Histone deacetylase inhibitor valproic acid enhances the cytokine-induced expansion of human hematopoietic stem cells
Histone deacetylase inhibitor valproic acid enhances the cytokine-induced expansion of human hematopoietic stem cells
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DOI：
10.1158/0008-5472.can-04-3063
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发表时间：
2005-02-15
期刊：
影响因子：
11.2
通讯作者：
Nervi, C
中科院分区：
文献类型：
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作者：
De Felice, LD;Tatarelli, C;Nervi, C
在不丧失自我更新能力的前提下体外扩增人造血干细胞(HSC)是移植、基因和细胞治疗的重要靶点。丙戊酸是一种安全且广泛使用的神经系统药物，可有效抑制历史上的脱乙酰酶活性。在这里，我们发现丙戊酸加入从脐带血、动员的外周血和骨髓中分离的人CD34+细胞的液体培养中，从形态、细胞化学和免疫表型上显示出强烈地增强了不同细胞因子组合的体外扩增潜力。克隆形成和基因表达分析。值得注意的是，丙戊酸在两种(Flt3L+血小板生成素)或四种细胞因子(Flt3L+血小板生成素+干细胞因子+白介素3)的扩增培养1周(范围40-89%)或3周(范围21-52%)扩增培养后高度保持CD34阳性。此外，丙戊酸处理增加了HOXB4和AC133上特定调控位点的组蛋白H4乙酰化水平。HOXB4是一种转录因子基因，在HSC自我更新调节中起关键作用，AC133是公认的干细胞群体标记基因。总体而言，我们的结果将丙戊酸引起的染色质可及性的变化与细胞因子在维持和扩大原始造血干细胞种群方面的作用增强联系在一起。这些发现强调了新的表观遗传学方法在体外改变HSC命运的可能性。
Ex vivo amplification of human hematopoietic stem cells (HSC) without loss of their self-renewing potential represents art important target for transplantation, gene and cellular therapies. Valproic acid is a safe and widely used neurologic agent that acts as a potent inhibitor of historic deacetylase activities. Here, we show that valproic acid addition to liquid cultures of human CD34+ cells isolated from cord blood, mobilized peripheral blood, and bone marrow strongly enhances the ex vivo expansion potential of different cytokine cocktails as shown by morphologic, cytochemical, immunophenotypical. clonogenic, and gene expression analyses. Notably, valproic acid highly preserves the CD34 positivity after 1 week (range, 40-89%) or 3 weeks (range, 21-52%) amplification cultures with two (Flt3L + thrombopoietin) or four cytokines (Flt3L + thrombopoietin + stem cell factor + interleukin 3). Moreover, valproic acid treatment increases histone H4 acetylation levels at specific regulatory sites on HOXB4, a transcription factor gene with a key role in the regulation of HSC self-renewal and AC133, a recognized marker gene for stem cell populations. Overall, our results relate the changes induced by valproic acid on chromatin accessibility with the enhancement of the cytokine effect on the maintenance and expansion of a primitive hematopoietic stem cell population. These findings underscore the potentiality of novel epigenetic approaches to modify HSC fate in vitro.