视网膜下间隙移植经基因修饰的视网膜色素上皮细胞对大鼠光感受器变性及视功能的保护作用

Subretinal transplantation of genetically modified human retinal pigment epithelium cells delay photoreceptor degeneration and functional deterioration in rats

目的 以RCS大鼠作模型,研究经基因修饰的永生化视网膜色素上皮细胞(RPE)视网膜下移植对光感受器变性的保护作用。方法 在绿色荧光蛋白基因逆转录病毒感染的基础上,利用脂质体介导节状神经生长因子(CNTF)表达质粒转移,修饰成人RPE细胞系CRL-2302。将1×105个表达绿色荧光蛋白(GFP)或GFP及CNTF的RPE细胞移植到4～5周龄RCS大鼠右眼视网膜下间隙,左眼不移植或注射。PBS作为对照。术后2、4、6、8、10和12周作荧光显微镜、光镜、电镜及电生理检查。结果 荧光显微镜观察,术后1周移植的人RPE细胞在RCS大鼠视网膜下间隙已扩散到几乎整个眼底,但随时间延长移植的细胞逐渐减少,术后6周仅残留少量移植细胞。光镜及电镜观察显示移植眼保留的光感受器数量明显较对照眼多,凋亡细胞则较对照跟少。此外,移植眼宿主RPE细胞形态较正常,并可见吞噬小体。视网膜电图(ERG)检查结果表明部分移植眼视网膜功能明显较对照眼好。结论 经过基因修饰的RPE细胞移植可延缓RCS大鼠视网膜光感受器变性,为治疗视网膜变性提供了新的途径。(中华眼科杂志,2004,40:552-556)

Objective To investigate whether subretinal transplantation of genetically modified human retinal pigment epithelium cells can rescue photoreceptor degeneration in RCS rats. Methods A spontaneously derived adult human retinal pigment epithelium ( RPE) cell line ( CRL-2302) was infected by gfp retrovirus, then transfected by liposome mediated CNTF expression plasmid transfer. Around 1 × 105 genetic modified cells were injected to subretinal space of the right eye at 4-5 weeks old, the left eye was left without injection or injected with PBS as controls. Results The GFP expression cells under the retina were observed from 1 to 6 weeks after transplantation. Histologic and ultrastructural assessment demonstrated substantial sparing of photoreceptors and correction of RPE phagocytosis defect. Electrophysiological assessment revealed an increased sensitivity of treated eyes to white light. Conclusion The results demonstrate that potential of genetically modified RPE cells for ultimate application in therapeutic transplantation strategies for human retinal degenerative diseases. (Chin J Ophthalmol, 2004, 40:552-556)