Lentiviral gene transfer regenerates hematopoietic stem cells in a mouse model for Mpl-deficient aplastic anemia
Lentiviral gene transfer regenerates hematopoietic stem cells in a mouse model for Mpl-deficient aplastic anemia
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DOI：
10.1182/blood-2010-09-308262
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发表时间：
2011-04-07
期刊：
影响因子：
20.3
通讯作者：
Modlich, Ute
中科院分区：
文献类型：
--
作者：
Heckl, Dirk;Wicke, Daniel C.;Modlich, Ute
Thpo/Mpl信号除了在巨核生成中发挥作用外，在造血干细胞（hsc）的维持中也起着重要作用。Mpl失活突变的患者由于造血干细胞的进行性丢失而发生血小板减少症和再生障碍性贫血。然而，目前尚不清楚造血干细胞的损失是否是一个不可逆的过程。在这项研究中，我们使用Mpl敲除（Mpl(-/-)）小鼠模型，并从新开发的慢病毒载体特异性地在Mpl生理靶群，即造血干细胞和巨核细胞中表达Mpl。在使用慢病毒eGFP报告载体验证了在体内的谱系特异性表达后，我们将转导的Mpl（-/-）骨髓细胞移植到Mpl（-/-）小鼠中。我们发现，从转录靶向载体中恢复Mpl表达可以防止Mpl异位表达的致命不良反应，补充HSC库，恢复干细胞特性，并纠正血小板产生。在一些小鼠中，巨核细胞计数异常高，伴有骨新生和骨髓纤维化。基因校正的Mpl（-/-）细胞增加了长期的再填充潜力，重组小鼠的谱系（-）Sca1(+)cKit（+）细胞和早期祖细胞群体显著增加。对mpl校正小鼠谱系（-）Sca1(+)cKit（+）细胞的转录组分析显示，参与HSC自我更新的基因发生了功能调整。(血。2011;117 (14):3737 - 3747)
Thpo/Mpl signaling plays an important role in the maintenance of hematopoietic stem cells (HSCs) in addition to its role in megakaryopoiesis. Patients with inactivating mutations in Mpl develop thrombocytopenia and aplastic anemia because of progressive loss of HSCs. Yet, it is unknown whether this loss of HSCs is an irreversible process. In this study, we used the Mpl knockout (Mpl(-/-)) mouse model and expressed Mpl from newly developed lentiviral vectors specifically in the physiologic Mpl target populations, namely, HSCs and megakaryocytes. After validating lineage-specific expression in vivo using lentiviral eGFP reporter vectors, we performed bone marrow transplantation of transduced Mpl(-/-) bone marrow cells into Mpl(-/-) mice. We show that restoration of Mpl expression from transcriptionally targeted vectors prevents lethal adverse reactions of ectopic Mpl expression, replenishes the HSC pool, restores stem cell properties, and corrects platelet production. In some mice, megakaryocyte counts were atypically high, accompanied by bone neo-formation and marrow fibrosis. Gene-corrected Mpl(-/-) cells had increased long-term re-populating potential, with a marked increase in lineage(-)Sca1(+)cKit(+) cells and early progenitor populations in reconstituted mice. Transcriptome analysis of lineage(-)Sca1(+)cKit(+) cells in Mpl-corrected mice showed functional adjustment of genes involved in HSC self-renewal. (Blood. 2011;117(14):3737-3747)