Molecular and Functional Characterization of BDNF-Overexpressing Human Retinal Pigment Epithelial Cells Established by Sleeping Beauty Transposon-Mediated Gene Transfer

• Published in: International journal of molecular sciences Vol. 23; no. 21; p. 12982
• Main Authors: Mattern, Larissa, Otten, Katrin, Miskey, Csaba, Fuest, Matthias, Izsvák, Zsuzsanna, Ivics, Zoltán, Walter, Peter, Thumann, Gabriele, Johnen, Sandra
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.11.2022; MDPI
• Subjects: Apoptosis; Atrophy; Axonogenesis; BDNF; Brain-derived neurotrophic factor; cell-based additive gene therapy; Clinical trials; Diabetic retinopathy; Electroporation; Epithelium; Experiments; Gene expression; Gene therapy; Genetic engineering; Glaucoma; Hydrogen peroxide; Kinases; Macular degeneration; Neurodegeneration; neurodegenerative retinal diseases; non-viral transfection; Plasmids; Proteins; Ratios; Retina; Retinal pigment epithelium; RPE cells; Sleeping Beauty transposon system; Stem cells; Transposase; Transposons
• ISSN: 1422-0067; 1661-6596; 1422-0067
• 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 (BDNF) gene into retinal pigment epithelial (RPE) cells by electroporation using the plasmid-based Sleeping Beauty (SB) transposon system. ARPE-19 cells and primary human RPE cells were co-transfected with two plasmids encoding the SB100X transposase and the transposon carrying a BDNF 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 BDNF gene therapy that could be applied as an advanced therapy medicinal product to treat dry AMD or other degenerative retinal diseases.