title: Cationic polyene phospholipids as DNA carriers for ocular gene therapy.
abstract: Recent success in the treatment of congenital blindness demonstrates the potential of ocular gene therapy as a therapeutic approach. The eye is a good target due to its small size, minimal diffusion of therapeutic agent to the systemic circulation, and low immune and inflammatory responses. Currently, most approaches are based on viral vectors, but efforts continue towards the synthesis and evaluation of new nonviral carriers to improve nucleic acid delivery. Our objective is to evaluate the efficiency of novel cationic retinoic and carotenoic glycol phospholipids, designated C20-18, C20-20, and C30-20, to deliver DNA to human retinal pigmented epithelium (RPE) cells. Liposomes were produced by solvent evaporation of ethanolic mixtures of the polyene compounds and coformulated with 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) or cholesterol (Chol). Addition of DNA to the liposomes formed lipoplexes, which were characterized for binding, size, biocompatibility, and transgene efficiency. Lipoplex formulations of suitable size and biocompatibility were assayed for DNA delivery, both qualitatively and quantitatively, using RPE cells and a GFP-encoding plasmid. The retinoic lipoplex formulation with DOPE revealed a transfection efficiency comparable to the known lipid references 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]-cholesterol (DC-Chol) and 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (EPC) and GeneJuice. The results demonstrate that cationic polyene phospholipids have potential as DNA carriers for ocular gene therapy.
authors.affiliation: Center for Biomedicine and Structural Biology (CBME), Institute for Biotechnology and Bioengineering (IBB), University of Algarve, 8005-139 Faro, Portugal.
authors.forename: Susana
authors.initials: S
authors.lastname: Machado
authors.affiliation: Center for Biomedicine and Structural Biology (CBME), Institute for Biotechnology and Bioengineering (IBB), University of Algarve, 8005-139 Faro, Portugal ; Doctoral Program in Biomedical Sciences, Department of Biomedical Sciences and Medicine, University of Algarve, 8005-139 Faro, Portugal.
authors.forename: Sofia
authors.initials: S
authors.lastname: Calado
authors.affiliation: Center for Biomedicine and Structural Biology (CBME), Institute for Biotechnology and Bioengineering (IBB), University of Algarve, 8005-139 Faro, Portugal ; ProRegeM Doctoral Program, Department of Biomedical Sciences and Medicine, University of Algarve, 8005-139 Faro, Portugal.
authors.forename: Diogo
authors.initials: D
authors.lastname: Bitoque
authors.affiliation: Center for Biomedicine and Structural Biology (CBME), Institute for Biotechnology and Bioengineering (IBB), University of Algarve, 8005-139 Faro, Portugal ; Doctoral Program in Biomedical Sciences, Department of Biomedical Sciences and Medicine, University of Algarve, 8005-139 Faro, Portugal.
authors.forename: Ana Vanessa
authors.initials: AV
authors.lastname: Oliveira
authors.affiliation: Department of Chemistry, Norwegian University of Science and Technology, 7491 Trondheim, Norway.
authors.forename: Christer L
authors.initials: CL
authors.lastname: Øpstad
authors.affiliation: Premedical Unit, Weill Cornell Medical College in Qatar, P.O. Box 24144, Doha, Qatar.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Zeeshan
authors.affiliation: Department of Chemistry, Norwegian University of Science and Technology, 7491 Trondheim, Norway.
authors.forename: Hans-Richard
authors.initials: HR
authors.lastname: Sliwka
authors.affiliation: Department of Chemistry, Norwegian University of Science and Technology, 7491 Trondheim, Norway.
authors.forename: Vassilia
authors.initials: V
authors.lastname: Partali
authors.affiliation: Premedical Unit, Weill Cornell Medical College in Qatar, P.O. Box 24144, Doha, Qatar.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Pungente
authors.affiliation: Center for Biomedicine and Structural Biology (CBME), Institute for Biotechnology and Bioengineering (IBB), University of Algarve, 8005-139 Faro, Portugal ; Department of Biomedical Sciences and Medicine, University of Algarve, 8005-139 Faro, Portugal.
authors.forename: Gabriela A
authors.initials: GA
authors.lastname: Silva
mesh_terms: D002412:Cations / Q000378:metabolism*; D002460:Cell Line; D002784:Cholesterol / Q000031:analogs & derivatives / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000378:metabolism; D005123:Eye / Q000378:metabolism*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000378:metabolism*; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D011090:Polyenes / Q000378:metabolism*; D012168:Retinal Pigments / Q000378:metabolism; D014162:Transfection / Q000379:methods
chemical_list: D002412:Cations; D008081:Liposomes; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D011090:Polyenes; D012168:Retinal Pigments; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; C020888:1,2-dielaidoylphosphatidylethanolamine; D004247:DNA; D002784:Cholesterol