Yoshimitsu M, Higuchi K, Ramsubir S, Nonaka T, Rasaiah V I, Siatskas C, Liang S-B, Murray G J, Brady R O, Medin J A
Division of Stem Cell and Developmental Biology, Ontario Cancer Institute, University Health Network, Toronto, Ontario, Canada.
Gene Ther. 2007 Feb;14(3):256-65. doi: 10.1038/sj.gt.3302839. Epub 2006 Aug 24.
A deficiency in alpha-galactosidase A (alpha-gal A) activity causes Fabry disease. Virus-based delivery of genes can correct cells and establish a sustained supply of therapeutic proteins. Recombinant lentiviral vectors (LVs) show promise in this context. We first demonstrate LV-mediated marking of peripheral blood (PB) cells by transduction/transplantation of hematopoietic stem/progenitor cells. Stable enGFP expression was observed in PB for 37 weeks. Next, we transplanted Fabry mice with bone marrow mononuclear cells (BMMNCs) transduced a single time with a LV encoding the human alpha-gal A cDNA. Sustained expression of functional alpha-gal A in Fabry mice was observed over 24 weeks. Plasma alpha-gal A activity from treated Fabry mice was two-fold higher than wild-type controls. Increased alpha-gal A activity, often to supra-normal levels, and reduction of globotriaosylceramide, a glycolipid that accumulates in Fabry disease, was observed in all organs assessed. In secondary bone marrow transplantations, Fabry mice showed multilineage marking of PB, splenocytes and BMMNCs, along with therapeutic levels of alpha-gal A activity in plasma and organs over 20 weeks. Lastly, we transduced mobilized PB CD34(+) cells from a Fabry patient and observed corresponding enzymatic increases. Thus a single LV-mediated transduction of primitive hematopoietic cells can result in sustained correction for Fabry disease.
α-半乳糖苷酶A（α-gal A）活性缺乏会导致法布里病。基于病毒的基因递送可以纠正细胞并建立治疗性蛋白质的持续供应。在这种情况下，重组慢病毒载体（LVs）显示出前景。我们首先通过造血干/祖细胞的转导/移植证明了LV介导的外周血（PB）细胞标记。在PB中观察到稳定的增强型绿色荧光蛋白（enGFP）表达持续37周。接下来，我们用编码人α-gal A cDNA的LV单次转导的骨髓单个核细胞（BMMNCs）移植法布里病小鼠。在24周内观察到法布里病小鼠中功能性α-gal A的持续表达。治疗后的法布里病小鼠血浆α-gal A活性比野生型对照高两倍。在所有评估的器官中均观察到α-gal A活性增加，常常达到超正常水平，并且法布里病中积累的糖脂神经酰胺三己糖苷减少。在二次骨髓移植中，法布里病小鼠在20周内显示出PB、脾细胞和BMMNCs的多谱系标记，以及血浆和器官中α-gal A活性的治疗水平。最后，我们转导了一名法布里病患者动员的PB CD34(+)细胞，并观察到相应的酶活性增加。因此，原始造血细胞的单次LV介导的转导可以导致法布里病的持续纠正。
