title: Nonviral in vivo gene transfer in the mucopolysaccharidosis I murine model.
abstract: Mucopolysaccharidosis I (MPS I) is a lysosomal disorder characterized by a deficiency of the enzyme alpha-L: -iduronidase (IDUA), which is responsible for the degradation of glycosaminoglycans (GAGs). This deficiency leads to the accumulation of dermatan and heparan sulphate in lysosomes. Presently available treatments include bone marrow transplantation and enzyme replacement therapies, both of which are limited in their effects. In this work, knockout (KO) MPS I mice were treated with a nonviral vector containing the human IDUA cDNA. KO mice were transfected by hydrodynamic injection of pRIDUA in the caudal vein (i.v., n = 3) or by intraperitoneal injection of pRIDUA/Superfect complexes (i.p., n = 3). GAG concentration and IDUA activity were analysed in the kidneys, spleen, lungs, brain and liver. The expression of IDUA in the organs of i.v.- and i.p.-treated mice was also analysed by real-time reverse-transcription (RT) PCR and compared by relative quantification. The concentration of GAGs in the organs differed between KO and wild-type mice. In the spleen and liver, GAG levels were lower in i.v.- and i.p.-treated KO mice than in control nontreated animals. Real-time RT-PCR showed that the transgene is expressed in all the analysed organs of i.p.- and i.v.-treated KO mice. Enzyme activity was similarly observed in all the organs analysed. Our data suggest that this kind of transfection may be a useful tool for studies of nonviral protocols for gene therapy of MPS.
authors.affiliation: Genetics Department, Universidade Federal do Rio Grande do Sul, Caixa Postal 15053, 91501-900, Porto Alegre, RS, Brasil.
authors.forename: M
authors.initials: M
authors.lastname: Camassola
authors.forename: L M
authors.initials: LM
authors.lastname: Braga
authors.forename: A
authors.initials: A
authors.lastname: Delgado-Cañedo
authors.forename: T P
authors.initials: TP
authors.lastname: Dalberto
authors.forename: U
authors.initials: U
authors.lastname: Matte
authors.forename: M
authors.initials: M
authors.lastname: Burin
authors.forename: R
authors.initials: R
authors.lastname: Giugliani
authors.forename: N B
authors.initials: NB
authors.lastname: Nardi
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation; D018076:DNA, Complementary / Q000378:metabolism; D003871:Dermatan Sulfate / Q000378:metabolism; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006025:Glycosaminoglycans / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008059:Mucopolysaccharidosis I / Q000235:genetics*; D010957:Plasmids / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D018076:DNA, Complementary; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D012333:RNA, Messenger