Cross-correction of beta-glucuronidase deficiency by retroviral vector-mediated gene transfer.
Cross-correction of beta-glucuronidase deficiency by retroviral vector-mediated gene transfer.
复制标题
通过逆转录病毒载体介导的基因转移交叉校正β-葡萄糖醛酸酶缺陷。
DOI：
10.1006/excr.1994.1298
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发表时间：
1994
影响因子：
3.7
通讯作者：
Wolfe,JH
中科院分区：
文献类型：
--
作者：
Taylor,RM;Wolfe,JH
从基因修饰细胞中交叉校正溶酶体贮积障碍的肌蛋白体外模型被开发出来，以接近体内基因治疗所需的生理条件。研究了β-葡萄糖醛酸酶（GUSB）缺陷粘多糖病（MPS） VII型（Sly病）细胞，以确定酶转移的量和稳定性。利用编码大鼠或人GUSB的逆转录病毒载体进行基因转移，可以纠正人、狗和小鼠培养的MPS VII成纤维细胞的酶缺陷。载体转导的细胞将GUSB释放到培养上清中，其数量与细胞GUSB活性的数量成正比。输出的GUSB在组织培养基中活性稳定至少两周。当载体感染的供体细胞与缺陷靶细胞通过透液膜分离时，未感染的MPS VII靶细胞加入了培养基中的GUSB。去除酶供体细胞后，交叉校正靶细胞中的高水平GUSB迅速下降。
Anin vitromodel for cross-correction of lysosomal storage disorders from genetically modified cells was developed to approximate the physiological conditions needed for gene therapyin vivo. β-Glucuronidase (GUSB)-deficient mucopolysaccharidosis (MPS) type VII (Sly disease) cells were studied to determine the amount and stability of enzyme transfer. Gene transfer by retroviral vectors encoding rat or human GUSB corrected the enzymatic deficiency in cultured MPS VII fibroblasts from humans, dogs, and mice. The vector-transduced cells released GUSB into the culture supernatant in amounts proportional to the amounts of cellular GUSB activity. The activity of the exported GUSB was stable in tissue culture medium for at least two weeks. Uninfected MPS VII target cells incorporated GUSB from the medium when the vector-infected donor cells were separated from the deficient target cells by a fluid-permeable membrane. The high level of GUSB in the cross-corrected target cells decreased rapidly after the enzyme donor cells were removed.