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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References
Hoyer LW . Hemophilia A. N Engl J Med 1994; 330: 38–47.
Kay MA, High K . Gene therapy for the hemophilias. Proc Natl Acad Sci USA 1999; 96: 9973–9975.
Mohamed-Ali V, Pinkney JH, Coppack SW . Adipose tissue as an endocrine and paracrine organ. Int J Obes Relat Metab Disord 1998; 22: 1145–1158.
Kaufman RJ, Wasley LC, Dorner AJ . Synthesis, processing, and secretion of recombinant human factor VIII expressed in mammalian cells. J Biol Chem 1988: 6352–6362.
Levine JA, Eberhardt NL, Jensen MD, O'Brien T . Adenoviral-mediated gene transfer to human adipocytes in vitro, and human adipose tissue ex vivo and rabbit femoral adipose tissue in vivo. J Nutr Sci Vitaminol 1998; 44: 569–572.
Nagamatsu S et al. Adnovirus-medicated proproinsulin gene transfer into adipose tissues ameliorates hyperglycemia in obese diabetic KKAy mice. FEBS Lett 2001; 509: 106–110.
Miyoshi H et al. Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors. Science 1999; 283: 682–686.
Chen W et al. Lentiviral vector transduction of hematopoietic stem cells that mediate long-term reconstitution of lethally irradiated mice. Stem Cells 2000; 18: 352–359.
Woods NB et al. Lentiviral gene transfer into primary and secondary NOD/SCID repopulating cells. Blood 2000; 96: 3725–3733.
Naldini L et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 1996; 272: 263–267.
Honjo S et al. Experimental infection of African green monkeys and cynomolgus monkeys with a SIVAGM strain isolated from a healthy African green monkey. J Med Primatol 1990; 19: 9–20.
Roth DA, The factor FIII transkaryotic therapy study group et al. Implantation of non-viral ex vivo genetically modified autologous dermal fibroblaststhat express B-domain deleted human factor VIII in 12 severe hemophilia A study subjects. Blood 2002; 100 (Suppl): 116a (abstract #430).
Dwarki VJ et al. Gene therapy for hemophilia A: production of therapeutic levels of human factor VIII in vivo in mice. Proc Natl Acad Sci USA 1995; 14: 1023–1027.
Halvorsen YC et al. Thiazolinediones and glucocortidoids synergistically induce differentiation of human adipose tissue stromal cells: biochemical, cellular, and molecular analysis. Metabolism 2001; 50: 407–413.
Harp JB, Franklin D, Vanderpuije AA, Gimble JM . Differential expression of signal transducers and activators of transcription during human adipogenesis. Biochem Biophys Res Commun 2001; 281: 907–912.
Weiss GH, Rosen OM, Rubin CS . Regulation of fatty acid synthetase concentration and activity during adipocyte differentiation. J Biol Chem 1980; 255: 4751–4757.
Mertens K et al. Biological activity of recombinant factor VIII variants lacking the central B-domain and the heavy-chain sequence Lys713-Arg740: discordant in vitro and in vivo activity. Br J Haematol 1993; 85: 133–142.
Lind P et al. Novel forms of B-domain-deleted recombinant factor VIII molecules. Construction and biochemical characterization. Eur J Biochem 1995; 232: 19–27.
Nakajima T et al. Development of novel simian immunodeficiency virus vectors carrying a dual gene expression system. Hum Gene Ther 2000; 11: 1863–1874.
Naito M et al. Defective sorting to secretory vesicles in the trans Golgi network is partly responsible for protein C deficiency: molecular mechanisms of impaired secretion of abnormal protein C R169W, R352W, and G376D. Circ Res 2003; 92: 865–872.
Chua Jr S-C et al. Phenotypes of mouse diabetes and rat fatty due to mutations in the OB (leptin) receptor. Science 1996; 271: 994–996.
Yonemura H et al. Efficient production of recombinant human factor VIII by co-expression of the heavy and light chains. Protein Eng 1993; 6: 669–674.
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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DOI: https://doi.org/10.1038/sj.gt.3302174
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