title: Delivery of full-length factor VIII using a piggyBac transposon vector to correct a mouse model of hemophilia A.
abstract: 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.
authors.affiliation: Department of Regulatory Medicine for Thrombosis, Nara Medical University, Kashihara, Nara, Japan.
authors.forename: Hideto
authors.initials: H
authors.lastname: Matsui
authors.affiliation: Department of Reprogramming Science, Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, Japan; iCeMS, Kyoto University, Kyoto, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Fujimoto
authors.affiliation: Department of Reprogramming Science, Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Sasakawa
authors.affiliation: Life Science Experimental Facility, Department of Biotechnology, Faculty of life and Environmental Sciences, University of Yamanashi, 4-4-37 Takeda, Kofu, Yamanashi, Japan; PRESTO, Japan Science and Technology Agency (JST), Saitama, Japan.
authors.forename: Yasuhide
authors.initials: Y
authors.lastname: Ohinata
authors.affiliation: Pediatrcs, Nara Medical University, Kashihara, Nara, Japan.
authors.forename: Midori
authors.initials: M
authors.lastname: Shima
authors.affiliation: Department of Reprogramming Science, Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, Japan; iCeMS, Kyoto University, Kyoto, Japan; Gladstone Institute of Cardiovascular Disease, San Francisco, California, United States of America.
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamanaka
authors.affiliation: Department of Regulatory Medicine for Thrombosis, Nara Medical University, Kashihara, Nara, Japan.
authors.forename: Mitsuhiko
authors.initials: M
authors.lastname: Sugimoto
authors.affiliation: Department of Reprogramming Science, Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, Japan; iCeMS, Kyoto University, Kyoto, Japan; PRESTO, Japan Science and Technology Agency (JST), Saitama, Japan.
authors.forename: Akitsu
authors.initials: A
authors.lastname: Hotta
mesh_terms: D000818:Animals; D004251:DNA Transposable Elements; D018076:DNA, Complementary / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use*; D004195:Disease Models, Animal; D005169:Factor VIII / Q000032:analysis / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use*; D057809:HEK293 Cells; D006467:Hemophilia A / Q000097:blood / Q000235:genetics / Q000628:therapy*; D006801:Humans; D051379:Mice
chemical_list: D004251:DNA Transposable Elements; D018076:DNA, Complementary; D005169:Factor VIII