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磺化酶激活酶（FGE）在异色性脑白质症（MLD）基因治疗中表达芳基磺化酶A（ASA）的效用。我们的数据表明，FGE共表达对于体外和体内功能性ASA的合成和分泌至关重要。在治疗模型实验中，我们生成了携带ASA或FGE cDNA的AAV1载体。将这些载体同时注射到MLD小鼠的海马体中，可显著提高ASA活性，减少脑宽区域的硫脂质积累。通过旋转杆测试和行走方式证实了行为的显著改善。这些结果支持FGE共表达是高效合成功能性ASA的必要条件，并对MLD基因治疗的发展具有重要意义。干细胞治疗：利用从GFP转基因小鼠获得的骨髓细胞稳定重组的嵌合小鼠，我们证明了来自骨髓的间充质干细胞（BSC）可以分化各种细胞系。我们还成功地从脂肪组织（ASC）中制备了多能干细胞。通过携带ASA cDNA的慢病毒载体稳定诱导神经祖细胞直接接种，证实了细胞治疗MLD的可行性。基因工程干细胞可能对神经退行性疾病的治疗有用。

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.