NT-3 expression from engineered olfactory ensheathing glia promotes spinal sparing and regeneration.
NT-3 expression from engineered olfactory ensheathing glia promotes spinal sparing and regeneration.
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DOI：
10.1093/brain/awh424
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发表时间：
2005-04
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通讯作者：
M. Ruitenberg;Dane B Levison;Seok Voon Lee;J. Verhaagen;A. Harvey;G. Plant
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作者：
M. Ruitenberg;Dane B Levison;Seok Voon Lee;J. Verhaagen;A. Harvey;G. Plant
利用腺病毒（AdV）载体编码神经营养因子-3 （AdV- nt -3）或细菌标记酶-半乳糖苷酶（LacZ基因）介导嗅鞘胶质细胞（OEG）培养。原位杂交和酶联免疫吸附测定技术显示，AdV载体转导的OEG表达了高水平的重组神经营养蛋白。载体衍生的NT-3的生物活性是在背根神经节神经突生长试验中确定的。在单侧颈（C4）皮质脊髓束（CST）横断后立即将工程细胞悬液注射到成年Fischer 344大鼠脊髓中。移植的动物总共接受了20万个细胞；未转导的脑电图或分别用编码NT-3或LacZ的AdV载体转导的脑电图。损伤后3个月，与对照（中剂量注射）大鼠相比，所有oeg移植大鼠的病变体积都明显变小。来自对侧感觉运动皮层的损伤CST投影的顺行追踪显示，仅在移植了oeg - nt -3的大鼠中，远端CST轴突的数量显著增加。行为学分析采用开阔场地运动评分法和Eshkol-Wachman动作记数法对所有大鼠进行前肢到达任务。行为测试分析显示，实验组之间的恢复没有显著差异，尽管运动分析表明，可能的代偿机制在脑电图植入后发生。结果表明，OEG移植本身可以促进损伤后的组织保留，但是，经过适当的基因修饰，这些嗅觉来源的细胞在促进受损成人CST轴突的远距离维持/再生方面变得更加有效。
Adenoviral (AdV) vectors encoding neurotrophin-3 (AdV-NT-3) or the bacterial marker enzyme beta-galactosidase (LacZ gene) were used to transduce olfactory ensheathing glia (OEG) cultures. AdV vector-transduced OEG expressed high levels of recombinant neurotrophin as shown by in situ hybridization and enzyme-linked immunosorbent assay techniques. The biological activity of vector-derived NT-3 was determined in a dorsal root ganglia neurite outgrowth assay. Engineered cell suspensions were then injected into adult Fischer 344 rat spinal cord immediately after unilateral cervical (C4) corticospinal tract (CST) transection. Transplanted animals received a total of 200,000 cells; either non-transduced OEG or OEG transduced with AdV vectors encoding NT-3 or LacZ, respectively. At 3 months after injury, lesion volumes were significantly smaller in all OEG-transplanted rats when compared with control (medium-injected) rats. Anterograde tracing of the lesioned CST projection, originating from the contralateral sensorimotor cortex, showed a significantly greater number of distal CST axons only in OEG-NT-3-transplanted rats. Behavioural analysis was performed on all rats using open field locomotion scoring, and a forelimb reaching task with Eshkol-Wachman movement notation. Analysis of behavioural tests revealed no significant differences in recovery between experimental groups, although movement analysis indicated that possible compensatory mechanisms were occurring after OEG implantation. The results demonstrate that OEG transplantation per se can promote tissue sparing after injury, but, after appropriate genetic modification, these olfactory-derived cells become far more effective in promoting long-distance maintenance/regeneration of lesioned adult CST axons.