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Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy

Gene Therapy volume 6, pages 1038–1044 (1999)Cite this article

Abstract

We have proposed a novel concept, ie selective expansion of transduced cells, to overcome the low efficiency of gene transfer into hematopoietic stem cells. Previously, a fusion gene encoding a chimeric receptor (ΔGCRER) between the mouse granulocyte colony-stimulating factor receptor (G-CSFR) and the hormone-binding domain of rat estrogen receptor was constructed as a ‘selective amplifier gene’. Although the chimeric gene conferred estrogen-inducible proliferation on the transduced Ba/F3 cells, it also mediated differentiation of the retrovirally transduced 32D cells upon estrogen treatment. Since only a growth signal is required for our purpose, we further modified the ΔGCRER gene to attenuate its differentiation signal. Based on the observation that tyrosine-703 in wild-type G-CSFR plays a pivotal role in transmitting the differentiation signal, phenylalanine was substituted for this residue in ΔGCRER. When the resultant selective amplifier gene (ΔY703F-GCRER gene) was expressed in 32D cells, sustained growth was supported by estrogen, while differentiation was suppressed. These cells ceased to grow upon estrogen withdrawal and differentiated with G-CSF treatment. The present findings suggested that ΔY703F-GCRER may have desirable properties as a selective amplifier for hematopoietic stem cell expansion and gene therapy.

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Acknowledgements

We thank Dr JM Cunningham, Dr T Suda and Chugai Pharmaceuticals for research materials, and Dr A Okano for IL-3 titration. This work was supported in part by grants from the Ministry of Health and Welfare of Japan and Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.

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Authors and Affiliations

  1. Division of Genetic Therapeutics, Center for Molecular Medicine, Saitama, Japan

    K M Matsuda, A Kume, Y Ueda, M Urabe & K Ozawa

  2. CREST, Japan Science and Technology Corporation, Saitama, Japan

    K M Matsuda, A Kume, Y Ueda, M Urabe & K Ozawa

  3. DNAVEC Research Inc, Ibaraki, Japan

    Y Ueda & M Hasegawa

  4. Department of Hematology, Jichi Medical School, Saitama, Japan

    K Ozawa

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  1. K M Matsuda
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  2. A Kume
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  3. Y Ueda
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  4. M Urabe
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Matsuda, K., Kume, A., Ueda, Y. et al. Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy. Gene Ther 6, 1038–1044 (1999). https://doi.org/10.1038/sj.gt.3300906

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