Alonso-Ferrero Maria E, van Til Niek P, Bartolovic Kerol, Mata Márcia F, Wagemaker Gerard, Moulding Dale, Williams David A, Kinnon Christine, Waddington Simon N, Milsom Michael D, Howe Steven J
Molecular and Cellular Immunology, University College London Great Ormond Street Institute of Child Health, London, UK.
Department of Hematology, Erasmus University Medical Center, Rotterdam, The Netherlands; Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, The Netherlands.
Exp Hematol. 2018 Jan;57:21-29. doi: 10.1016/j.exphem.2017.09.003. Epub 2017 Sep 11.
Integration-deficient lentiviruses (IdLVs) deliver genes effectively to tissues but are lost rapidly from dividing cells. This property can be harnessed to express transgenes transiently to manipulate cell biology. Here, we demonstrate the utility of short-term gene expression to improve functional potency of hematopoietic stem and progenitor cells (HSPCs) during transplantation by delivering HOXB4 and Angptl3 using IdLVs to enhance the engraftment of HSPCs. Constitutive overexpression of either of these genes is likely to be undesirable, but the transient nature of IdLVs reduces this risk and those associated with unsolicited gene expression in daughter cells. Transient expression led to increased multilineage hematopoietic engraftment in in vivo competitive repopulation assays without the side effects reported in constitutive overexpression models. Adult stem cell fate has not been programmed previously using IdLVs, but we demonstrate that these transient gene expression tools can produce clinically relevant alterations or be applied to investigate basic biology.
整合缺陷型慢病毒（IdLVs）能有效地将基因传递到组织中，但在分裂细胞中会迅速丢失。这一特性可用于瞬时表达转基因以操纵细胞生物学。在此，我们通过使用IdLVs递送HOXB4和血管生成素样蛋白3（Angptl3）来增强造血干细胞和祖细胞（HSPCs）的植入，从而证明了短期基因表达在改善HSPCs移植过程中的功能效力方面的实用性。持续过表达这两种基因中的任何一种可能都不理想，但IdLVs的瞬时特性降低了这种风险以及与子代细胞中意外基因表达相关的风险。在体内竞争性再增殖试验中，瞬时表达导致多谱系造血植入增加，且无持续过表达模型中报道的副作用。此前尚未使用IdLVs对成体干细胞命运进行编程，但我们证明这些瞬时基因表达工具可产生临床相关的改变或用于研究基础生物学。
