Delivery of full-length factor VIII using a piggyBac transposon vector to correct a mouse model of hemophilia A

• Published in: PloS one 2014-08, Vol.9 (8), p.e104957-e104957
• Main Authors: Matsui, Hideto, Fujimoto, Naoko, Sasakawa, Noriko, Ohinata, Yasuhide, Shima, Midori, Yamanaka, Shinya, Sugimoto, Mitsuhiko, Hotta, Akitsu
• Format: Article
• Language: English
• Subjects: Adenoviruses; Animals; Antibodies; Biocompatibility; Biology and Life Sciences; Blood coagulation; Cell culture; Coagulation; Coagulation factors; Culture media; Cytotoxicity; Deoxyribonucleic acid; Disease; Disease Models, Animal; DNA; DNA Transposable Elements; DNA, Complementary - administration & dosage; DNA, Complementary - genetics; DNA, Complementary - therapeutic use; Expression vectors; Factor VIII - analysis; Factor VIII - genetics; Factor VIII deficiency; Gene Expression; Gene therapy; Gene transfer; Gene Transfer Techniques; Genetic Therapy - methods; Genetic Vectors - administration & dosage; Genetic Vectors - genetics; Genetic Vectors - therapeutic use; Genomes; HEK293 Cells; Hemophilia; Hemophilia A - blood; Hemophilia A - genetics; Hemophilia A - therapy; Humans; Immunology; Media (culture); Medicine and health sciences; Mice; Science; Thrombosis; Toxicity; Transgenes; Transposons; Trichoplusia ni; Vectors (Biology); Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0104957

Viral vectors have been used for hemophilia A gene therapy. However, due to its large size, full-length Factor VIII (FVIII) cDNA has not been successfully delivered using conventional viral vectors. Moreover, viral vectors may pose safety risks, e.g., adverse immunological reactions or virus-mediated cytotoxicity. Here, we took advantages of the non-viral vector gene delivery system based on piggyBac DNA transposon to transfer the full-length FVIII cDNA, for the purpose of treating hemophilia A. We tested the efficiency of this new vector system in human 293T cells and iPS cells, and confirmed the expression of the full-length FVIII in culture media using activity-sensitive coagulation assays. Hydrodynamic injection of the piggyBac vectors into hemophilia A mice temporally treated with an immunosuppressant resulted in stable production of circulating FVIII for over 300 days without development of anti-FVIII antibodies. Furthermore, tail-clip assay revealed significant improvement of blood coagulation time in the treated mice. piggyBac transposon vectors can facilitate the long-term expression of therapeutic transgenes in vitro and in vivo. This novel gene transfer strategy should provide safe and efficient delivery of FVIII.