Optimization of Mouse Growth Hormone Plasmid DNA Electrotransfer into Tibialis Cranialis Muscle of "Little" Mice

Autor(es)

Lima, Eliana Rosa ; Cecchi, Claudia Regina ; Higuti, Eliza ; Pacheco de Jesus, Gustavo Protasio ; Gomes, Alissandra Moura ; Zacarias, Enio Aparecido ; Bartolini, Paolo ; Peroni, Cibele Nunes

• Tipo de documento: Artigo Científico
• Fonte: Molecules; v. 25, n. 21, p. 9-pg., 2020-11-01.

Resumo

Previous non-viral gene therapy was directed towards two animal models of dwarfism: Immunodeficient (lit/scid) and immunocompetent (lit/lit) dwarf mice. The former, based on hGH DNA administration into muscle, performed better, while the latter, a homologous model based on mGH DNA, was less efficient, though recommended as useful for pre-clinical assays. We have now improved the growth parameters aiming at a complete recovery of the lit/lit phenotype. Electrotransfer was based on three pulses of 375 V/cm of 25 ms each, after mGH-DNA administration into two sites of each non-exposed tibialis cranialis muscle. A 36-day bioassay, performed using 60-day old lit/lit mice, provided the highest GH circulatory levels we have ever obtained for GH non-viral gene therapy: 14.7 +/- 3.7 ng mGH/mL. These levels, at the end of the experiment, were 8.5 +/- 2.3 ng/mL, i.e., significantly higher than those of the positive control (4.5 +/- 1.5 ng/mL). The catch-up growth reached 40.9% for body weight, 38.2% for body length and 82.6%-76.9% for femur length. The catch-up in terms of the mIGF-1 levels remained low, increasing from the previous value of 5.9% to the actual 8.5%. Although a complete phenotypic recovery was not obtained, it should be possible starting with much younger animals and/or increasing the number of injection sites. (AU)

• 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: 17/15503-9 - Terapia gênica por eletrotransferência do gene do hormônio de crescimento de camundongo em modelo animal de osteogênese imperfeita
• Beneficiário: Cibele Nunes Peroni
• Modalidade de apoio: Auxílio à Pesquisa - Regular