title: A novel homologous model for gene therapy of dwarfism by non-viral transfer of the mouse growth hormone gene into immunocompetent dwarf mice.
abstract: The possibilities for non-viral GH gene therapy are studied in immunocompetent dwarf mice (lit/lit). As expression vector we used a plasmid previously employed in immunodeficient dwarf mice (pUBI-hGH-gDNA) by replacing the human GH gene with the genomic sequence of mouse-GH DNA (pUBI-mGH-gDNA). HEK-293 human cells transfected with pUBI-mGH-gDNA produced 3.0 µg mGH/10(6) cells/day compared to 3.7 µg hGH/10(6) cells/day for pUBIhGH- gDNA transfected cells. The weight of lit/lit mice treated with the same two plasmids (50 µg DNA/mouse) by electrotransfer into the quadriceps muscle was followed for 3 months. The weight increase up to 15 days for mGH, hGH and saline treated mice were 0.130, 0.112 and 0.027 g/mouse/day. Most sera from hGH-treated mice contained anti-hGH antibodies already on day 15, with the highest titers on day 45, while no significant anti-mGH antibodies were observed in mGH-treated mice. At the end of 3 months, the weight increase for mGH-treated mice was 34.3%, while the nose-to-tail and femur lengths increased 9.5% and 24.3%. Mouse-GH and hGH circulating levels were 4-5 ng/mL 15 days after treatment, versus control levels of ~0.7 ng GH/mL (P<0.001). In mGH-treated mice, mIGF-I determined on days 15, 45 and 94 were 1.5- to 3-fold higher than the control and 1.2- to 1.6-fold higher than hGH-treated mice. The described homologous model represents an important progress forming the basis for preclinical testing of non-viral gene therapy for GH deficiency.
authors.forename: Claudia R
authors.initials: CR
authors.lastname: Cecchi
authors.forename: Eliza
authors.initials: E
authors.lastname: Higuti
authors.forename: Nelio A J
authors.initials: NA
authors.lastname: Oliveira
authors.forename: Eliana R
authors.initials: ER
authors.lastname: Lima
authors.forename: Maria
authors.initials: M
authors.lastname: Jakobsen
authors.forename: Frederick
authors.initials: F
authors.lastname: Dagnaes-Hansen
authors.forename: Hanne
authors.initials: H
authors.lastname: Gissel
authors.forename: Lars
authors.initials: L
authors.lastname: Aagaard
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Jensen
authors.forename: Alexander A L
authors.initials: AA
authors.lastname: Jorge
authors.forename: Paolo
authors.initials: P
authors.lastname: Bartolini
authors.affiliation: Biotechnology Center, IPENCNEN, Avenida Professor Lineu Prestes 2242, Cidade Universitaria, 05508- 000, Sao Paulo, SP, Brazil. cnperoni@ipen.br.
authors.forename: Cibele N
authors.initials: CN
authors.lastname: Peroni
mesh_terms: D000818:Animals; D001835:Body Weight; D004195:Disease Models, Animal; D004392:Dwarfism / Q000235:genetics* / Q000473:pathology / Q000628:therapy; D015316:Genetic Therapy; D013006:Growth Hormone / Q000235:genetics* / Q000627:therapeutic use; D057809:HEK293 Cells; D006801:Humans; D007121:Immunocompetence; D051379:Mice; D015430:Weight Gain / Q000235:genetics
chemical_list: D013006:Growth Hormone