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CD34+ cells from dental pulp stem cells with a ZFN-mediated and homology-driven repair-mediated locus-specific knock-in of an artificial β-globin gene

Gene Therapy volume 24, pages 425–432 (2017)Cite this article

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Abstract

In humans, mutations in the β-globin gene (HBB) have two important clinical manifestations: β-thalassemia and sickle cell disease. The progress in genome editing and stem cell research may be relevant to the treatment of β-globin-related diseases. In this work, we employed zinc-finger nuclease (ZFN)-mediated gene integration of synthetic β-globin cDNA into HBB loci, thus correcting almost all β-globin mutations. The integration was achieved in both HEK 293 cells and isolated dental pulp stem cell (DPSCs). We also showed that DPSCs with an artificial gene knock-in were capable of generating stable six-cell clones and were expandable at least 108-fold; therefore, they may serve as a personalized stem cell factory. In addition, transfection with non-integrated pCAG-hOct4 and culturing in a conditioned medium converted the genome-edited DPSCs to CD34+ HSC-like cells. We believe that this approach may be useful for the treatment of β-globin-related diseases, especially the severe form of β-thalassemia.

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Acknowledgements

We thank Prof. Reiko Inagi (University of Tokyo, Tokyo) and Prof. Kanya Suphapeetiporn (Chulalongkorn Universty, Bangkok) for their helpful advice. We are grateful to Dr. Ryoji Sassa (Okasaki, Nagoya) for his generous support. This work was supported Lerdsin Foundation and SS Manufacturing.

Author contributions

SC: Conception and design, Collection and/or assembly of data, Data analysis and interpretation, Manuscript writing. OR: Conception and design, Collection and/or assembly of data, Data analysis and interpretation Financial support. WY: Provision of study material or patients. AS: Financial support, Administrative support. KM: Conception and design, Collection and/or assembly of data, Data analysis and interpretation, Manuscript writing, Administrative support Final approval of manuscript.

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Authors and Affiliations

  1. Department of Medicine, Molecular and Cell biology Unit, Lerdsin General Hospital, Bang-Rak Bangkok, Thailand

    S Chattong, W Yindeedej, A Setpakdee & K Manotham

  2. S.S. Manufacturing Co., Ltd 114/7 Moo2 Mahasawadi, Nonthaburi, Thailand

    S Chattong & O Ruangwattanasuk

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  1. S Chattong
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Correspondence to K Manotham.

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Competing interests

SC receives compensation as a consultant for SS Manufacturing Co., Ltd. OR is a share holder of SS Manufacturing Co., Ltd. The remaining authors declare no conflict of interest.

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Chattong, S., Ruangwattanasuk, O., Yindeedej, W. et al. CD34+ cells from dental pulp stem cells with a ZFN-mediated and homology-driven repair-mediated locus-specific knock-in of an artificial β-globin gene. Gene Ther 24, 425–432 (2017). https://doi.org/10.1038/gt.2017.42

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