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Expression of human coagulation factor VIII in adipocytes transduced with the simian immunodeficiency virus agmTYO1-based vector for hemophilia A gene therapy

Abstract

We demonstrate that transduction of adipocytes with a simian immunodeficiency virus agm TYO1 (SIVagm)-based lentiviral vector carrying the human coagulation factor VIII gene (SIVhFVIII) resulted in expression of the human FVIII transgene in vitro and in db/db mice in vivo. Cultured human adipocytes were transduced with the SIVagm vector carrying the GFP gene in a dose-dependent manner and transduction of adipocytes with SIVhFVIII resulted in efficient expression of human coagulation factor VIII (hFVIII; 320±39.8 ng/106 adipocytes/24 h) in vitro. Based upon successful transduction of adipocytes by SIV vectors carrying the lacZ gene in vivo in mice, the adipose tissue of db/db mice was transduced with SIVhFVIII. There was a transient appearance of human FVIII in mouse plasma (maximum 1.8 ng/ml) on day 11 after the injection. Transcripts of human FVIII transgene and human FVIII antigen also were detected in the adipose tissue by RT-PCR and immunofluorescence, respectively, on day 14. Emergence of anti-human FVIII antibodies 14 days after the injection of SIVhFVIII may explain the disappearance of human FVIII from the circulation. These results suggest that transduction of the adipocytes with vectors carrying the human FVIII gene may be potentially applicable for gene therapy of hemophilia A.

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Acknowledgements

We thank Dr JA van Mourik for the full-length FVIII cDNA and Dr Nakagaki (Chemo Sero Institute) for monoclonal antibodies to human FVIII. We also thank Dr DJ Stearns-Kurosawa (IdEst, Inc) for critical reading and editing of this manuscript, and Ms Fumino Muroi for technical assistance. This work is supported by Grants-in-aid for Scientific Research #12670687 to JM and #13671078 to SM from the Ministry of Education and Science, and by Health and Labour Sciences Research Grants for Research on HIV/AIDS to KO and to YS from the Ministry of Health, Labour and Welfare.

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Ogata, K., Mimuro, J., Kikuchi, J. et al. Expression of human coagulation factor VIII in adipocytes transduced with the simian immunodeficiency virus agmTYO1-based vector for hemophilia A gene therapy. Gene Ther 11, 253–259 (2004). https://doi.org/10.1038/sj.gt.3302174

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