Higuchi Koji, Yoshimitsu Makoto, Fan Xin, Guo Xiaoxin, Rasaiah Vanessa I, Yen Jennifer, Tei Chuwa, Takenaka Toshihiro, Medin Jeffrey A
University Health Network (UHN), Toronto, Ontario, Canada.
Mol Med. 2010 May-Jun;16(5-6):216-21. doi: 10.2119/molmed.2009.00163. Epub 2010 Feb 17.
The protein transduction domain from human immunodeficiency virus (HIV) Tat allows proteins to penetrate the cell membrane. Enhanced cellular uptake of therapeutic proteins could benefit a number of disorders. This is especially true for lysosomal storage disorders (LSDs) where enzyme replacement therapy (ERT) and gene therapy have been developed. We developed a novel recombinant lentiviral vector (LV) that engineers expression of alpha-galactosidase A (alpha-gal A)-Tat fusion protein for correction of Fabry disease, the second-most prevalent LSD with manifestations in the brain, kidney and heart. In vitro experiments confirmed mannose-6-phosphate independent uptake of the fusion factor. Next, concentrated therapeutic LV was injected into neonatal Fabry mice. Analysis of tissues at 26 wks demonstrated similar alpha-gal A enzyme activities but enhanced globotriaosylceramide (Gb3) reduction in hearts and kidneys compared with the alpha-gal A LV control. This strategy might advance not only gene therapy for Fabry disease and other LSDs, but also ERT, especially for cardiac Fabry disease.
来自人类免疫缺陷病毒（HIV）Tat的蛋白质转导结构域可使蛋白质穿透细胞膜。增强治疗性蛋白质的细胞摄取可能对多种疾病有益。对于已开发出酶替代疗法（ERT）和基因疗法的溶酶体贮积症（LSD）来说尤其如此。我们开发了一种新型重组慢病毒载体（LV），该载体可设计表达α-半乳糖苷酶A（α-gal A）-Tat融合蛋白，用于纠正法布里病，这是第二常见的LSD，会在脑、肾和心脏出现症状。体外实验证实了融合因子的非磷酸甘露糖依赖性摄取。接下来，将浓缩的治疗性LV注射到新生法布里小鼠体内。26周时对组织的分析表明，与α-gal A LV对照组相比，心脏和肾脏中的α-gal A酶活性相似，但球三糖神经酰胺（Gb3）的减少更为明显。这种策略不仅可能推动法布里病和其他LSD的基因治疗，还可能推动ERT，尤其是针对心脏法布里病。
