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NOTCH1 promotes T cell leukemia-initiating activity by RUNX-mediated regulation of PKC-θ and reactive oxygen species

Nature Medicine volume 18, pages 1693–1698 (2012)Cite this article

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Abstract

Reactive oxygen species (ROS), a byproduct of cellular metabolism, damage intracellular macromolecules and, when present in excess, can promote normal hematopoietic stem cell differentiation and exhaustion1,2,3. However, mechanisms that regulate the amount of ROS in leukemia-initiating cells (LICs) and the biological role of ROS in these cells are largely unknown. We show here that the ROSlow subset of CD44+ cells in T cell acute lymphoblastic leukemia (T-ALL), a malignancy of immature T cell progenitors, is highly enriched in the most aggressive LICs and that ROS accumulation is restrained by downregulation of protein kinase C θ (PKC-θ). Notably, primary mouse T-ALLs lacking PKC-θ show improved LIC activity, whereas enforced PKC-θ expression in both mouse and human primary T-ALLs compromised LIC activity. We also show that PKC-θ is regulated by a new pathway in which NOTCH1 induces runt-related transcription factor 3 (RUNX3), RUNX3 represses RUNX1 and RUNX1 induces PKC-θ. NOTCH1, which is frequently activated by mutation in T-ALL4,5,6 and required for LIC activity in both mouse and human models7,8, thus acts to repress PKC-θ. These results reveal key functional roles for PKC-θ and ROS in T-ALL and suggest that aggressive biological behavior in vivo could be limited by therapeutic strategies that promote PKC-θ expression or activity, or the accumulation of ROS.

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Figure 1: LICs in T-ALL are characterized by low amounts of ROS.
Figure 2: PKC-θ dictates the amount of ROS and is asymmetrically distributed to LIC-depleted cell fractions.
Figure 3: Increased PKC-θ expression compromises disease reconstitution in vivo.
Figure 4: NOTCH1 represses PKC-θ and ROS through RUNX3 and RUNX1.

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Acknowledgements

We would like to thank Z. Sun (City of Hope, Duarte, California) for providing PKC-θ knockout mice, J. Fletcher and W. Ou (Brigham and Women's Hospital, Boston, Massachusetts) for PKC-θ shRNA lentivectors, E. Kieff (Brigham and Women's Hospital, Boston, Massachusetts) for CSL antibody, I. Bernstein (Fred Hutchinson Cancer Research Center, Seattle, Washington) for Ig-DL1 ligand, F. Pflumio (Commissariat à l'Énergie Atomique Fontenay-aux-Roses, France) for the MS5-DL1 cell line and E. Martignani for help with immunofluorescence. We would also like to thank P. Ballerini (Hôpital Armand Trousseau, Paris, France), T. Leblanc (Hôpital Saint-Louis, Paris, France), L. Matherly (Karmanos Cancer Institute, Detroit, Michigan) and M.J. You (MD Anderson Cancer Center, Houston, Texas) for providing patient T-ALL samples. This work was supported by grants from the Canadian Institutes of Health Research/Terry Fox Foundation, the Leukemia and Lymphoma Society of Canada, the Cancer Research Society, the Lymphoma Foundation Canada (to A.P.W.) and the US National Cancer Institute (P01CA119070 to J.C.A.). M.Y.C. was supported by a career development award from the US National Cancer Institute (K08CA120544). A.P.W. was a Michael Smith Foundation for Health Research Scholar.

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Authors and Affiliations

  1. Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, British Columbia, Canada

    Vincenzo Giambra, Christopher R Jenkins, Sonya H Lam, Olena O Shevchuk, Oksana Nemirovsky, Carol Wai, Sam Gusscott, R Keith Humphries, Connie Eaves & Andrew P Weng

  2. Department of Pathology, Brigham and Women's Hospital/Harvard Medical School, Boston, Massachusetts, USA.,

    Hongfang Wang & Jon C Aster

  3. Division of Hematology/Oncology, University of Michigan Comprehensive Cancer Center, Ann Arbor, Michigan, USA

    Mark Y Chiang

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Contributions

V.G. and A.P.W. designed the experiments. V.G., C.R.J., S.H.L., O.O.S., O.N., C.W. and S.G. generated the data. V.G. and A.P.W. interpreted the results. M.Y.C. provided reagents and advice. H.W. and J.C.A. generated and analyzed ChIP-Seq data. R.K.H. and C.E. provided advice and discussion. V.G. and A.P.W. wrote the paper.

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Correspondence to Andrew P Weng.

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Giambra, V., Jenkins, C., Wang, H. et al. NOTCH1 promotes T cell leukemia-initiating activity by RUNX-mediated regulation of PKC-θ and reactive oxygen species. Nat Med 18, 1693–1698 (2012). https://doi.org/10.1038/nm.2960

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