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Expansion of genetically corrected neutrophils in chronic granulomatous disease mice by cotransferring a therapeutic gene and a selective amplifier gene

Gene Therapy volume 11, pages 1370–1377 (2004)Cite this article

Abstract

Hematopoietic stem cell gene therapy has not provided clinical success in disorders such as chronic granulomatous disease (CGD), where genetically corrected cells do not show a selective advantage in vivo. To facilitate selective expansion of transduced cells, we have developed a fusion receptor system that confers drug-induced proliferation. Here, a ‘selective amplifier gene (SAG)’ encodes a chimeric receptor (GcRER) that generates a mitotic signal in response to estrogen. We evaluated the in vivo efficacy of SAG-mediated cell expansion in a mouse disease model of X-linked CGD (X-CGD) that is deficient in the NADPH oxidase gp91phox subunit. Bone marrow cells from X-CGD mice were transduced with a bicistronic retrovirus encoding GcRER and gp91phox, and transplanted to lethally irradiated X-CGD recipients. Estrogen was administered to a cohort of the transplants, and neutrophil superoxide production was monitored. A significant increase in oxidase-positive cells was observed in the estrogen-treated mice, and repeated estrogen administration maintained the elevation of transduced cells for 20 weeks. In addition, oxidase-positive neutrophils were increased in the X-CGD transplants given the first estrogen even at 9 months post-transplantation. These results showed that the SAG system would enhance the therapeutic effects by boosting genetically modified, functionally corrected cells in vivo.

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Acknowledgements

We thank Dr MC Dinauer for X-CGD mice, and Dr M Nakamura for 7D5 monoclonal antibody. We are also grateful to Amgen for SCF, Ajinomoto for IL-6, Chugai Pharmaceuticals for Epo and G-CSF, and Takara Bio for RetroNectin. This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and the Ministry of Health, Labor and Welfare of Japan.

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Authors and Affiliations

  1. Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, Tochigi, Japan

    T Hara, A Kume, H Mizukami, T Okada & K Ozawa

  2. First Department of Internal Medicine, Gifu University School of Medicine, Gifu, Japan

    T Hara, H Tsurumi & H Moriwaki

  3. Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical School, Tochigi, Japan

    Y Hanazono

  4. DNAVEC Research Inc., Ibaraki, Japan

    Y Ueda & M Hasegawa

  5. Division of Hematology, Department of Medicine, Jichi Medical School, Tochigi, Japan

    K Ozawa

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  1. T Hara
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Hara, T., Kume, A., Hanazono, Y. et al. Expansion of genetically corrected neutrophils in chronic granulomatous disease mice by cotransferring a therapeutic gene and a selective amplifier gene. Gene Ther 11, 1370–1377 (2004). https://doi.org/10.1038/sj.gt.3302317

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