A Novel Homologous Model for Gene Therapy of Dwarfism by Non-Viral Transfer of the Mouse Growth Hormone Gene into Immunocompetent Dwarf Mice

Autor(es):Mostrar menos -

Cecchi, Claudia R. ^[1] ; Higuti, Eliza ^[1] ; Oliveira, Nelio A. J. ^[1] ; Lima, Eliana R. ^[1] ; Jakobsen, Maria ^[2] ; Dagnaes-Hansen, Frederick ^[2] ; Gissel, Hanne ^[2] ; Aagaard, Lars ^[2] ; Jensen, Thomas G. ^[2] ; Jorge, Alexander A. L. ^[3] ; Bartolini, Paolo ^[1] ; Peroni, Cibele N. ^[1]

Afiliação do(s) autor(es)

^[1] Inst Pesquisas Energet & Nucl IPEN CNEN, Ctr Biotechnol, Sao Paulo - Brazil

^[2] Aarhus Univ, Dept Biomed, DK-8000 Aarhus C - Denmark

^[3] Univ Sao Paulo, Sch Med FMUSP, Dept Endocrinol, Genet Endocrinol Unit LIM25, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: CURRENT GENE THERAPY; v. 14, n. 1, p. 44-51, FEB 2014.
• Citações Web of Science: 6

Resumo

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 mu g mGH/10(6) cells/day compared to 3.7 mu g hGH/10(6) cells/day for pUBI-hGH-gDNA transfected cells. The weight of lit/lit mice treated with the same two plasmids (50 mu g 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 similar to 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. (AU)

• Processo FAPESP: 11/21708-6 - Completa correção fenotípica do nanismo mediante injeção de DNA plasmidial em modelo animal de deficiência isolada do hormônio de crescimento
• Beneficiário: Eliza Higuti Sousa
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 11/23365-9 - Eletrotransferência de um plasmídeo contendo o gene do hormônio de crescimento murino em camundongos anões para a obtenção de um modelo homólogo de terapia gênica in vivo
• Beneficiário: Eliana Rosa Lima Filha
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 13/03747-0 - Terapia gênica baseada em hormônio de crescimento: tratamento homólogo com DNA plasmidial e mediante implante de células microencapsuladas em camundongos anões imunocompetentes (lit/lit)
• Beneficiário: Claudia Regina Cecchi
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 10/51116-0 - Estudo dos efeitos endocrinos e paracrinos na administracao de dna plasmidial em modelos animais de terapia genica
• Beneficiário: Cibele Nunes Peroni
• Modalidade de apoio: Auxílio à Pesquisa - Regular