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Persisting multilineage transgene expression in the clonal progeny of a hematopoietic stem cell

Leukemia volume 16, pages 1655–1663 (2002)Cite this article

Abstract

Many applications of hematopoietic gene therapy require selection for clones with active transgene expression. However, it was unclear whether the clonal progeny of a retrovirally transduced hematopoietic stem cell would be capable of maintaining transgene expression through serial repopulation and multilineage differentiation. Such investigations require simultaneous analyses of clonality, multilineage activity and transgene copy numbers. Using a mouse model, the present study demonstrates that a single hematopoietic stem cell expressing a marker gene from one or two insertions of a simple retroviral vector actively maintains multilineage transgene expression in the vast majority (80–99%) of bone marrow and peripheral blood cells. Gene expression persisted through serial transplantations for at least 97 weeks post gene transfer and was observed in the lymphoid (B, T and NK cells), myeloid (CD11b+, Gr-1+), erythroid (Ter119+, mature red blood cells) and megakaryocytic (as indicated by platelets) progeny. Therefore, a single immunoselection for hematopoietic stem cells expressing the transgene in vivo was sufficient to establish a completely chimeric hematopoiesis. These observations imply that the retroviral vectors used in this study contain cis-elements that mediate expression through massive clonal expansion and multilineage differentiation, provided the insertion occurred in genetic loci permissive for expression in hematopoietic stem cells.

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Acknowledgements

This work was supported by the Deutsche Krebshilfe (10–1456-Ba2, animal models), the BMBF (development of selection systems BMBF 01KV981, and BMBF 0312173 via CellTec), the VolkswagenStiftung, and the DAAD (personal grant to ZL). ZL also received support of the National Science Foundation of China (39900064), Guangdong Provincial Natural Science Foundation of China (990105) and the Chinese Ministry of Education (grants 2000/11 and 2000/65). The Heinrich Pette Institute is supported financially by the Bundesministerium für Gesundheit and by the Freie und Hansestadt Hamburg. The authors wish to thank Gökhan Arman-Kalcek, Dung Ung, Cordula Grüttner, Susanne Feldhaus and Maike Ziesnitz for expert technical assistance.

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Authors and Affiliations

  1. Department of Hematology and Oncology, Experimental Cell Therapy, Hannover Medical School, Hannover, Germany

    Z Li & C Baum

  2. Dept of Cell and Virus Genetics, Heinrich-Pette-Institute, Hamburg, Germany

    Z Li, B Schiedlmeier, O Frank, W Ostertag & C Baum

  3. Dept Hematology, Sun Yat-Sen Memorial Hospital, Guangzhou, China

    Z Li

  4. Bone Marrow Transplantation, University Hospital Eppendorf, Hamburg, Germany

    B Fehse & AR Zander

  5. Neuroanatomy, University Hospital Eppendorf, Hamburg, Germany

    J Düllmann

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Li, Z., Fehse, B., Schiedlmeier, B. et al. Persisting multilineage transgene expression in the clonal progeny of a hematopoietic stem cell. Leukemia 16, 1655–1663 (2002). https://doi.org/10.1038/sj.leu.2402619

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