In vivo gene transfer into adult stem cells in unconditioned mice by in situ delivery of a lentiviral vector.
In vivo gene transfer into adult stem cells in unconditioned mice by in situ delivery of a lentiviral vector.
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通过慢病毒载体的原位递送将基因体内基因转移到未条件化小鼠的成体干细胞中。
DOI：
10.1016/j.ymthe.2006.05.014
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Pan,Dao
中科院分区：
文献类型：
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作者：
Worsham,DNicole;Schuesler,Todd;vonKalle,Christof;Pan,Dao
体内病毒介导的骨髓干细胞基因转移通过骨腔注射的潜力，可以充分利用任何来源的干细胞存在，以前没有探索。这种方法可以避免体内造血干细胞（HSC）基因转移所遇到的一些困难。我们试图确定是否可以实现有效的基因转移，在HSC和间充质干/祖细胞（MSC）的慢病毒载体在小鼠股内注射。注射后4个月，在骨髓和淋巴亚群中观察到高达12%的GFP表达细胞。在Lin-c-kit + Sca 1 +HSC/祖细胞和具有多谱系潜能的CFU中观察到显著的转导效率，这也通过祖细胞衍生集落的双重PCR分析得到证实。二次BMT后4个月，我们在所有受体中观察到8.1%至15%的载体+CFU。LAM-PCR整合分析表明，多个转导克隆有助于这些动物的造血。我们还发现，表达GFP的MSC保留了多向分化潜能，在注射和BMT受体中检测到2.9%至8.8%的含GFP的CFU成纤维细胞。我们的数据提供的证据表明，在骨髓中的成体干细胞可以有效地转导“原位”体内载体管理，而无需预处理。这种方法可能导致治疗人类疾病的新应用。
The potential ofin vivolentivirus-mediated bone marrow stem cell gene transfer by bone cavity injection, which could take full advantage of any source of stem cells present there, has not been previously explored. Such an approach may avoid several difficulties encountered byex vivohematopoietic stem cell (HSC) gene transfer. We sought to determine if efficient gene transfer could be achieved in HSC and mesenchymal stem/progenitor cells (MSC) by intrafemoral injection of a lentivirus vector in mice. Four months after injection, up to 12% GFP-expressing cells were observed in myeloid and lymphoid subpopulations. Significant transduction efficiencies were seen in Lin−c-kit+Sca1+HSC/progenitors and CFU with multilineage potential, which were also confirmed by duplex PCR analysis of progenitor-derived colonies. Four months after secondary BMT, we observed 8.1 to 15% vector+CFU in all recipients. Integration analysis by LAM-PCR demonstrated that multiple transduced clones contributed to hematopoiesis in these animals. We also showed that GFP-expressing MSC retained multilineage differentiation potential, with 2.9 to 8.8% GFP-containing CFU-fibroblasts detected in both injected and BMT recipients. Our data provide evidence that adult stem cells in bone marrow can be efficiently transduced "in situ" byin vivovector administration without preconditioning. This approach could lead to a novel application for treatment of human diseases.