Abstract
Preclinical studies in several animal models as well as clinical trials have shown a reduction in tumor growth following immunotherapy with interleukin-12 (IL-12). This cytokine is appropriate to test in therapeutic clinical trials to treat hepatocarcinoma (HC), a pathology often associated with hepatitis B or C-induced cirrhosis. The local delivery into the liver would be achieved through ex vivo gene transfer using retroviral (rv) vectors in autologous fibroblast carriers. In support of this clinical trial, a rv vector has been constructed to express coordinately both chains p35 and p40 of human IL-12. Here, we have tested good manufacturing practices (GMP) clinical lots of viral vectors derived from the transfected packaging cell line, PG13rvIL-12. We have also devised methods to facilitate the isolation of fibroblasts from freshly harvested skin specimens, enhance their outgrowth in large-scale cultures and assay IL-12 production following transduction, without any selection and irradiation. Twenty-four human skin specimens were processed to obtain fibroblast suspensions that were typically maintained for up to 8 or 12 passages. The mean ±s.d. overall time for obtaining the required number of transduced cells for the highest IL-12 need was 40 days. The procedure, in accordance with the French medical agency for gene therapy clinical trials, is now ready to begin a clinical trial.
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Acknowledgements
We are grateful to Genethon, (Patricia Noguiey's team) which constructed the pMFG-hIL-12 plasmid and synthesized the clinical-grade retroviral vectors. We thank Dr J. Bonnet who performed the γ-irradiation of the cells. We are also grateful to Emily Witty (AngloScribe) for the careful reading and correction of this paper. We also thank Pr JP Vinel and Pr L Buscail for their critical input. This study was supported by La ligue contre le cancer, The Region Midi Pyrenees, the GRICR (Groupe de Recherche Institut Claudius Regaud) and hospitals of Toulouse III (PHRC; France).
Grant support: La ligue contre le cancer, the Region Midi Pyrenees, the GRICR (Groupe de Recherche Institut Claudius Regaud) and hospitals of Toulouse III (PHRC; France).
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Lortal, B., Gross, F., Peron, J. et al. Preclinical study of an ex vivo gene therapy protocol for hepatocarcinoma. Cancer Gene Ther 16, 329–337 (2009). https://doi.org/10.1038/cgt.2008.88
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DOI: https://doi.org/10.1038/cgt.2008.88
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