Amsellem Sophie, Ravet Emmanuel, Fichelson Serge, Pflumio Françoise, Dubart-Kupperschmitt Anne
Institut Cochin, INSERM U567, CNRS UMR 8104, Hematology Department, Maternité de Port-Royal, Paris, France.
Mol Ther. 2002 Nov;6(5):673-7.
Gene therapy using permanent modifications of hematopoietic stem cells (HSC) has increasing potential applications for both genetic and acquired diseases. Considerable progress has been made recently in gene transfer to HSC by the use of lentivirus-derived vectors, which have the capacity to transduce noncycling cells. However, overall efficiency of HSC transduction reported so far is still not sufficient for numerous applications. We describe here an improved HSC transduction protocol, using the previously described lentiviral vector, that leads to more than 90% transduction of human CD34+ cells from cord blood, including NOD-LtSz-scid/scid repopulating cells. Moreover, under the same conditions, we transduce more than 75% and 80% of CD34+ cells mobilized in peripheral blood and from bone marrow, respectively. We further show that transgene expression is stable through time and hematopoietic cell differentiation in vitro as well as in vivo. Such a high HSC transduction efficiency opens new opportunities for both gene therapy applications and functional studies of regulator proteins of hematopoiesis.
对造血干细胞（HSC）进行永久性修饰的基因治疗在遗传疾病和后天性疾病方面的潜在应用日益增多。最近，通过使用慢病毒衍生载体将基因导入造血干细胞取得了相当大的进展，这些载体能够转导非循环细胞。然而，迄今为止报道的造血干细胞转导的总体效率对于众多应用来说仍然不够。我们在此描述一种改进的造血干细胞转导方案，使用先前描述的慢病毒载体，该方案可使来自脐带血的人类CD34+细胞的转导率超过90%，包括NOD-LtSz-scid/scid重建造血的细胞。此外，在相同条件下，我们分别使动员到外周血和骨髓中的CD34+细胞的转导率超过75%和80%。我们进一步表明，转基因表达在体外和体内随时间以及造血细胞分化都是稳定的。如此高的造血干细胞转导效率为基因治疗应用和造血调节蛋白的功能研究开辟了新的机会。
