Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, 21941-901, Brazil.
Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, FL, 32610, USA.
Gene Ther. 2020 Feb;27(1-2):27-39. doi: 10.1038/s41434-019-0089-0. Epub 2019 Jun 26.
After an injury, axons in the central nervous system do not regenerate over large distances and permanently lose their connections to the brain. Two promising approaches to correct this condition are cell and gene therapies. In the present work, we evaluated the neuroprotective and neuroregenerative potential of pigment epithelium-derived factor (PEDF) gene therapy alone and combined with human mesenchymal stem cell (hMSC) therapy after optic nerve injury by analysis of retinal ganglion cell survival and axonal outgrowth. Overexpression of PEDF by intravitreal delivery of AAV2 vector significantly increased Tuj1-positive cells survival and modulated FGF-2, IL-1ß, Iba-1, and GFAP immunostaining in the ganglion cell layer (GCL) at 4 weeks after optic nerve crush, although it could not promote axonal outgrowth. The combination of AAV2.PEDF and hMSC therapy showed a higher number of Tuj1-positive cells and a pronounced axonal outgrowth than unimodal therapy after optic nerve crush. In summary, our results highlight a synergistic effect of combined gene and cell therapy relevant for future therapeutic interventions regarding optic nerve injury.
在损伤后，中枢神经系统中的轴突不能在长距离上再生，并永久失去与大脑的连接。两种有前途的纠正这种情况的方法是细胞和基因治疗。在本工作中，我们通过分析视网膜神经节细胞存活和轴突生长来评估单独的色素上皮衍生因子（PEDF）基因治疗和联合人骨髓间充质干细胞（hMSC）治疗对视神经损伤的神经保护和神经再生潜力。通过玻璃体内注射 AAV2 载体过表达 PEDF，可显著增加视神经挤压后 4 周时神经节细胞层（GCL）中 Tuj1 阳性细胞的存活，并调节 FGF-2、IL-1β、Iba-1 和 GFAP 的免疫染色，但不能促进轴突生长。AAV2.PEDF 和 hMSC 联合治疗在视神经挤压后显示出比单一治疗更高数量的 Tuj1 阳性细胞和更明显的轴突生长。总之，我们的结果突出了联合基因和细胞治疗的协同作用，这对于视神经损伤的未来治疗干预具有重要意义。
