GENETICALLY MODIFIED SKIN FIBROBLASTS PERSIST LONG AFTER TRANSPLANTATION BUT GRADUALLY INACTIVATE INTRODUCED GENES
GENETICALLY MODIFIED SKIN FIBROBLASTS PERSIST LONG AFTER TRANSPLANTATION BUT GRADUALLY INACTIVATE INTRODUCED GENES
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DOI：
10.1073/pnas.88.4.1330
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发表时间：
1991-02-01
影响因子：
11.1
通讯作者：
MILLER, AD
中科院分区：
文献类型：
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作者：
PALMER, TD;ROSMAN, GJ;MILLER, AD
基因工程成纤维细胞已成功地用于在动物中产生治疗性蛋白质，但尚未实现蛋白质的持续生产。 这限制了成纤维细胞介导的基因治疗在人类中的潜力。 我们已经研究了使用逆转录病毒载体携带编码人腺苷脱氨酶和新霉素磷酸转移酶的基因，在大鼠生产下降的现象。 虽然含有载体序列的移植皮肤成纤维细胞以恒定水平持续至少8.5个月，但1个月后载体表达降低> 1500倍。 在移植动物中未检测到细胞或抗体介导的免疫应答，并且在从移植物再培养的成纤维细胞中不能恢复表达。 这种现象让人想起在其他细胞类型中观察到的序列特异性基因失活。 由于外源蛋白的遗传操作和表达不影响移植细胞的存活，因此使用替代基因调控元件可能实现有效的长期治疗。
Genetically engineered fibroblasts have been successfully used to produce therapeutic proteins in animals, but sustained production of the proteins has not been achieved. This limits the potential of fibroblast-mediated gene therapy in humans. We have studied the phenomenon of decreased production in rats by using retroviral vectors carrying genes encoding human adenosine deaminase and neomycin phosphotransferase. While transplanted skin fibroblasts containing vector sequences persisted at constant levels for at least 8.5 mo, vector expression decreased by > 1500-fold after 1 mo. Cellular or antibody-mediated immune responses were not detected in transplanted animals, and expression could not be restored in fibroblasts recultivated from the grafts. This phenomenon is reminiscent of sequence-specific gene inactivation observed in other cell types. Because genetic manipulation and expression of foreign proteins did not affect survival of the transplanted cells, effective long-term therapy may be possible with the use of alternative gene regulatory elements.