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

利用基因转移和干细胞移植技术治疗遗传性神经退行性疾病的策略

• 批准号: 14370253
• 负责人: SHIMADA Takashi
• 金额: $ 8.96万
• 依托单位: Nippon Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370253/
• 关键词: gene therapy; cell therapy; lysosomal disorders; stem cells; neural stem cells; AAV vector; 異染性ロイコジストロフィー; アリルサルファターゼ; 多能性幹細胞; 脂肪組織由来幹細胞; HIVベクター; 異染性白質ジストロフィー; 多能性骨髄幹細胞; gene therapy; cell therapy; lysosomal disorders; stem cells; neural stem cells; AAV vector; 異染性ロイコジストロフィー; アリルサルファターゼ; 多能性幹細胞; 脂肪組織由来幹細胞; HIVベクター; 異染性白質ジストロフィー; 多能性骨髄幹細胞

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.

1.基因疗法：我们检查了最近确定的硫酸酶激活酶（FGE）的效用，以表达芳基硫酸酯酶A（ASA）（ASA）在实质性白细胞植物（MLD）基因治疗中的表达。我们的数据表明，FGE共表达对于在体外和体内的功能ASA的合成和分泌至关重要。在治疗模型实验中，我们生成了带有ASA或FGE cDNA的AAV1载体。同时将这些载体注射到MLD小鼠的海马中，显着增加了ASA活性，并减少了大脑广泛区域的硫化物积累。通过Rotarod检验和步行模式证实了行为的显着改善。这些结果支持FGE共表达对于有效合成功能性ASA并对MLD基因治疗的发展具有重要意义。我们还成功地从脂肪组织（ASC）制备了多能干细胞。通过直接接种携带ASA cDNA的慢病毒载体稳定地将MLD细胞治疗的可行性证明了。基因设计的干细胞可能对神经退行性疾病的治疗有用。