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γ-secretase inhibitors reverse glucocorticoid resistance in T cell acute lymphoblastic leukemia

Nature Medicine volume 15, pages 5058 (2009) | Download Citation

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Abstract

Gamma-secretase inhibitors (GSIs) block the activation of the oncogenic protein Notch homolog-1 (NOTCH1) in T cell acute lymphoblastic leukemia (T-ALL). However, limited antileukemic cytotoxicity and severe gastrointestinal toxicity have restricted the clinical application of these targeted drugs. Here we show that combination therapy with GSIs plus glucocorticoids can improve the antileukemic effects of GSIs and reduce their gut toxicity in vivo. Inhibition of NOTCH1 signaling in glucocorticoid-resistant T-ALL restored glucocorticoid receptor autoupregulation and induced apoptotic cell death through induction of the gene encoding BCL-2–like apoptosis initiator-11 (BCL2L11). GSI treatment resulted in cell cycle arrest and accumulation of goblet cells in the gut mediated by upregulation of the gene encoding the transcription factor Krüppel-like factor-4 (Klf4), a negative regulator of the cell cycle required for goblet cell differentiation. In contrast, glucocorticoid treatment induced transcriptional upregulation of cyclin D2 (Ccnd2) and protected mice from developing the intestinal goblet cell metaplasia typically induced by inhibition of NOTCH signaling with GSIs. These results support a role for glucocorticoids plus GSIs in the treatment of glucocorticoid-resistant T-ALL.

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Acknowledgements

We thank U. Klein for help with flow cytometry analysis; T. Matos for the processing of histological, histochemical and imunohistochemical assays; P. Sicinski (Dana-Farber Cancer Institute) for the Ccnd2-knockout mouse; T. Honjo (Kyoto University) and B. Reitzis (Columbia University) for the Rbpjflox/flox conditional knockout mouse; T. Ludwig (Columbia University) for the Cre-Tam mouse line; A. Kung (Dana-Farber Cancer Institute) for the FUW-Luc neo vector; J. Aster (Brigham and Women's Hospital) for the MigR1 and MigR1-ICN1 retroviral constructs and for the NOTCH1-TAD antibody; W. Hahn (Dana-Farber Cancer Institute) for the pLKO shRNA vector; R. Dalla Favera (Columbia University) for the pCM8-cMYB vector; M.J. Garabedian (New York University) for the pCMV-HA-hGR vector; and N. Wu and C. Wa for pharmacokinetic and drug metabolism analysis. This work was supported by the US National Institutes of Health (R01CA120196 and R01CA129382 to A.F. and R56AI070310, 1R01CA105129 and 1R01CA133379 to I.A.); the Wipe Out Leukemia Forever Foundation (A.F.), the Leukemia and Lymphoma Society (grants 1287-08 and 6237-08 to A.F.), the Charlotte Geyer Foundation (A.F.), the Cancer Research Institute (A.F.), the Swim Across America Foundation (A.F.), the Golfers Against Cancer Foundation (A.F.), the American Cancer Society (RSG0806801 to I.A.), the Città della Speranza Foundation, a Cassa di Risparmio di Padova e Rovigo research grant, a Programmi di Ricerca Scientifica di Rilevante Interesse Nazionale, and the Fondo de Investigacion Sanitaria (grant CD07/00033 to P.J.R.). A.F. and I.A. are Leukemia & Lymphoma Society Scholars. T.P. is a recipient of a Young Investigator Award from the Alex's Lemonade Stand Foundation. I.H. is supported by the Dutch Cancer Society.

Author information

Author notes

    • Valeria Tosello
    •  & Teresa Palomero

    These authors contributed equally to this work.

Affiliations

  1. Institute for Cancer Genetics, Columbia University, 1130 St. Nicholas Avenue, New York, New York 10032, USA.

    • Pedro J Real
    • , Valeria Tosello
    • , Teresa Palomero
    • , Maria Luisa Sulis
    • , Kelly Barnes
    •  & Adolfo Ferrando
  2. Andalusian Stem Cell Bank, Centro de Investigación Biomédica, Avenida del Conocimiento, Granada 18100, Spain.

    • Pedro J Real
  3. Department of Pathology, Columbia University Medical Center, 630 West 168th Street, New York, New York 10032, USA.

    • Teresa Palomero
    • , Mireia Castillo
    • , Carlos Cordon-Cardo
    •  & Adolfo Ferrando
  4. Department of Pathology, New York University Medical Center, 550 First Avenue, New York, New York 10016, USA.

    • Eva Hernando
  5. Antitumor Assessment Core Facility, Memorial Sloan-Kettering Cancer Center, Zuckerman Research Center, 417 East 68th Street, New York, New York 10021, USA.

    • Elisa de Stanchina
  6. Department of Pediatrics, Columbia University Medical Center, 161 Fort Washington Avenue, New York, New York 10032, USA.

    • Maria Luisa Sulis
    •  & Adolfo Ferrando
  7. Department of Pathology, New York University Cancer Institute, 530 First Avenue, New York, New York 10016, USA.

    • Catherine Sawai
    •  & Iannis Aifantis
  8. Department of Pediatric Oncology/Hematology, Erasmus MC-Sophia Children's Hospital, Dr. Molewaterplein 60, Rotterdam 3015GJ, The Netherlands.

    • Irene Homminga
    •  & Jules Meijerink
  9. Laboratory of Oncohematology, Department of Pediatrics `Salus Pueri', University of Padua, Via Giustiniani 3, Padua 35128, Italy.

    • Giuseppe Basso
  10. Department of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine, 6311 Garners Ferry Road, Columbia, South Carolina 29208, USA.

    • Walden Ai

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Contributions

A.F. supervised the project. P.J.R., V.T., T.P., M.L.S, K.B., C.S. and I.H. designed and performed experiments. M.C. performed histological and immunohistochemical studies. E.H. and E.d.S. assisted in bioimaging analysis of tumor xenografts. G.B. provided clinical samples. J.M. provided clinical samples and supervised drug sensitivity assays in these cells. A.F. and I.A. supervised the analysis of Ccnd2−/− mice. C.C.-C. supervised histological and immunohistochemical studies. W.A. designed and performed experiments on the regulation of Klf4 by NOTCH1 and HES1. A.F. wrote the manuscript.

Corresponding author

Correspondence to Adolfo Ferrando.

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    Supplementary Text and Figures

    Supplementary Results and Discussion, Supplementary Methods and Supplementary Figs. 1–17

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DOI

https://doi.org/10.1038/nm.1900

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