Schwarz E J, Alexander G M, Prockop D J, Azizi S A
Center for Gene Therapy, MCP Hahnemann University, Philadelphia, PA 19102, USA.
Hum Gene Ther. 1999 Oct 10;10(15):2539-49. doi: 10.1089/10430349950016870.
Bone marrow stromal cells can be used as an alternative source of cells for neural transplantation and repair. Here, the efficacy of genetically modified marrow stromal cells was examined in a rat model of Parkinson disease. Rat marrow stromal cells (rMSCs) and human marrow stromal cells (hMSCs) were genetically engineered by transduction with retroviruses encoding tyrosine hydroxylase (TH) and GTP cyclohydrolase I, the enzyme necessary for production of the tetrahydrobiopterin cofactor for TH (BH4). Transduced cells synthesized 3,4-dihydroxyphenylalanine (L-DOPA) in vitro and maintained their multipotentiality after retroviral transduction. To examine the cells in vivo, transduced rMSCs were injected into the striatum of 6-hydroxydopamine-lesioned rats. L-DOPA and metabolites were detected by microdialysis in the denervated striatum of rats that received doubly transduced rMSCs. Also, there was a significant reduction in apomorphine-induced rotation when compared with controls. The cells engrafted and survived for at least 87 days. However, expression of the transgenes ceased at about 9 days, an observation consistent with reports from other laboratories in which similar retroviruses were used to express transgenes in the brain.
骨髓基质细胞可作为神经移植和修复的替代细胞来源。在此，我们在帕金森病大鼠模型中检测了基因改造的骨髓基质细胞的功效。通过用编码酪氨酸羟化酶（TH）和GTP环化水解酶I的逆转录病毒转导，对大鼠骨髓基质细胞（rMSCs）和人骨髓基质细胞（hMSCs）进行基因工程改造，GTP环化水解酶I是TH（BH4）的四氢生物蝶呤辅因子产生所必需的酶。转导后的细胞在体外合成3,4-二羟基苯丙氨酸（L-DOPA），并在逆转录病毒转导后保持其多能性。为了在体内检测这些细胞，将转导后的rMSCs注射到6-羟基多巴胺损伤大鼠的纹状体中。通过微透析在接受双重转导的rMSCs的大鼠失神经纹状体中检测L-DOPA和代谢产物。此外，与对照组相比，阿扑吗啡诱导的旋转显著减少。这些细胞植入并存活了至少87天。然而，转基因的表达在大约9天时停止，这一观察结果与其他实验室的报告一致，在那些报告中使用了类似的逆转录病毒在大脑中表达转基因。
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