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Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity

Oncogene volume 21, pages 4778–4787 (2002)Cite this article

Abstract

STAT3 is a key downstream signaling intermediate of gp130, a receptor previously shown to activate hematopoietic stem cell (HSC) self-renewal divisions. These findings prompted us to investigate if the STAT3 pathway is important to HSC activity in vivo. Initial semi-quantitative RT–PCR analyses showed STAT3 to be expressed at slightly higher levels in primitive subsets of both human and murine adult bone marrow cells. To test the effect of abrogating STAT3 activity in HSCs, primitive murine fetal liver cells were transduced at high efficiency with either a bicistronic dominant-negative (dn) or wild-type (wt) STAT3-IRES-GFP retrovirus. Dn STAT3-transduced HSCs showed markedly and permanently reduced in vivo lympho-myeloid reconstituting ability relative to co-transplanted non-transduced HSCs or HSCs transduced with a control (GFP-only) vector. In contrast, the activity of dn STAT3-transduced cells with short term in vivo (CFU-S) or in vitro (CFC) proliferation potential was not affected. Overexpression of wt-STAT3 had very little effect on either HSCs or shorter term progenitors. These findings suggest HSCs express non-limiting levels of STAT3 which, nevertheless, play an important stage-specific and non-redundant role in maintaining the function of HSCs stimulated to divide in adult marrow tissue.

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References

  • Audet J, Miller CL, Rose-John S, Piret J, Eaves CJ . 2001 Proc. Natl. Acad. Sci. USA 98: 1757–1762

  • Bernad A, Kopf M, Kulbacki R, Weich N, Koehler G, Gutierrez-Ramos JC . 1994 Immunity 1: 725–731

  • Biethahn S, Alves F, Wilde S, Hiddeman W, Spiekermann K . 1999 Exp. Hematol. 27: 885–894

  • Caldenhoven E, van Dijk TB, Solari R, Armstrong J, Raaijmakers JAM, Lammers W-WJ, Koenderman L, de Groot RP . 1996 J. Biol. Chem. 22: 13221–13227

  • Chakraborty A, Tweardy DJ . 1998 Leuk. Lymphoma 30: 433–442

  • Chakraborty A, White SM, Schaefer TS, Ball ED, Dyer KF, Tweardy DJ . 1996 Blood 88: 2442–2449

  • Cheng T, Rodrigues N, Shen H, Yang Y-G, Dombkowski D, Sykes M, Scadden DT . 2000 Science 287: 1804–1808

  • Conneally E, Cashman J, Petzer A, Eaves C . 1997 Proc. Natl. Acad. Sci. USA 94: 9836–9841

  • Darnell Jr JE . 1997 Science 277: 1630–1635

  • Fraser CC, Szilvassy SJ, Eaves CJ, Humphries RK . 1992 Proc. Natl. Acad. Sci. USA 89: 1968–1972

  • Hennemann B, Chuo JY, Schley PD, Lambie K, Humphries RK, Eaves CJ . 2000 Hum. Gene Ther. 11: 43–51

  • Holyoake T, Jiang X, Eaves C, Eaves A . 1999 Blood 94: 2056–2064

  • Kishimoto A, Akira S, Narazaki M, Taga T . 1995 Blood 86: 1243–1254

  • Lemieux ME, Rebel VI, Lansdorp PM, Eaves CJ . 1995 Blood 86: 1339–1347

  • Markowitz D, Goff S, Bank A . 1988 J. Virol. 62: 1120–1124

  • Matsuda T, Nakamura T, Nakao K, Arai T, Katsuki M, Heike T, Yokota T . 1999 EMBO J. 18: 4261–4269

  • Minami M, Inoue M, Wei S, Takeda K, Matsumoto M, Kishimoto T, Akira S . 1996 Proc. Natl. Acad. Sci. USA 93: 3963–3966

  • Niwa M, Burdon T, Chambers I, Smith A . 1998 Genes Dev. 12: 2048–2060

  • O'Farrell A-M, Liu Y, Moore KW, Mui AL-F . 1998 EMBO J. 17: 1006–1018

  • Oh I-H, Lau A, Eaves CJ . 2000 Blood 96: 4160–4168

  • Peters M, Schirmacher P, Goldschmitt J, Odenthal M, Peschel C, Dienes H-P, Fattori E, Ciliberto G, Meyer zum Buschenfelde KH, Rose-John S . 1997 J. Exp. Med. 185: 755–766

  • Petzer AL, Hogge DE, Lansdorp PM, Reid DS, Eaves CJ . 1996 Proc. Natl. Acad. Sci. USA 93: 1470–1474

