Verhoeyen Els, Wiznerowicz Maciej, Olivier Delphine, Izac Brigitte, Trono Didier, Dubart-Kupperschmitt Anne, Cosset François-Loïc
INSERM, U412, Lyon, France.
Blood. 2005 Nov 15;106(10):3386-95. doi: 10.1182/blood-2004-12-4736. Epub 2005 Aug 2.
A major limitation of current lentiviral vectors (LVs) is their inability to govern efficient gene transfer into quiescent cells, such as human CD34(+) cells, that reside in the G(0) phase of the cell cycle and that are highly enriched in hematopoietic stem cells. This hampers their application for gene therapy of hematopoietic cells. Here, we designed novel LVs that overcome this restriction by displaying "early-acting cytokines" on their surface. Display of thrombopoietin, stem cell factor, or both cytokines on the LV surface allowed efficient gene delivery into quiescent cord blood CD34(+) cells. Moreover, these surface-engineered LVs preferentially transduced and promoted survival of resting CD34(+) cells rather than cycling cells. Finally, and most importantly, these novel LVs allowed superior gene transfer in the most immature CD34(+) cells as compared to conventional LVs, even when the latter vectors were used to transduce cells in the presence of recombinant cytokines. This was demonstrated by their capacity to promote selective transduction of CD34(+) cell in in vitro derived long-term culture-initiating cell (LTC-IC) colonies and of long-term NOD/SCID repopulating cells (SRCs) in vivo.
当前慢病毒载体（LVs）的一个主要局限性在于它们无法有效地将基因导入静止细胞，如人类CD34(+)细胞，这些细胞处于细胞周期的G(0)期，并且在造血干细胞中高度富集。这限制了它们在造血细胞基因治疗中的应用。在此，我们设计了新型慢病毒载体，通过在其表面展示“早期作用细胞因子”来克服这一限制。在慢病毒载体表面展示血小板生成素、干细胞因子或这两种细胞因子，能够有效地将基因传递到静止的脐带血CD34(+)细胞中。此外，这些经过表面工程改造的慢病毒载体优先转导并促进静止CD34(+)细胞而非增殖细胞的存活。最后，也是最重要的一点，与传统慢病毒载体相比，这些新型慢病毒载体能够在最不成熟的CD34(+)细胞中实现更高效的基因传递，即使在使用重组细胞因子转导细胞时使用传统慢病毒载体也是如此。这一点通过它们在体外衍生的长期培养起始细胞（LTC-IC）集落中促进CD34(+)细胞的选择性转导以及在体内促进长期NOD/SCID重建造血细胞（SRCs）的能力得到了证明。
