Wolfe J H, Schuchman E H, Stramm L E, Concaugh E A, Haskins M E, Aguirre G D, Patterson D F, Desnick R J, Gilboa E
Laboratory of Pathology, University of Pennsylvania School of Veterinary Medicine, Philadelphia 19104.
Proc Natl Acad Sci U S A. 1990 Apr;87(8):2877-81. doi: 10.1073/pnas.87.8.2877.
Retroviral vectors were constructed containing a rat beta-glucuronidase cDNA driven by heterologous promoters. Vector-mediated gene transfer into human and canine beta-glucuronidase-deficient mucopolysaccharidosis type VII fibroblasts completely corrected the deficiency in beta-glucuronidase enzymatic activity. In primary cultures of canine mucopolysaccharidosis type VII retinal pigment epithelial cells, which contain large amounts of undegraded glycosaminoglycan substrates, vector correction restored normal processing of specific glycosaminoglycans in the lysosomal compartment. In canine mucopolysaccharidosis type VII bone marrow cells, beta-glucuronidase was expressed at high levels in transduced cells. Thus, the vector-encoded beta-glucuronidase was expressed at therapeutic levels in the appropriate organelle and corrected the metabolic defect in cells exhibiting the characteristic pathology of this lysosomal storage disorder.
构建了逆转录病毒载体,其包含由异源启动子驱动的大鼠β-葡萄糖醛酸酶cDNA。载体介导的基因转移到人类和犬类β-葡萄糖醛酸酶缺陷的VII型粘多糖贮积症成纤维细胞中,完全纠正了β-葡萄糖醛酸酶酶活性的缺陷。在犬类VII型粘多糖贮积症视网膜色素上皮细胞的原代培养物中,其含有大量未降解的糖胺聚糖底物,载体校正恢复了溶酶体区室中特定糖胺聚糖的正常加工。在犬类VII型粘多糖贮积症骨髓细胞中,β-葡萄糖醛酸酶在转导细胞中高水平表达。因此,载体编码的β-葡萄糖醛酸酶在适当的细胞器中以治疗水平表达,并纠正了表现出这种溶酶体贮积症特征性病理的细胞中的代谢缺陷。