Ohsugi K, Kobayashi K, Itoh K, Sakuraba H, Sakuragawa N
Department of Inherited Metabolic Disease, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan.
J Hum Genet. 2000;45(1):1-5. doi: 10.1007/s100380050001.
Fabry disease is an X-linked inherited metabolic disorder caused by a deficiency of alpha-galactosidase (alpha-gal), resulting in the accumulation of ceramide trihexoside (CTH) in body fluids and in many organs and tissues. We constructed a recombinant adenovirus with a human alpha-gal cDNA (AxCAG alpha-gal), and transfected this vector to skin fibroblasts from Fabry patients. Transfected cells expressed high amounts of alpha-gal in their cytoplasm, and a high level of alpha-gal activity was detected in the medium. The accumulated CTH in the fibroblasts disappeared 3 days after infection. The secreted alpha-gal also eliminated the accumulated CTH from uninfected patient's cells. The enzyme may be taken up through mannose-6-phosphate receptors, as the addition of mannose-6-phosphate to the medium completely inhibited the uptake of the enzyme. The infected cells continued to express alpha-gal for more than 10 days. These results suggest that AxCAG alpha-gal could be used as enzyme replacement gene therapy for Fabry disease.
法布里病是一种X连锁遗传性代谢紊乱疾病，由α-半乳糖苷酶（α-gal）缺乏引起，导致神经酰胺三己糖苷（CTH）在体液以及许多器官和组织中蓄积。我们构建了携带人α-gal cDNA的重组腺病毒（AxCAG α-gal），并将该载体转染至法布里病患者的皮肤成纤维细胞。转染后的细胞在其细胞质中大量表达α-gal，并且在培养基中检测到高水平的α-gal活性。成纤维细胞中蓄积的CTH在感染后3天消失。分泌的α-gal也消除了未感染患者细胞中蓄积的CTH。该酶可能通过甘露糖-6-磷酸受体被摄取, 因为向培养基中添加甘露糖-6-磷酸完全抑制了该酶的摄取。感染的细胞持续表达α-gal超过10天。这些结果表明，AxCAG α-gal可作为法布里病的酶替代基因疗法。
