Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model

Michelle Fraga,1,2 Talita Giacomet de Carvalho,2,3 Juliana Bidone,1 Roselena Silvestri Schuh,1,2 Ursula Matte,2,3 Helder Ferreira Teixeira1 1Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Cl&ia...

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Autores principales:	Fraga M, de Carvalho TG, Bidone J, Schuh RS, Matte U, Teixeira HF
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2017
Materias:	Cationic nanoemulsions DSPE-PEG MPS I plasmid pIDUA. Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/56d3c5511bd34b378d8f2e6e8732a085
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spelling	oai:doaj.org-article:56d3c5511bd34b378d8f2e6e8732a0852021-12-02T03:54:16ZFactors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model1178-2013https://doaj.org/article/56d3c5511bd34b378d8f2e6e8732a0852017-03-01T00:00:00Zhttps://www.dovepress.com/factors-influencing-transfection-efficiency-of-piduananoemulsion-compl-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Michelle Fraga,1,2 Talita Giacomet de Carvalho,2,3 Juliana Bidone,1 Roselena Silvestri Schuh,1,2 Ursula Matte,2,3 Helder Ferreira Teixeira1 1Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre, 3Genetics and Molecular Biology Graduate Program, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil Abstract: Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-L-iduronidase (IDUA) deficiency. This study used IDUA knockout 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 in IDUA expression 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. Keywords: cationic nanoemulsions, DSPE-PEG, MPS I, plasmid, pIDUAFraga Mde Carvalho TGBidone JSchuh RSMatte UTeixeira HFDove Medical PressarticleCationic nanoemulsionsDSPE-PEGMPS IplasmidpIDUA.Medicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 12, Pp 2061-2067 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cationic nanoemulsions DSPE-PEG MPS I plasmid pIDUA. Medicine (General) R5-920
spellingShingle	Cationic nanoemulsions DSPE-PEG MPS I plasmid pIDUA. Medicine (General) R5-920 Fraga M de Carvalho TG Bidone J Schuh RS Matte U Teixeira HF Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
description	Michelle Fraga,1,2 Talita Giacomet de Carvalho,2,3 Juliana Bidone,1 Roselena Silvestri Schuh,1,2 Ursula Matte,2,3 Helder Ferreira Teixeira1 1Pharmaceutical Sciences Graduate Program, Universidade Federal do Rio Grande do Sul, 2Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre, 3Genetics and Molecular Biology Graduate Program, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil Abstract: Mucopolysaccharidosis type I (MPS I) is an autosomal disease caused by alpha-L-iduronidase (IDUA) deficiency. This study used IDUA knockout 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 in IDUA expression 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. Keywords: cationic nanoemulsions, DSPE-PEG, MPS I, plasmid, pIDUA
format	article
author	Fraga M de Carvalho TG Bidone J Schuh RS Matte U Teixeira HF
author_facet	Fraga M de Carvalho TG Bidone J Schuh RS Matte U Teixeira HF
author_sort	Fraga M
title	Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_short	Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_full	Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_fullStr	Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_full_unstemmed	Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model
title_sort	factors influencing transfection efficiency of pidua/nanoemulsion complexes in a mucopolysaccharidosis type i murine model
publisher	Dove Medical Press
publishDate	2017
url	https://doaj.org/article/56d3c5511bd34b378d8f2e6e8732a085
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Factors influencing transfection efficiency of pIDUA/nanoemulsion complexes in a mucopolysaccharidosis type I murine model

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