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Expression of constitutively active Notch4 (Int-3) modulates myeloid proliferation and differentiation and promotes expansion of hematopoietic progenitors

Leukemia volume 18, pages 777–787 (2004)Cite this article

Abstract

The Notch family of transmembrane receptors has been implicated in the regulation of many developmental processes. In this study, we evaluated the role of Notch4 in immature hematopoietic progenitors by inducing, with retroviral transduction, enforced expression of Int-3, the oncogenic and constitutively active form of mouse Notch4. Int-3-transduced human myeloid leukemia (HL-60) cells demonstrated significantly delayed expression of differentiation markers following retinoic acid and 12-0-tetradecanoylphorbol 13-acetate treatment. Furthermore, HL-60 cells expressing Int-3 displayed a slower growth rate than cells infected with void virus, and accumulation in the G0/G1 phases of cell cycle. Transduction with deletion mutants of Int-3 defined the importance of individual domains of the protein (in particular, the ANK domain and the C-terminal domain) in the inhibition of differentiation and growth arrest of HL-60 cells. When mouse bone marrow enriched for stem cells (5-fluorouracil-resistant, lineage negative) was transduced and cultured for two weeks, the Int-3-transduced population displayed a lower expression of differentiation markers and a three- to five-fold higher frequency of colony-forming cells (CFU-GM/BFU-E) than control cultures. These results strongly support the notion that Notch signaling inhibits differentiation and promotes expansion of hematopoietic stem/progenitor cells.

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Acknowledgements

We wish to thank Kate deBeer for her secretarial assistance in preparation of this manuscript. This work was supported by NIH Grant HL61401 and NCI Grant PO1CA59350 (to MASM) and CNR Biotechnology Programme (to GM).

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Authors and Affiliations

  1. James Ewing Laboratory of Developmental Hematopoiesis, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

    Q Ye, J-H Shieh, G Morrone & M A S Moore

  2. Department of Experimental and Clinical Medicine, University of Catanzaro ‘Magna Graecia’, Catanzaro, Italy

    G Morrone

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Correspondence to M A S Moore.

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Ye, Q., Shieh, JH., Morrone, G. et al. Expression of constitutively active Notch4 (Int-3) modulates myeloid proliferation and differentiation and promotes expansion of hematopoietic progenitors. Leukemia 18, 777–787 (2004). https://doi.org/10.1038/sj.leu.2403291

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