An approach for ALS genetic therapy using Herpes Simplex Virus Vector

使用单纯疱疹病毒载体进行 ALS 基因治疗的方法

• 批准号: 15590916
• 负责人: KIMURA Fumiharu
• 金额: $ 1.98万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590916/
• 关键词: ALS; FGF2; MCAO; one marrow stromal cell; HSV-1 vector; 遺伝子治療; HGF; 骨髄間葉系幹細胞; 脳虚血; 筋萎縮性側策硬化症; 単純ヘルペス; 肝細胞増殖因子; 線維芽細胞増殖因子; 血管内皮細胞増殖因子; ウイルスベクター; Herpes Vector; Motor Neuron disease

Background and Purpose : Fibroblast growth factor-2 (FGF-2) administration and bone marrow stromal cell (MSC) transplantation could improve neurological deficits after occlusive cerebrovascular disease. In the present study, we examined the effects of neurological improvement after transient middle cerebral artery occlusion (MCAO) in rats by a novel therapeutic strategy with FGF-2 gene transferred MSCs by the herpes simplex virus type 1 (HSV-1) vector. Methods Adult Wistar rats were anesthetized. Nonmodified MSCs, FGF-2 modified MSCs with HSV-1 1764/-4/pR19/ ssIL2-FGF-2, or PBS was administered intracerebrally 24 hours after transient right MCAO. All animals underwent behavioral tests for 21 days, and the infarction volume with 2-3-5-triphenylterazolium was detected 3 days and 14 days after the MCAO. Three days and 7 days after the MCAO, the FGF-2 production in the ipsilateral hemisphere of the MCAO was measured with ELISA. Seven and 14 days after the MCAO, immunohistochemical staining for FGF-2 was applied. Results The stroke animals receiving FGF-2 modified MSCs demonstrated significant functional recovery compared with the other groups. Fourteen days after the MCAO, there was a significant reduction in infarction volume only in FGF-2 modified MSC-treated group. FGF-2 production in the FGF-2 modified MSC-treated brain was significantly higher compared with the other groups at 3 and 7 days after MCAO. Administrated FGF-2 modified MSCs strongly expressed the FGF-2 protein, which was proven by ELISA. Conclusions Our data suggest that the FGF-2 gene modified MSCs with the HSV-1 vector can contribute to remarkable functional recovery after stroke compared with MSCs transplantation alone.

背景和目的：成纤维细胞生长因子-2（FGF-2）和骨髓基质细胞（MSC）移植可改善闭塞性脑血管病后的神经功能缺损。在本研究中，我们研究了一种新的治疗策略与FGF-2基因转染的单纯疱疹病毒1型（HSV-1）载体的MSC短暂大脑中动脉闭塞（MCAO）后，大鼠神经功能的改善效果。方法成年Wistar大鼠麻醉。在短暂性右侧MCAO后24小时，脑内施用未修饰的MSC、用HSV-11764/-4/pR19/ssIL 2-FGF-2修饰的MSC或PBS。所有动物进行21天的行为学测试，并在MCAO后3天和14天用2-3-5-三苯基四唑检测梗死体积。在MCAO后3天和7天，用ELISA测量MCAO同侧半球中的FGF-2产生。在MCAO后7天和14天，应用FGF-2免疫组织化学染色。结果与其他组相比，接受FGF-2修饰的MSCs的中风动物表现出显著的功能恢复。MCAO后14天，仅FGF-2修饰的MSC治疗组的梗死体积显著减少。在MCAO后第3天和第7天，FGF-2修饰的MSC处理的脑中FGF-2的产生显著高于其他组。经FGF-2修饰的MSCs表达FGF-2蛋白，ELISA检测证实其表达增强。结论FGF-2基因修饰的MSCs经HSV-1载体转染后，与单纯MSCs移植相比，可促进脑卒中后功能的恢复。

项目成果

Wheelchair economy class syndrome in amyotrophic lateral sclerosis

• DOI: 10.1016/j.nmd.2005.12.006
• 发表时间: 2006-03-01
• 期刊: NEUROMUSCULAR DISORDERS
• 影响因子: 2.8
• 作者: Kimura, F;Ishida, S;Hanafusa, T
• 通讯作者: Hanafusa, T

Post-lschemic Intraventricular Administration of FGF-2 Expressing Adenoviral Vectors Improves Neurological Outcome and Reduces Infarct Volume After Transient Focal Cerebral Ischemia in Rats.

缺血后脑室内注射表达 FGF-2 的腺病毒载体可改善大鼠短暂性局灶性脑缺血后的神经学结果并减少梗塞体积。

• 发表时间: 2004
• 期刊: J Cerebral Blood Flow and Metabolism 24
• 作者: Keiichi Ishihara;et al.;Watanabe-T et al.
• 通讯作者: Watanabe-T et al.

Postischemic- intraventricular administration of FGF-2 expressing adenobiral bettors improves neurologic outcome and reduces infarct volume after transient focal cerebral ischemia in rats.

缺血后脑室内给予表达 FGF-2 的腺苷酸下注剂可改善大鼠短暂性局灶性脑缺血后的神经系统结果并减少梗塞体积。

• 发表时间: 2004
• 期刊: J Cereb Blood Flow Metab 24(11)
• 作者: Watanabe T;Okuda Y;Nonoguchi N;Zhao MZ;Kajimoto Y;Furutama D;Yukawa H;Shibata MA;Otsuki Y;Kuroiwa T;Miyatake
• 通讯作者: Miyatake

Post-Ischemic Intraventricular Administration of FGF-2 Expressing Adenoviral Vectors Improves Neurological Outcome and Reduces Infarct Volume After Transient Focal Cerebral Ischemia in Rats.

缺血后脑室内注射表达 FGF-2 的腺病毒载体可改善大鼠短暂性局灶性脑缺血后的神经学结果并减少梗塞体积。

• 发表时间: 2004
• 期刊: J Cerebral Blood Flow and Metabolism 24
• 作者: F.Kimura;C.Fujimura;S.Ishida;H.Nakajima;D.Furutama;H.Uehara;K.Shinoda;M.Sugino;T.Hanafusa;Kimuea F et al.;Watanabe T et al.
• 通讯作者: Watanabe T et al.

木村 文治: "筋萎縮性側索硬化症100例の変遷"臨床神経. 43. 385-391 (2003)

Fumiharu Kimura：“100 例肌萎缩侧索硬化症的变化”《临床神经病学》43. 385-391 (2003)。