Department of Ophthalmology, University Hospital RWTH Aachen, 52074 Aachen, Germany.
Division of Medical Biotechnology, Paul-Ehrlich-Institute, 63225 Langen, Germany.
Int J Mol Sci. 2022 Oct 26;23(21):12982. doi: 10.3390/ijms232112982.
More and more patients suffer from multifactorial neurodegenerative diseases, such as age-related macular degeneration (AMD). However, their pathological mechanisms are still poorly understood, which complicates the development of effective therapies. To improve treatment of multifactorial diseases, cell-based gene therapy can be used to increase the expression of therapeutic factors. To date, there is no approved therapy for dry AMD, including late-stage geographic atrophy. We present a treatment option for dry AMD that transfers the brain-derived neurotrophic factor () gene into retinal pigment epithelial (RPE) cells by electroporation using the plasmid-based () transposon system. ARPE-19 cells and primary human RPE cells were co-transfected with two plasmids encoding the transposase and the transposon carrying a transcription cassette. We demonstrated efficient expression and secretion of BDNF in both RPE cell types, which were further increased in ARPE-19 cell cultures exposed to hydrogen peroxide. BDNF-transfected cells exhibited lower apoptosis rates and stimulated neurite outgrowth in human SH-SY5Y cells. This study is an important step in the development of a cell-based gene therapy that could be applied as an advanced therapy medicinal product to treat dry AMD or other degenerative retinal diseases.
越来越多的患者患有多种因素导致的神经退行性疾病，例如年龄相关性黄斑变性（AMD）。然而，其病理机制仍不清楚，这使得有效治疗方法的开发变得复杂。为了改善多种因素疾病的治疗效果，可以采用基于细胞的基因疗法来增加治疗因子的表达。迄今为止，尚无针对干性 AMD（包括晚期地图样萎缩）的获批疗法。我们提出了一种干性 AMD 的治疗选择，即通过质粒转座子系统的电穿孔将脑源性神经营养因子（BDNF）基因转入视网膜色素上皮（RPE）细胞。ARPE-19 细胞和原代人 RPE 细胞共转染两种质粒，分别编码转座酶和携带 BDNF 转录盒的转座子。我们证明了两种 RPE 细胞类型中 BDNF 的高效表达和分泌，并且在暴露于过氧化氢的 ARPE-19 细胞培养物中进一步增加。BDNF 转染的细胞凋亡率较低，并刺激人 SH-SY5Y 细胞的神经突生长。这项研究是开发基于细胞的 BDNF 基因疗法的重要一步，该疗法可作为先进的治疗药物产品用于治疗干性 AMD 或其他退行性视网膜疾病。
