An RNA interference model of RPS 19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
An RNA interference model of RPS 19 deficiency in Diamond-Blackfan anemia recapitulates defective hematopoiesis and rescue by dexamethasone: identification of dexamethasone-responsive genes by microarray
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DOI：
10.1182/blood-2004-08-3313
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发表时间：
2005-06-15
期刊：
影响因子：
20.3
通讯作者：
Sieff, CA
中科院分区：
文献类型：
--
作者：
Ebert, BL;Lee, MM;Sieff, CA
Diamond-Blackfan贫血（DBA）是一种先天性红细胞减少症，是研究红细胞分化的典型疾病，但对其了解甚少。RPS19是唯一与DBA相关的基因，但其与红系分化的相关性尚不清楚。糖皮质激素刺激DBA患者红细胞生成的分子基础尚未确定。我们证明，通过逆转录病毒短发夹rna （shRNAs）的表达，RPS19转录物的靶向降解可以阻断培养的人CD34（+）细胞中红系祖细胞的增殖和分化。地塞米松治疗rps19缺陷细胞可使红细胞分化恢复到正常水平。我们研究了DBA药物治疗的分子基础，使用寡核苷酸微阵列来研究CD34（+）细胞在地塞米松、促红细胞生成素、干细胞因子和白细胞介素-3联合治疗下的基因表达。地塞米松没有改变RPS19的表达，但激活了包括一组关键造血调节基因的遗传程序。地塞米松可上调红系祖细胞特异性基因，而下调非红系谱系特异性基因。因此，缺乏RPS19会阻碍未成熟红系祖细胞的增殖，而地塞米松通过独立于RPS19的机制激活相同细胞群的增殖。(c) 2005年由美国血液学会出版。
Diamond-Blackfan anemia (DBA), a congenital erythroblastopenia, is a model disease for the study of erythroid differentiation but is poorly understood. RPS19 is the only gene yet to have been associated with DBA, but its relevance to erythroid differentiation is unclear. The molecular basis for the stimulation of erythropoiesis by glucocorticoids in patients with DBA has not been identified. We demonstrate that targeted degradation of the RPS19 transcript, through retroviral expression of short hairpin RNAs (shRNAs), blocks the proliferation and differentiation of erythroid progenitor cells in cultured human CD34(+) cells. Treatment of RPS19-deficient cells with dexamethasone restores erythroid differentiation to normal levels. We investigated the molecular basis of pharmacologic therapies for DBA using oligonucleotide microarrays to survey gene expression in CD34(+) cells treated with combinations of dexamethasone, erythropoietin, stem cell factor, and interleukin-3. Dexamethasone did not alter expression of RPS19 but activated a genetic program that includes a set of key hematopoietic regulatory genes. Genes specific to erythroid progenitor cells were up-regulated by dexamethasone, while genes specific to nonerythroid lineages were down-regulated. Deficiency of RPS19 therefore blocks proliferation of immature erythroid progenitor cells, and dexamethasone activates proliferation of the same cell population through mechanisms independent of RPS19. (c) 2005 by The American Society of Hematology.