Therapeutic strategies for inherited neurodegerative disorders using gene transfer and stem-cell transplantation technologies

1.基因治疗：我们研究了最近发现的硫酸酯酶激活酶（FGE）的表达芳基硫酸酯酶A（阿萨）的异染性脑白质营养不良（MLD）的基因治疗的效用。我们的数据表明，FGE共表达是必不可少的合成和分泌的功能阿萨在体外和体内。在治疗模型实验中，我们产生了携带阿萨或FGE cDNA的AAV 1载体。将这些载体同时注射到MLD小鼠的海马中，显著增加了阿萨活性，并降低了脑广泛区域中的硫苷脂积累。转棒试验和行走模式证实了行为的显著改善。这些结果支持FGE共表达对于有效合成功能性阿萨是必需的，并且对MLD基因治疗的开发具有重要意义。2.干细胞治疗：使用稳定重建的GFP转基因小鼠骨髓细胞的嵌合小鼠，我们证明了骨髓间充质干细胞（BSC）可以分化为多种细胞谱系。我们还成功地从脂肪组织（ASC）中制备了多能干细胞。用慢病毒载体稳定转染的神经前体细胞直接接种MLD，证实了细胞治疗MLD的可行性。基因工程干细胞可用于治疗神经退行性疾病。

1.Gene therapy : We examined the utility of recently identified sulfatase activating enzyme (FGE) for expression of arylsulfatase A(ASA) in gene therapy of metachromatic leukodustrophy(MLD). Our data demonstrated that FGE co-expression is essential for synthesis and secretion of functional ASA both in vitro and in vivo. In a therapeutic model experiment, we generated AAV1 vector carrying either ASA or FGE cDNA. Simultaneous injection of these vectors into the hippocampus of MLD mice significantly increased ASA activity and decreased sulfatide accumulation in the wide areas of the brain. Significant improvement of behavior was confirmed by the rotarod test and the walking pattern. These result support that FGE co-expression is essential for efficient synthesis of functional ASA and have significant implications for the development of MLD gene therapy.2.Stem cell therapy : Using chimeric mice stably reconstituted with bone marrow cells from GFP transgenic mice, we demonstrated that mesenchymal stem cells derived from bone marrow (BSC) could differentiate various cell lineages. We also succeeded in preparing pluripotent stem cells from adipose tissues(ASC). The feasibility of cell therapy of MLD was demonstrate by direct inoculation of neural progenitor cells stably tranduced with lentiviral vector carrying ASA cDNA. Genetically engineered stem cells may be useful for therapy for neurodegenerative disorders.