Regeneration of neurons and vessels using bone marrow cells and gene delivery

使用骨髓细胞和基因传递再生神经元和血管

• 批准号: 17300224
• 负责人: KUROIWA Toshihiko
• 金额: $ 3.84万
• 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 (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17300224/
• 关键词: Anti-aging; Senescence accelerated mouse; bone marrow cell; mesenchymal stromal cell; FGF; HGF; cerebral ischemia; 老化予防; 脳梗塞治療; 遺伝子導入; 認知症; 骨粗しょう症; 老化; 骨髄細胞移植; 脳萎縮; 老化促進マウス

We preliminarily intravenously implanted bone marrow cells from young mouse to old senescence accelerated mouse (SAM). We evaluate the memory function by water maze, however, no improvement in memory function was observed. Therefore, intravenous administration of young bone marrow has little effect on preventing aging and further enhanced treatment such as useful gene induction and targeting administration to the brain or bone marrow.Then 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 mesenchymal stromal cells (MSCs) by the herpes simplex virus type 1 (HSV-1) vector. 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-t … More reated 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. In conclusion these datum suggested 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.The hepatocyte growth factor (HGF) is also one of the useful gene for tissue restoration. we introduce a new strategy combining MSCs and ex vivo HGF gene transferring with a multimutated HSV-1 vector in a rat transient MCAO model. The significant difference of infarction areas on day was detected only between the MSC-HGF group and the PBS group with the superacute treatment, but was detected among each group on day 14 with both transplantations. After the superacute transplantation, we detected abundant expression of HGF protein in the ischemic brain of the MSC-HGF group compared with others on day 1 after treatment, and it was maintained for at least 2 weeks. Furthermore, we determined that the increased expression of HGF was derived from the transferred HGF gene in gene-modified MSCs. The percentage of apoptosis-positive cells in the ischemic boundary zone (IBZ) was significantly decreased, while that of remaining neurons in the cortex of the IBZ was significantly increased in the MSC-HGF group compared with others. The present study shows that combined therapy is more therapeutically efficient than MSC cell therapy alone, and it may extend the therapeutic time window from superacute to acute phase.Following these experiments, the improving effect is mainly due to anti-apoptotic and neuron-protective effects and there were no evidence for neuron-generation. Another effective methods such as intra-bone marrow injection of bone marrow cell should be examined in the future. Less

我们初步将年轻加速小鼠的骨髓细胞静脉注射到老年衰老小鼠（SAM）中，通过水迷宫评估记忆功能，但没有观察到记忆功能的改善，因此，静脉注射年轻骨髓对记忆功能影响不大。预防衰老并进一步加强治疗，如有用基因诱导和靶向给药至大脑或骨髓。然后，我们通过一种新的治疗策略检查了大鼠短暂性大脑中动脉闭塞（MCAO）后神经系统改善的效果通过 1 型单纯疱疹病毒 (HSV-1) 载体转染 FGF-2 基因的间充质基质细胞 (MSC)，接受 FGF-2 修饰的 MSC 的中风动物与其他组相比，在治疗 14 天后表现出显着的功能恢复。 MCAO，仅在 FGF-2 修饰的 MSC 治疗组中，FGF-2 修饰的 MSC-t 中的 FGF-2 产量显着减少……更多。 MCAO后3天和7天，与其他组相比，给予FGF-2修饰的MSC强烈表达FGF-2蛋白，这通过ELISA证明，这些数据表明FGF-2基因。 -与单独移植MSCs相比，用HSV-1载体修饰的MSCs有助于中风后显着的功能恢复。肝细胞生长因子（HGF）也是组织的有用基因之一我们在大鼠短暂性 MCAO 模型中引入了一种将 MSC 和离体 HGF 基因转移与多突变 HSV-1 载体相结合的新策略，仅在 MSC-HGF 组和 PBS 组之间检测到梗死面积的显着差异。与超急性治疗组相比，但在两次移植的第14天各组中均检测到超急性移植后，与MSC-HGF组相比，我们在缺血脑中检测到丰富的HGF蛋白表达。其他在治疗后第1天，并且维持至少2周此外，我们确定HGF表达的增加源自基因修饰的MSC中转移的HGF基因的凋亡阳性细胞的百分比。与其他组相比，MSC-HGF组的边界区（IBZ）显着减少，而IBZ皮质中剩余神经元的数量显着增加，本研究表明联合治疗比单独的MSC细胞治疗更有效。和它可以将治疗时间窗口从超急性期延长到急性期。根据这些实验，改善效果主要是由于抗凋亡和神经元保护作用，并且没有证据表明骨髓内治疗等其他有效方法。以后应少注射骨髓细胞进行检查。

项目成果

Novel Therapeutic Strategy for Stroke in Rats by Bone Marrow Stromal Cells and ex vivo HGF Gene Transfer with HSV-1 Vector

• DOI: 10.1038/sj.jcbfm.9600273
• 发表时间: 2006-09
• 期刊: Journal of Cerebral Blood Flow & Metabolism
• 影响因子: 6.3
• 作者: Ming-Zhu Zhao;N. Nonoguchi;N. Ikeda;Takuji Watanabe;D. Furutama;Daisuke Miyazawa;H. Funakoshi