Deficiency of ribosomal protein S19 in CD34+ cells generated by siRNA blocks erythroid development and mimics defects seen in Diamond-Blackfan anemia
Deficiency of ribosomal protein S19 in CD34+ cells generated by siRNA blocks erythroid development and mimics defects seen in Diamond-Blackfan anemia
复制标题
DOI：
10.1182/blood-2004-08-3115
复制
发表时间：
2005-06-15
期刊：
影响因子：
20.3
通讯作者：
Karlsson, S
中科院分区：
文献类型：
--
作者：
Flygare, J;Kiefer, T;Karlsson, S
Diamond-Blackfan 贫血 (DBA) 是一种先天性红细胞再生障碍性贫血，其中 25% 的患者存在核糖体蛋白 S19 (RPS19) 基因突变。为了研究 RPS19 缺陷对造血的影响，我们用 3 个表达针对 RPS19 的小干扰 RNA (siRNA) 的慢病毒载体和 1 个乱序对照载体转导 CD34(+) 脐带血 (CB) 和骨髓 (BM) 细胞。所有载体还表达绿色荧光蛋白（GFP）。 siRNA载体转导不同程度地降低了RPS19 mRNA水平，导致红细胞缺陷，与RPS19下调程度相关，并通过表达siRNA抗性RPS19转录物来挽救。 CD34(+)CB和BM细胞中红系集落形成能力与RPS19水平共同降低。在支持红系分化的液体培养中，RPS19沉默的以及DBA患者CD34(+)细胞表现出增殖能力降低和红系分化受损，导致红系集落形成单位(CFU-E)减少。在检测骨髓发育时，发现对增殖的影响不太明显。这项研究首次表明RPS19沉默会降低造血祖细胞的增殖能力并导致红系发育缺陷。 (c) 2005 年，美国血液学会。
Diamond-Blackfan anemia (DBA) is a congenital red cell aplasia in which 25% of the patients have a mutation in the ribosomal protein S19 (RPS19) gene. To study effects of RPS19 deficiency in hematopoiesis we transduced CD34(+) umbilical cord blood (CB) and bone marrow (BM) cells with 3 lentiviral vectors expressing small interfering RNA (siRNA) against RPS19 and 1 scrambled control vector. All vectors also express green fluorescent protein (GFP). Transduction with the siRNA vectors reduced RPS19 mRNA levels to various degrees, which resulted in erythroid defects, correlating to the degree of RPS19 down-regulation, and was rescued by expression of an siRNA-resistant RPS19 transcript. Erythroid colony formation capacity conjointly decreased with RPS19 levels in CD34(+) CB and BM cells. In liquid culture supporting erythroid differentiation, RPS19-slienced as well as DBA patient CD34(+) cells exhibited reduced proliferative capacity and impaired erythroid differentiation resulting in fewer erythroid colony-forming units (CFU-Es). When assaying myeloid development, a less pronounced influence on proliferation was seen. This study shows for the first time that RPS19 silencing decreases the proliferative capacity of hematopoietic progenitors and leads to a defect in erythroid development. (c) 2005 by The American Society of Hematology.