title: Antiangiogenic and Neurogenic Activities of Sleeping Beauty-Mediated PEDF-Transfected RPE Cells In Vitro and In Vivo.
abstract: 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.
authors.affiliation: Department of Ophthalmology, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Johnen
authors.affiliation: Department of Ophthalmology, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Yassin
authors.initials: Y
authors.lastname: Djalali-Talab
authors.affiliation: Department of Ophthalmology, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Olga
authors.initials: O
authors.lastname: Kazanskaya
authors.affiliation: Department of Ophthalmology, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Theresa
authors.initials: T
authors.lastname: Möller
authors.affiliation: Department of Ophthalmology, University Hospitals of Geneva, Street Alcide-Jentzer 22, 1211 Geneva 14, Switzerland.
authors.forename: Nina
authors.initials: N
authors.lastname: Harmening
authors.affiliation: Department of Ophthalmology, University Hospitals of Geneva, Street Alcide-Jentzer 22, 1211 Geneva 14, Switzerland.
authors.forename: Martina
authors.initials: M
authors.lastname: Kropp
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Robert-Rössle-Straße 10, 13125 Berlin, Germany.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.affiliation: Department of Ophthalmology, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Walter
authors.affiliation: 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.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Thumann
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D020256:Choroidal Neovascularization / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D004251:DNA Transposable Elements / Q000235:genetics; D005136:Eye Proteins / Q000008:administration & dosage / Q000235:genetics*; D045888:Ganglion Cysts / Q000235:genetics / Q000473:pathology; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism / Q000637:transplantation*; D006801:Humans; D008268:Macular Degeneration / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D009414:Nerve Growth Factors / Q000008:administration & dosage / Q000235:genetics*; D016501:Neurites / Q000378:metabolism / Q000473:pathology; D010786:Photoreceptor Cells / Q000378:metabolism / Q000473:pathology; D051381:Rats; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics*; D055213:Retinal Pigment Epithelium / Q000378:metabolism / Q000473:pathology; D015843:Serpins / Q000008:administration & dosage / Q000235:genetics*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D004251:DNA Transposable Elements; D005136:Eye Proteins; D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D015843:Serpins; D042461:Vascular Endothelial Growth Factor A; D000099174:Pigment Epithelium-Derived Factor