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Forced expression of the Wilms tumor 1 (WT1) gene inhibits proliferation of human hematopoietic CD34+ progenitor cells

Leukemia volume 15, pages 1914–1922 (2001)Cite this article

Abstract

The Wilms tumor gene (WT1) encodes a zinc-finger containing transcription factor present in primitive hematopoietic progenitor cells. WT1 is also highly expressed in most cases of acute myeloid leukemia. Moreover, WT1 can interfere with induced differentiation of leukemic cell lines. These data suggest a function of WT1 in the maintenance of a primitive phenotype and a role in leukemogenesis by interfering with differentiation, prompting us to investigate its function in human hematopoietic progenitor cells. By retroviral transfer, human CD34+ cord blood progenitor cells were transduced with a vector encoding either of two splicing variants of WT1, with or without the KTS insert in the zinc-finger domain, linked to expression of green fluorescent protein (GFP) via an internal ribosomal entry site. When compared to cells transduced with vector containing GFP only, WT1 expressing cells showed strongly reduced colony formation in methylcellulose and inhibited proliferation in suspension culture, with no apparent reduction in viability. Cell cycle phase distribution was not affected by WT1 expression. No signs of impaired differentiation, as judged by the surface markers CD11b, CD14 and glycophorin were detected. In contrast to the results with human CD34+ progenitor cells, the proliferation of murine bone marrow cells was not significantly affected by WT1, consistent with previous data. We conclude that forced expression of WT1 in highly enriched human hematopoietic progenitor cells leads to strong anti-proliferative effects but is compatible with induced maturation of these cells.

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Acknowledgements

The authors wish to thank the staff at the Department of Obstetrics and Gynecology, Malmö University Hospital for invaluable help with the collection of cord blood samples. We thank Tor Olofsson for important and professional advice and the staff at ICH for help with the the flow cytometry analyses, Sverker Segrén for performing the flow cytometry cell sorting, and Thomas Relander for help with generation of retroviral vectors. The scientific advice and critical review of the manuscript by Inge Olsson is acknowledged. This work was supported by grants from the Swedish Cancer Society, the Swedish Medical Research Council (project No. 11546), the Swedish Childhood Cancer Foundation, the Bergquist Foundation, the Berta Kamprad Foundation, the Crafoord Foundation, the Georg Danielsson Foundation, the Gunnar Nilsson Cancer Foundation, the John Persson Foundation, the Thelma Zoega Foundation, the Greta and Johan Kock Foundation, the John and Augusta Persson Foundation, the Tobias Foundation, the Österlund Foundation, and from Funds of Lund University Hospital.

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  1. Dept of Hematology, Lund University, Lund, Sweden

    H Svedberg & U Gullberg

  2. Dept of Molecular Medicine and Gene Therapy, Lund University, Lund, Sweden

    J Richter

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Svedberg, H., Richter, J. & Gullberg, U. Forced expression of the Wilms tumor 1 (WT1) gene inhibits proliferation of human hematopoietic CD34+ progenitor cells. Leukemia 15, 1914–1922 (2001). https://doi.org/10.1038/sj.leu.2402303

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