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

Hideto Matsui; Naoko Fujimoto; Noriko Sasakawa; Yasuhide Ohinata; Midori Shima; Shinya Yamanaka; Mitsuhiko Sugimoto; Akitsu Hotta

doi:10.1371/journal.pone.0104957

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

PLoS One. 2014 Aug 15;9(8):e104957. doi: 10.1371/journal.pone.0104957. eCollection 2014.

Authors

Hideto Matsui¹, Naoko Fujimoto², Noriko Sasakawa³, Yasuhide Ohinata⁴, Midori Shima⁵, Shinya Yamanaka⁶, Mitsuhiko Sugimoto¹, Akitsu Hotta⁷

Affiliations

¹ Department of Regulatory Medicine for Thrombosis, Nara Medical University, Kashihara, Nara, Japan.
² Department of Reprogramming Science, Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, Japan; iCeMS, Kyoto University, Kyoto, Japan.
³ Department of Reprogramming Science, Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, Japan.
⁴ 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.
⁵ Pediatrcs, Nara Medical University, Kashihara, Nara, Japan.
⁶ 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.
⁷ 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.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA Transposable Elements
DNA, Complementary / administration & dosage
DNA, Complementary / genetics
DNA, Complementary / therapeutic use*
Disease Models, Animal
Factor VIII / analysis
Factor VIII / genetics*
Gene Expression
Gene Transfer Techniques
Genetic Therapy / methods
Genetic Vectors / administration & dosage
Genetic Vectors / genetics
Genetic Vectors / therapeutic use*
HEK293 Cells
Hemophilia A / blood
Hemophilia A / genetics
Hemophilia A / therapy*
Humans
Mice

Substances

DNA Transposable Elements
DNA, Complementary
Factor VIII

Grants and funding

This work was supported by JSPS KAKENHI (No. 25293237 for HM, M. Shima, and AH, No.24591559 for HM, No. 22890088 for AH), research grants from the Bayer Hemophilia Awards Program (http://www.bayer-hemophilia-awards.com/) (HM), the Mochida Memorial Foundation for Medical and Pharmaceutical Research (http://www.mochida.co.jp/zaidan/) (HM), the Uehara Memorial Foundation (http://www.ueharazaidan.or.jp/) (AH), and the Baxter Coagulation Research Fund (http://baxter.co.jp/medical/grant/bhf/index.html) (AH). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.