Ohashi T, Boggs S, Robbins P, Bahnson A, Patrene K, Wei F S, Wei J F, Li J, Lucht L, Fei Y
Department of Human Genetics, University of Pittsburgh, PA 15261.
Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11332-6. doi: 10.1073/pnas.89.23.11332.
A recombinant retroviral vector (MFG-GC) was used to study the efficiency of transduction of the human gene encoding glucocerebrosidase (GC; D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45), in mouse hematopoietic stem cells and expression in their progeny. Transfer of the GC gene to CFU-S (spleen cell colony-forming units) in primary and secondary recipients was virtually 100%. In mice 4-7 months after transplantation, highly efficient transfer of the human gene to bone marrow cells capable of long-term reconstitution was confirmed by detection of one or two copies per mouse genome in hematopoietic tissues and in cultures of pure macrophages. Expression of the human gene exceeded endogenous activity by several fold in primary and secondary CFU-S, tissues from long-term reconstituted mice, and explanted macrophages cultures. These studies are evidence of the feasibility of efficient transfer of the GC gene to hematopoietic stem cells and expression in their progeny for many months after reconstitution. The results of this study strengthen the rationale for gene therapy as a treatment for Gaucher disease.
一种重组逆转录病毒载体（MFG-GC）被用于研究编码葡萄糖脑苷脂酶（GC；D-葡萄糖基-N-酰基鞘氨醇葡萄糖水解酶，EC 3.2.1.45）的人类基因在小鼠造血干细胞中的转导效率及其在子代中的表达。将GC基因转移至初代和二代受体的脾集落形成单位（CFU-S）中的效率几乎达到100%。在移植后4至7个月的小鼠中，通过在造血组织和纯巨噬细胞培养物中检测到每只小鼠基因组中有一到两个拷贝，证实了人类基因高效转移至能够长期重建的骨髓细胞中。在初代和二代CFU-S、长期重建小鼠的组织以及分离培养的巨噬细胞中，人类基因的表达比内源性活性高出数倍。这些研究证明了将GC基因高效转移至造血干细胞并在重建后数月内在其子代中表达的可行性。本研究结果强化了基因治疗作为戈谢病治疗方法的理论依据。
