title: Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model.
abstract: Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-l-iduronidase (IDUA) deficiency. This study usedIDUAknockout mice as a model to evaluate whether parameters such as dose of plasmid and time of treatment could influence the transfection efficiency of complexes formed with PEGylated cationic nanoemulsions and plasmid (pIDUA), which contains the gene that encodes for IDUA. Formulations were composed of medium chain triglycerides, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(amino[polyethylene glycol]-2000), 1,2-dioleoyl-sn-glycero-3-trimethylammonium propane (DOTAP), glycerol, and water and were prepared by the adsorption or encapsulation of preformed pIDUA-DOTAP complexes by high-pressure homogenization. A progressive increase inIDUAexpression was observed with an increase in the dose and time of transfection for mice treated with both complexes (adsorbed and encapsulated), especially in the liver. Regardless of the complex administered, a significant increase in IDUA activity was detected in lungs and liver compared with nontreated MPS I when a dose of 60 μg was administered and IDUA activity was measured 7 days postadministration. Tissue sections of major organs showed no presence of cell necrosis, inflammatory infiltrate, or an increase in apoptosis. Furthermore, immunohistochemistry for CD68 showed no difference in the number of macrophage cells in treated and nontreated animals, indicating the absence of inflammatory reaction caused by the treatment. The data set obtained in this study allowed establishing that factors such as dose and time can influence transfection efficiency in different degrees and that these complexes did not lead to any lethal effect in the MPS I murine model used.
authors.affiliation: Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul; Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre.
authors.forename: Michelle
authors.initials: M
authors.lastname: Fraga
authors.affiliation: Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre; Genetics and Molecular Biology Graduate Program, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Talita Giacomet
authors.initials: TG
authors.lastname: de Carvalho
authors.affiliation: Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul.
authors.forename: Juliana
authors.initials: J
authors.lastname: Bidone
authors.affiliation: Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul; Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre.
authors.forename: Roselena Silvestri
authors.initials: RS
authors.lastname: Schuh
authors.affiliation: Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre; Genetics and Molecular Biology Graduate Program, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
authors.forename: Ursula
authors.initials: U
authors.lastname: Matte
authors.affiliation: Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul.
authors.forename: Helder Ferreira
authors.initials: HF
authors.lastname: Teixeira
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D004195:Disease Models, Animal; D004655:Emulsions / Q000737:chemistry*; D006801:Humans; D007068:Iduronidase / Q000378:metabolism*; D007150:Immunohistochemistry; D051379:Mice; D008059:Mucopolysaccharidosis I / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D000097382:CD68 Molecule
chemical_list: D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D004655:Emulsions; D007068:Iduronidase; C067980:CD68 antigen, human; D000097382:CD68 Molecule
keywords: DSPE-PEG; MPS I; cationic nanoemulsions; pIDUA; plasmid