Proliferation deficiency of multipotent hematopoietic progenitors in ribosomal protein S19 (RPS19)-deficient Diamond-Blackfan anemia improves following RPS19 gene transfer
Proliferation deficiency of multipotent hematopoietic progenitors in ribosomal protein S19 (RPS19)-deficient Diamond-Blackfan anemia improves following RPS19 gene transfer
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DOI：
10.1016/s1525-0016(03)00091-1
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发表时间：
2003-05-01
影响因子：
12.4
通讯作者：
Karlsson, S
中科院分区：
文献类型：
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作者：
Hamaguchi, I;Flygare, J;Karlsson, S
Diamond-Blackfan贫血（DBA）是一种先天性骨髓衰竭综合征，其特征是红系祖细胞的特异性缺乏。由于一些DBA患者随着年龄的增长血小板和粒细胞减少，我们询问DBA患者的多能造血祖细胞在液体扩增培养中是否具有正常的增殖能力。DBA患者的CD 34（+）细胞在含有IL-3、IL-6和SCF的液体培养中表现出增殖缺陷。在含有IL-3、IL-6、SCF、Tpo、FIL和G-CSF或含有IL-3、IL-6和SCF的有限稀释试验中，DBA患者的单个CD 34（+）CD 38（-）细胞表现出增殖募集不足。我们的研究结果表明，潜在的造血缺陷DBA可能不仅限于红系。由于一部分DBA患者存在核糖体蛋白S19（RPS 19）缺陷，我们构建了含有RPS 19基因的慢病毒载体，用于在RPS 19缺陷型DBA患者的造血祖细胞中过表达。RPS 19转基因的增强表达改善了来自具有RPS 19突变的DBA患者的CD 34（+）细胞的增殖。类似地，如通过集落测定和红系分化培养所确定的，RPS 19的强制表达改善了RPS 19缺陷型造血祖细胞的红系发育。这些结果表明，基因治疗RPS 19缺陷型DBA是可行的。
Diamond-Blackfan anemia (DBA) is a congenital bone marrow failure syndrome characterized by a specific deficiency in erythroid progenitors. Since some patients with DBA develop a reduction in thrombocytes and granulocytes with age, we asked whether multipotent hematopoietic progenitors from DBA patients had normal proliferative capacity in liquid expansion cultures. CD34(+) cells derived from DBA patients showed deficient proliferation in liquid culture containing IL-3, IL-6, and SCF. Single CD34(+) CD38(-) cells from DBA patients exhibited deficient proliferation recruitment in a limiting dilution assay containing IL-3, IL-6, SCF, Tpo, FIL, and G-CSF or containing IL-3, IL-6, and SCF. Our findings suggest that the underlying hematopoietic defect in DBA may not be limited to the erythroid lineage. Since a fraction of DBA patients have a deficiency in ribosomal protein S19 (RPS19), we constructed lentiviral vectors containing the RPS19 gene for overexpression in hematopoietic progenitors from RPS19-deficient DBA patients. Enforced expression of the RPS19 transgene improved the proliferation of CD34(+) cells from DBA patients with RPS19 mutation. Similarly, enforced expression of RPS19 improved erythroid development of RPS19-deficient hematopoietic progenitors as determined by colony assays and erythroid differentiation cultures. These findings suggest that gene therapy for RPS19-deficient DBA is feasible.