University Health Network, Toronto, Ontario, Canada.
Mol Ther. 2012 Jul;20(7):1454-61. doi: 10.1038/mt.2012.64. Epub 2012 Apr 3.
Fabry disease is a lysosomal storage disorder caused by a deficiency of α-galactosidase A (α-gal A) activity that results in progressive globotriaosylceramide (Gb(3)) deposition. We created a fully congenic nonobese diabetic (NOD)/severe combined immunodeficiency (SCID)/Fabry murine line to facilitate the in vivo assessment of human cell-directed therapies for Fabry disease. This pure line was generated after 11 generations of backcrosses and was found, as expected, to have a reduced immune compartment and background α-gal A activity. Next, we transplanted normal human CD34(+) cells transduced with a control (lentiviral vector-enhanced green fluorescent protein (LV-eGFP)) or a therapeutic bicistronic LV (LV-α-gal A/internal ribosome entry site (IRES)/hCD25). While both experimental groups showed similar engraftment levels, only the therapeutic group displayed a significant increase in plasma α-gal A activity. Gb(3) quantification at 12 weeks revealed metabolic correction in the spleen, lung, and liver for both groups. Importantly, only in the therapeutically-transduced cohort was a significant Gb(3) reduction found in the heart and kidney, key target organs for the amelioration of Fabry disease in humans.
法布瑞氏病是一种溶酶体贮积病,由α-半乳糖苷酶 A(α-gal A)活性缺乏引起,导致进行性糖鞘脂(Gb(3))沉积。我们构建了一个完全同基因非肥胖型糖尿病(NOD)/严重联合免疫缺陷(SCID)/法布瑞氏病小鼠系,以促进法布瑞氏病的人类细胞定向治疗的体内评估。这条纯系经过 11 代回交后产生,正如预期的那样,其免疫成分和背景α-gal A 活性减少。接下来,我们移植了经慢病毒载体增强型绿色荧光蛋白(LV-eGFP)或治疗性双顺反子慢病毒(LV-α-gal A/内部核糖体进入位点(IRES)/hCD25)转导的正常人类 CD34(+)细胞。虽然两个实验组的植入水平相似,但只有治疗组的血浆α-gal A 活性显著增加。12 周时 Gb(3)的定量分析显示,两组的脾脏、肺和肝脏都有代谢纠正。重要的是,只有在经治疗转导的队列中,心脏和肾脏中的 Gb(3)才显著减少,这是法布瑞氏病在人类中改善的关键靶器官。