Orlic D, Girard L J, Anderson S M, Pyle L C, Yoder M C, Broxmeyer H E, Bodine D M
Hematopoiesis Section, Laboratory of Gene Transfer, National Human Genome Research Institute (NHGRI), National Institutes of Health (NIH), Bethesda, MD, USA.
Blood. 1998 May 1;91(9):3247-54.
One obstacle to retrovirus-mediated gene therapy for human hematopoietic disorders is the low efficiency of gene transfer into pluripotent hematopoietic stem cells (HSC). We have previously shown a direct correlation between retrovirus receptor mRNA levels in mouse HSC and the efficiency with which they are transduced. In the present study, we assayed retrovirus receptor mRNA levels in a variety of mouse and human HSC populations to identify HSC which may be more competent for retrovirus transduction. The highest levels of amphotropic retrovirus receptor (amphoR) mRNA were found in cryopreserved human cord blood HSC. The level of amphoR mRNA in Lin- CD34(+) CD38(-) cells isolated from frozen cord blood was 12-fold higher than the level in fresh cord blood Lin- CD34(+) CD38(-) cells. In mice, the level of amphoR mRNA in HSC from the bone marrow (BM) of mice treated with stem cell factor and granulocyte-colony stimulating factor was 2.8- to 7.8-fold higher than in HSC from the BM of untreated mice. These findings suggest that HSC from frozen cord blood and cytokine-mobilized BM may be superior targets for amphotropic retrovirus transduction compared with HSC from untreated adult BM.
逆转录病毒介导的基因疗法用于治疗人类造血系统疾病的一个障碍是基因导入多能造血干细胞（HSC）的效率较低。我们之前已经表明，小鼠造血干细胞中逆转录病毒受体mRNA水平与其被转导的效率之间存在直接关联。在本研究中，我们检测了多种小鼠和人类造血干细胞群体中的逆转录病毒受体mRNA水平，以鉴定可能更易于被逆转录病毒转导的造血干细胞。在冷冻保存的人类脐带血造血干细胞中发现了最高水平的嗜异性逆转录病毒受体（amphoR）mRNA。从冷冻脐带血中分离出的Lin-CD34(+)CD38(-)细胞中amphoR mRNA的水平比新鲜脐带血Lin-CD34(+)CD38(-)细胞中的水平高12倍。在小鼠中，用干细胞因子和粒细胞集落刺激因子处理的小鼠骨髓（BM）中的造血干细胞中amphoR mRNA的水平比未处理小鼠骨髓中的造血干细胞高2.8至7.8倍。这些发现表明，与未处理的成年骨髓中的造血干细胞相比，冷冻脐带血和细胞因子动员的骨髓中的造血干细胞可能是嗜异性逆转录病毒转导的更好靶标。
