Johnen Sandra, Djalali-Talab Yassin, Kazanskaya Olga, Möller Theresa, Harmening Nina, Kropp Martina, Izsvák Zsuzsanna, Walter Peter, Thumann Gabriele
Department of Ophthalmology, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.
Department of Ophthalmology, University Hospitals of Geneva, Street Alcide-Jentzer 22, 1211 Geneva 14, Switzerland.
Biomed Res Int. 2015;2015:863845. doi: 10.1155/2015/863845. Epub 2015 Dec 1.
Pigment epithelium-derived factor (PEDF) is a potent multifunctional protein that inhibits angiogenesis and has neurogenic and neuroprotective properties. Since the wet form of age-related macular degeneration is characterized by choroidal neovascularization (CNV), PEDF would be an ideal candidate to inhibit CNV and support retinal pigment epithelial (RPE) cells. However, its short half-life has precluded its clinical use. To deliver PEDF to the subretinal space, we transfected RPE cells with the PEDF gene using the Sleeping Beauty transposon system. Transfected cells expressed and secreted biologically active recombinant PEDF (rPEDF). In cultures of human umbilical vein endothelial cells, rPEDF reduced VEGF-induced cumulative sprouting by ≥47%, decreased migration by 77%, and increased rate of apoptosis at least 3.4 times. rPEDF induced neurite outgrowth in neuroblastoma cells and protected ganglion and photoreceptor cells in organotypic retinal cultures. In a rat model of CNV, subretinal transplantation of PEDF-transfected cells led to a reduction of the CNV area by 48% 14 days after transplantation and decreased clinical significant lesions by 55% and 40% after 7 and 14 days, respectively. We showed that transplantation of pigment epithelial cells overexpressing PEDF can restore a permissive subretinal environment for RPE and photoreceptor maintenance, while inhibiting choroidal blood vessel growth.
色素上皮衍生因子（PEDF）是一种强大的多功能蛋白质，具有抑制血管生成以及神经生成和神经保护特性。由于湿性年龄相关性黄斑变性的特征是脉络膜新生血管（CNV），PEDF将是抑制CNV和支持视网膜色素上皮（RPE）细胞的理想候选物。然而，其半衰期短阻碍了其临床应用。为了将PEDF递送至视网膜下间隙，我们使用睡美人转座子系统用PEDF基因转染RPE细胞。转染的细胞表达并分泌具有生物活性的重组PEDF（rPEDF）。在人脐静脉内皮细胞培养物中，rPEDF使VEGF诱导的累积芽生减少≥47%，迁移减少77%，凋亡率至少增加3.4倍。rPEDF在神经母细胞瘤细胞中诱导神经突生长，并在视网膜组织型培养物中保护神经节和光感受器细胞。在CNV大鼠模型中，PEDF转染细胞的视网膜下移植导致移植后14天CNV面积减少48%，7天和14天后临床显著病变分别减少55%和40%。我们表明，过表达PEDF的色素上皮细胞移植可以恢复有利于RPE和光感受器维持的视网膜下环境，同时抑制脉络膜血管生长。
