色荧光蛋白（GFP）转导的对照细胞专门产生粒细胞的生长因子，BCR-ABL转导的祖细胞亚群也会产生大量不依赖促红细胞生成素的终末分化红系细胞，并减少粒细胞的数量。对半固体和液体培养物中单个转导细胞产生的单个克隆的分析表明，所有类型的转导 CD34+ 细胞都会高频率地引发这种 BCR-ABL 诱导的红系分化反应，而与它们先前明显的谱系定型状态无关。其他实验表明，当细胞在 BCR-ABL 特异性酪氨酸激酶抑制剂 STI-571 存在下转导和维持时，这种红细胞分化反应在很大程度上被阻止。这些发现表明，在原代人造血细胞中过表达 BCR-ABL 可以通过 BCR-ABL 激酶依赖性机制激活明显粒细胞生成受限的细胞中的红系分化程序，从而为阐明人造血细胞谱系命运决定和人白血病不忠的潜在机制提供了一种新的分子工具。 (C) 2002 年，美国血液学会。
Retroviral transduction of primary hematopoietic cells with human oncogenes provides a powerful approach to investigating the molecular mechanisms controlling the normal proliferation and differentiation of these cells. Here we show that primitive human CD34(+) cord blood cells, including multipotent as well as granulopoietic- and erythroid-restricted progenitors, can be efficiently transduced with a MSCV-BCR-ABL-IRES-GFP retrovirus, resulting in the sustained expression by their progeny of very high levels of tyrosine phosphorylated p210(BCR-ABL). Interestingly, even in the presence of growth factors that supported the exclusive production of granulopoietic cells from green fluorescent protein (GFP)-transduced control cells, BCR-ABL-transduced progenitor subpopulations generated large numbers of erythropoietin-independent terminally differentiating erythroid cells and reduced numbers of granulopoietic cells. Analyses of individual clones generated by single transduced cells in both semisolid and liquid cultures showed this BCR-ABL-induced erythroid differentiation response to be elicited at a high frequency from all types of transduced CD34(+) cells independent of their apparent prior lineage commitment status. Additional experiments showed that this erythroid differentiation response was largely prevented when the cells were transduced and maintained in the presence of the BCR-ABL-specific tyrosine kinase inhibitor, STI-571. These findings indicate that overexpression of BCR-ABL in primary human hematopoietic cells can activate an erythroid differentiation program in apparently granulopoietic-restricted cells through a BCR-ABL kinase-dependent mechanism, thus providing a new molecular tool for elucidating mechanisms underlying lineage fate determination in human hematopoietic cells and infidelity in human leukemia. (C) 2002 by The American Society of Hematology.