ROSANGELA DO ROCIO ARKATEN

ROSANGELA DO ROCIO ARKATEN

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Approaches to maximizing the periplasmic expression in Escherichia coli of the human growth hormone gene from a vector containing the lambda Psub(L)
2002 - SOARES, C.R.J.; OLIVEIRA, J.E.; SOUSA, J.M.; ARKATEN, R.R.; BARTOLINI, P.
Retrovirus-mediated transfer of growth hormone gene into primary human keratinocytes produces measurable circulatory levels of the hormone in immunodeficient-dwarf mice
2001 - BELLINI, M.H.; ROSAURO, C.W.; PERONI, C.N.; ARKATEN, R.R.; BARTOLINI, P.
Enrichment of human keratinocyte stem cells and inection of naked DNA in the lit/scid mouse: studies directed to the improvement of GH gene therapy
2006 - CECCHI, C.R.; PERONI, C.N.; DAMIANI, R.; DANTAS, E.O.; ARKATEN, R.R.; BARTOLINI, P.
High-level secretion of growth hormone by retrovirally transduced primary human keratinocytes: prospects for an animal model of cutaneous gene therapy
2005 - PERONI, C.N.; CECCHI, C.R.; DAMIANI, R.; SOARES, C.R.J.; RIBELA, M.T.C.P.; ARKATEN, R.R.; BARTOLINI, P.
Hollow fiber bioreactors for hormone production in CHO cells
2000 - SOARES, C.R.J.; PERONI, C.N.; MORGANTI, L.; AFFONSO, R.; ARKATEN, R.R.; SOUZA, J.M.; BARTOLINI, P.
High-level secretion of growth hormone by retrovirally transduced primary human keratinocytes
2006 - PERONI, CIBELE N.; CECCHI, CLAUDIA R.; DAMIANI, RENATA; SOARES, CARLOS R.J.; RIBELA, MARIA T.C.P.; ARKATEN, ROSANGELA R.; BARTOLINI, PAOLO
A gene therapy clinical trial for treatment of growth hormone (GH) deficiency has not been reached yet, but several strategies using different gene transfer methodologies and animal models have been developed and showed successful results. We have set up an ex vivo gene therapy protocol using primary human keratinocytes transduced with an efficient retroviral vector (LXSN) encoding the human (hGH) or mouse GH (mGH) genes. These stably modified cells presented high in vitro expression levels of hGH (7 μg/106 cells/d) and mGH (11 μg/106 cells/d) after selection with geneticin. When the hGH-secreting keratinocytes were grafted onto immunodeficient dwarf mice (lit/scid), hGH levels in the circulation were about 0.2–0.3 ng/mL during a 12-d assay and these animals presented a significant body weight increase (p<0.01) compared to the control. Substitution of conventional grafting methodologies with organotypic raft cultures revealed a peak value of up to 20 ng mGH/mL in the circulation of grafted lit/scid mice at 1 h postimplantation, followed by a rapid decline to baseline (≈2 ng/mL) within 24 h. One week after grafting, however, the cultured excised implants still presented approx 45% of their original in vitro secretion efficiency. Further studies are being carrier out to identify the main factor(s) that still constitute one of the major impediments to the success of this promising model of cutaneous gene therapy.