An homologous model of gene therapy by in vivo administration of a plasmid containing the mouse growth hormone gene in immunocompetent dwarf mice

TY - CONF

T1 - An homologous model of gene therapy by in vivo administration of a plasmid containing the mouse growth hormone gene in immunocompetent dwarf mice

AU - Cecchi, Claudia R.

AU - Dagnæs-Hansen, Niels Frederik

AU - Aagaard, Lars

AU - Jakobsen, Maria Vad

AU - Gissel, Hanne

AU - Gothelf, Anita Birgitte

AU - Hansen, Birgit Holm

AU - Frystyk, Jan

AU - Higuti, Eliza

AU - Oliveira, Nelio A J

AU - Lima, Eliana R

AU - Bartolini, Paolo

AU - Jensen, Thomas G.

AU - Peroni, Cibele N

PY - 2012

Y1 - 2012

N2 - An alternative method for growth hormone deficiency (GHD )treatment has been developed using gene therapy associated to electroporation. Sustained circulatory levels of hGH and a highly significant weight increase were obtained in immunodeficient dwarf (lit/scid) mice. This new study aimed at setting up an homologous model of gene therapy based on electrotransfer of genomic mouse growth hormone (gmGH) in the muscle of im-munocompetent dwarf (lit/lit) mice. The plasmid pUC-UBI-gmGH was first transfected into HEK 293 cells and an in vitro expression up to 74.0 ng mGH/106 cells/day, compared to 6.0 ng mGH/106 cells/day for the negative control, was obtained. As apositive control for in vivo expression, this plasmid was administered as naked DNA via hydrodynamic injection into C57BL6 normal mice, and the weight increase of treated mice after 21days was 60.10%–11.02% compared to-2.05%–2.32% for thecontrol group (injected with Ringer’s solution). The plasmid was then administered in a protocol of gene-electrotransfer, applying eight 150 V/cm pulses of 20 ms, for a body weight gain assay into lit/lit mice. The weight increase after 15 days was 13.48%–3.53%for the gmGH-treated group versus 5.66%–2.51% for the control group (injected with saline) and 13.10%–7.54% for the group injected with the same plasmid encoding the ghGH gene, both groups also followed by electroporation. This previous results can be considered very promising as a pre-clinical study in a gene therapy protocol for patients suffering for GHD.

AB - An alternative method for growth hormone deficiency (GHD )treatment has been developed using gene therapy associated to electroporation. Sustained circulatory levels of hGH and a highly significant weight increase were obtained in immunodeficient dwarf (lit/scid) mice. This new study aimed at setting up an homologous model of gene therapy based on electrotransfer of genomic mouse growth hormone (gmGH) in the muscle of im-munocompetent dwarf (lit/lit) mice. The plasmid pUC-UBI-gmGH was first transfected into HEK 293 cells and an in vitro expression up to 74.0 ng mGH/106 cells/day, compared to 6.0 ng mGH/106 cells/day for the negative control, was obtained. As apositive control for in vivo expression, this plasmid was administered as naked DNA via hydrodynamic injection into C57BL6 normal mice, and the weight increase of treated mice after 21days was 60.10%–11.02% compared to-2.05%–2.32% for thecontrol group (injected with Ringer’s solution). The plasmid was then administered in a protocol of gene-electrotransfer, applying eight 150 V/cm pulses of 20 ms, for a body weight gain assay into lit/lit mice. The weight increase after 15 days was 13.48%–3.53%for the gmGH-treated group versus 5.66%–2.51% for the control group (injected with saline) and 13.10%–7.54% for the group injected with the same plasmid encoding the ghGH gene, both groups also followed by electroporation. This previous results can be considered very promising as a pre-clinical study in a gene therapy protocol for patients suffering for GHD.

U2 - 10.1089/hum.2012.2519

DO - 10.1089/hum.2012.2519

M3 - Poster

T2 - Annual Congress of the European Society of Gene and Cell Therapy

Y2 - 26 October 2012 through 29 October 2012

ER -