Efficient Non-Invasive Plasmid-DNA Administration into Tibialis Cranialis Muscle of "Little" Mice

Autor(es)

Cecchi, C. R. ; Higuti, E. ; Lima, E. R. ; Vieira, D. P. ; Squair, P. L. ; Peroni, C. N. ; Bartolini, P.

• Tipo de documento: Artigo Científico
• Fonte: CURRENT MOLECULAR MEDICINE; v. 17, n. 3, p. 6-pg., 2017-01-01.

Resumo

Background: An alternative treatment for growth hormone deficiency based on hGH-DNA administration, followed by electro gene transfer, was investigated by injecting the plasmid into surgically exposed or non-exposed quadriceps or tibialis muscle of immunodeficient "little" mice. Methods: An optimization of electrotransfer conditions via a new combination of high/low voltage pulses is presented. After 3 days, serum hGH was determined and in a 28-day assay, the relative growth parameters were compared. Results: Both groups exhibited similar results: 5.0 +/- 2.2 (SD) and 3.5 +/- 0.9 ng hGH/ml (P>0.05; n=7) for the exposed quadriceps and non-exposed tibialis treatments, respectively. The final body weight increases were 16.1% for the quadriceps and 18.9% for the tibialis group. The tail and nose-to-tail length increases were 4.5% and 7.1% for the quadriceps and 4.8 and 4.6% for the tibialis group. The right and left femur length increases, obtained from radiographic measurements, were 16.9% and 12.7% for the quadriceps and 19.4% and 12.3% for the tibialis, respectively. A non-significant difference between exposed quadriceps and non-exposed tibialis treatments (P=0.48) was confirmed via a completely integrated statistical analysis. Circulating mIGF-1 levels were 126 +/- 47, 106 +/- 93 (P>0.05) and 38 +/- 15 ng/ml for the quadriceps, tibialis and saline treatments, respectively. Conclusion: These results show that hGH-DNA administration into non-exposed tibialis muscle followed by the new HV/LV electrotransfer protocol was an equally efficient, less traumatic treatment, much more suitable for pre-clinical testing than administration into exposed quadriceps. (AU)

• 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: 14/19757-7 - Terapia gênica baseada em hormônio de crescimento: injeção de DNA plasmidial e implante de células microencapsuladas em camundongos anões
• Beneficiário: Cibele Nunes Peroni
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/07380-6 - Otimização de parâmetros de transferência in vivo do gene do hormônio de crescimento visando a correção fenotípica de camundongos anões
• Beneficiário: Eliana Rosa Lima Filha
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 14/04277-0 - 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 Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 14/18242-3 - Terapia gênica para a deficiência de hormônio de crescimento mediada pela eletrotransferência de minicírculos de DNA em camundongos anões
• Beneficiário: Eliza Higuti Sousa
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado