Correction in trans for Fabry disease: Expression, secretion, and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector
Correction in trans for Fabry disease: Expression, secretion, and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector
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DOI：
10.1073/pnas.93.15.7917
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发表时间：
1996-07-23
影响因子：
11.1
通讯作者：
Brady, RO
中科院分区：
文献类型：
--
作者：
Medin, JA;Tudor, M;Brady, RO
法布里病是一种由α -半乳糖苷酶a （α -半乳糖苷酶a； EC 3.2.1.22）缺乏引起的x连锁代谢紊乱。患者积累的鞘糖脂具有末端α -半乳糖残基，这些残基来自细胞内合成、循环代谢产物或衰老细胞的生物降解。病人最终死于肾脏、心脏或脑血管疾病。没有具体的治疗方法。改善这种疾病的一种可能的方法是针对循环造血细胞进行纠正性基因转移治疗。为此，研究人员开发了一种两性性病毒产生细胞系，该细胞系可产生高滴度（bbb10 (6) ipp / ml）的重组逆转录病毒，用于转导和校正靶细胞。病毒产生细胞也表现出细胞内和分泌的α -半乳糖a的大量表达。为了检验这种治疗载体的效用，我们用重组病毒感染法布里患者的皮肤成纤维细胞，纠正了代谢缺陷，并观察了分泌的酶。此外，分泌的酶被发现以甘露糖-6-磷酸受体依赖的方式被未校正的细胞吸收。在相关实验中，来自Fabry患者的永生化B细胞系，作为血液学传递测试系统，被转导。与成纤维细胞一样，转导的患者B细胞系表现出内源性酶纠正和少量分泌以及未纠正细胞的摄取。这些研究表明，转导细胞的内源性代谢纠正与分泌相结合，可以为未修饰的患者旁观者细胞提供持续的纠正材料来源（代谢协同性）。
Fabry disease is an X-linked metabolic disorder due to a deficiency of alpha-galactosidase A (alpha-gal A; EC 3.2.1.22). Patients accumulate glycosphingolipids with terminal alpha-galactosyl residues that come from intracellular synthesis, circulating metabolites, or from the biodegradation of senescent cells. Patients eventually succumb to renal, cardio-, or cerebrovascular disease. No specific therapy exists. One possible approach to ameliorating this disorder is to target corrective gene transfer therapy to circulating hematopoietic cells. Toward this end, an amphotropic virus-producer cell line has been developed that produces a high titer (> 10(6) i.p. per ml) recombinant retrovirus constructed to transduce and correct target cells. Virus-producer cells also demonstrate expression of large amounts of both intracellular and secreted alpha-gal A. To examine the utility of this therapeutic vector, skin fibroblasts from Fabry patients were corrected for the metabolic defect by infection with this recombinant virus and secreted enzyme was observed. Furthermore, the secreted enzyme was found to be taken up by uncorrected cells in a mannose-6-phosphate receptor-dependent manner. In related experiments, immortalized B cell lines from Fabry patients, created as a hematologic delivery test system, were transduced. As with the fibroblasts, transduced patient B cell lines demonstrated both endogenous enzyme correction and a small amount of secretion together with uptake by uncorrected cells. These studies demonstrate that endogenous metabolic correction in transduced cells, combined with secretion, may provide a continuous source of corrective material in trans to unmodified patient bystander cells (metabolic cooperativity).