title: PEGylated cationic nanoemulsions can efficiently bind and transfect pIDUA in a mucopolysaccharidosis type I murine model.
abstract: Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-L-iduronidase deficiency. This study proposed the use of cationic nanoemulsions as non-viral vectors for a plasmid (pIDUA) containing the gene that codes for alpha-L-iduronidase. Nanoemulsions composed of medium chain triglycerides (MCT)/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)/1,2-dioleoyl-sn-glycero-3-trimethylammonium propane (DOTAP)/1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG) were prepared by high pressure homogenization. Formulations were prepared by the adsorption or encapsulation of preformed pIDUA-DOTAP complexes into the oil core of nanoemulsions at different charge ratios. pIDUA complexed was protected from enzymatic degradation by DNase I. The physicochemical characteristics of complexes in protein-containing medium were mainly influenced by the presence of DSPE-PEG. Bragg reflections corresponding to a lamellar organization were identified for blank formulations by energy dispersive X-ray diffraction, which could not be detected after pIDUA complexation. The intravenous injection of these formulations in MPS I knockout mice led to a significant increase in IDUA activity (fluorescence assay) and expression (RT-qPCR) in different organs, especially the lungs and liver. These findings were more significant for formulations prepared at higher charge ratios (+4/-), suggesting a correlation between charge ratio and transfection efficiency. The present preclinical results demonstrated that these nanocomplexes represent a potential therapeutic option for the treatment of MPS I.
authors.affiliation: Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul (UFRGS), 90610-000 Porto Alegre, Brazil; Gene Therapy center, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA), 90035-903 Porto Alegre, Brazil.
authors.forename: Michelle
authors.initials: M
authors.lastname: Fraga
authors.affiliation: Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul (UFRGS), 90610-000 Porto Alegre, Brazil.
authors.forename: Fernanda
authors.initials: F
authors.lastname: Bruxel
authors.affiliation: Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul (UFRGS), 90610-000 Porto Alegre, Brazil.
authors.forename: Dirnete
authors.initials: D
authors.lastname: Diel
authors.affiliation: Gene Therapy center, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA), 90035-903 Porto Alegre, Brazil; Graduate Program in Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, Brazil.
authors.forename: Talita Giacomet
authors.initials: TG
authors.lastname: de Carvalho
authors.affiliation: Brazilian Synchrotron Light Laboratory, 13083-970 Campinas, São Paulo, Brazil.
authors.forename: Carlos Alberto
authors.initials: CA
authors.lastname: Perez
authors.affiliation: Physics Department, Universidade Federal de Minas Gerais, 31270-901 Belo Horizonte, Brazil.
authors.forename: Rogério
authors.initials: R
authors.lastname: Magalhães-Paniago
authors.affiliation: Physics Department, Universidade Federal de Minas Gerais, 31270-901 Belo Horizonte, Brazil.
authors.forename: Ângelo
authors.initials: Â
authors.lastname: Malachias
authors.affiliation: Pharmaceutical Products Department, Faculty of Pharmacy, Universidade Federal de Minas Gerais, 31270-901 Belo Horizonte, Brazil.
authors.forename: Mônica Cristina
authors.initials: MC
authors.lastname: Oliveira
authors.affiliation: Gene Therapy center, Experimental Research Center, Hospital de Clínicas de Porto Alegre (HCPA), 90035-903 Porto Alegre, Brazil; Graduate Program in Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, Brazil.
authors.forename: Ursula
authors.initials: U
authors.lastname: Matte
authors.affiliation: Graduate Program in Pharmaceutical Sciences, Universidade Federal do Rio Grande do Sul (UFRGS), 90610-000 Porto Alegre, Brazil. Electronic address: helder.teixeira@ufrgs.br.
authors.forename: Helder Ferreira
authors.initials: HF
authors.lastname: Teixeira
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D004655:Emulsions; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D015870:Gene Expression; D015316:Genetic Therapy; D006801:Humans; D007068:Iduronidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D008297:Male; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008059:Mucopolysaccharidosis I / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D049329:Nanostructures / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D010957:Plasmids; D011092:Polyethylene Glycols / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D013154:Spleen / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014280:Triglycerides / Q000737:chemistry
chemical_list: D004655:Emulsions; D005229:Fatty Acids, Monounsaturated; D010714:Phosphatidylethanolamines; D000644:Quaternary Ammonium Compounds; D014280:Triglycerides; C107570:polyethylene glycol-distearoylphosphatidylethanolamine; C094877:dioleoyl phosphatidylethanolamine; D011092:Polyethylene Glycols; D007068:Iduronidase; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane
keywords: Cationic nanoemulsions; DSPE-PEG; MPS I; Plasmid; pIDUA