  • Raz R, Lee CK, Cannizzarro LA, d'Eustachio P, Levy DE . 1999 Proc. Natl. Acad. Sci. USA 96: 2846–2851

  • Rebel VI, Lansdorp PM . 1996 J. Hematother. 5: 25–37

  • Rebel VI, Miller CL, Eaves CJ, Lansdorp PM . 1996 Blood 87: 3500–3507

  • Sasse J, Hemmann U, Schwartz C, Schniertshauer U, Heesel B, Landgraf C, Schneider-Mergener J, Heinrich P, Horn F . 1997 Mol. Cell. Biol. 17: 4677–4686

  • Schaefer TS, Sanders LK, Nathans D . 1995 Proc. Natl. Acad. Sci. USA 92: 9097–9101

  • Szilvassy SJ, Humphries RK, Lansdorp PM, Eaves AC, Eaves CJ . 1990 Proc. Natl. Acad. Sci. USA 87: 8736–8740

  • Szilvassy SJ, Nicolini FE, Eaves CJ, Miller CL . 2001 Methods in Molecular Medicine: Hematopoietic Stem Cell Protocols: Quantitation of murine and human hematopoietic stem cells by limiting-dilution analysis in competitively repopulated hosts Jordon CT and Klug CA (eds) Totowa, NJ: Humana Press pp. 167–187

    Google Scholar 

  • Taga T, Kishimoto T . 1997 Annu. Rev. Immunol. 15: 797–819

  • Thomas TE, Miller CL, Eaves CJ . 1999 Methods: A Companion to Methods in Enzymology: Purification of hematopoietic stem cells for further biological study Vol 17: New York: Academic Press pp 202–218

    Google Scholar 

  • Wang JCY, Doedens M, Dick JE . 1997 Blood 89: 3919–3924

  • Yoshida K, Taga T, Saito M, Suematsu S, Kumanogoh A, Tanaka T, Fujiwara H, Hirata M, Yamagami T, Nakahata T, Hirabayashi T, Yoneda Y, Tanaka K, Wang W-Z, Mori C, Shiota K, Yoshida N, Kishimoto T . 1996 Proc. Natl. Acad. Sci. USA 93: 407–411

  • Zandstra PW, Conneally E, Petzer AL, Piret JM, Eaves CJ . 1997 Proc. Natl. Acad. Sci. USA 94: 4698–4703

  • Zhang X, Blenis J, Li HC, Schindler C, Chen-Kiang S . 1995 Science 267: 1990–1994

  • Zong CS, Zeng L, Jiang Y, Sadowski HB, Wang LH . 1998 J. Biol. Chem. 273: 28065–28072

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Acknowledgements

The authors thank Cindy Cao, Geoff Gotto, Maya Sinclair, Margaret Hale, and the staff of the Terry Fox Laboratory FACS service for expert technical assistance, Dr A Mui for the STAT3 plasmids, Dr K Humphries for the MSCV–IRES–GFP plasmid and Amy Ahamed for typing. This study was supported by the National Cancer Institute of Canada (NCIC) with funds from the Canadian Cancer Society and the Terry Fox Run. I-H Oh held a NCIC Postdoctoral Fellowship and C Eaves was a Terry Fox Cancer Research Scientist of the NCIC.

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  1. Il-Hoan Oh

    Present address: Catholic Institute for Medical Science, Catholic University of Korea, Seoul, Korea

Authors and Affiliations

  1. British Columbia Cancer Agency and Department of Medical Genetics, Terry Fox Laboratory, University of British Columbia, Vancouver, V5Z 1L3, British Columbia, Canada

    Il-Hoan Oh & Connie J Eaves

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  2. Connie J Eaves
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Correspondence to Connie J Eaves.

Appendix

Appendix

Inhibition of endogenous STAT3 activation by dn STAT3 in stably transduced M1 cells. Aliquots of cells stably-transduced with each retroviral vector were stimulated with IL-6 (100 ng/ml) for 15 min and then analysed by Western blotting for the presence of phosphorylated STAT3 using an anti-phosphotyrosine (Tyr705)-specific anti-STAT3 antibody (105 cells per lane). The membrane was then stripped and re-probed for total STAT3 protein using a polyclonal antibody against STAT3 (see Figure 10).

figure 10

Figure 10

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Oh, IH., Eaves, C. Overexpression of a dominant negative form of STAT3 selectively impairs hematopoietic stem cell activity. Oncogene 21, 4778–4787 (2002). https://doi.org/10.1038/sj.onc.1205592

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