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Disruption of the PRKCD–FBXO25–HAX-1 axis attenuates the apoptotic response and drives lymphomagenesis

Nature Medicine volume 20, pages 1401–1409 (2014)Cite this article

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Abstract

We searched for genetic alterations in human B cell lymphoma that affect the ubiquitin-proteasome system. This approach identified FBXO25 within a minimal common region of frequent deletion in mantle cell lymphoma (MCL). FBXO25 encodes an orphan F-box protein that determines the substrate specificity of the SCF (SKP1–CUL1–F-box)FBXO25 ubiquitin ligase complex. An unbiased screen uncovered the prosurvival protein HCLS1-associated protein X-1 (HAX-1) as the bona fide substrate of FBXO25 that is targeted after apoptotic stresses. Protein kinase Cδ (PRKCD) initiates this process by phosphorylating FBXO25 and HAX-1, thereby spatially directing nuclear FBXO25 to mitochondrial HAX-1. Our analyses in primary human MCL identify monoallelic loss of FBXO25 and stabilizing HAX1 phosphodegron mutations. Accordingly, FBXO25 re-expression in FBXO25-deleted MCL cells promotes cell death, whereas expression of the HAX-1 phosphodegron mutant inhibits apoptosis. In addition, knockdown of FBXO25 significantly accelerated lymphoma development in Eμ-Myc mice and in a human MCL xenotransplant model. Together we identify a PRKCD-dependent proapoptotic mechanism controlling HAX-1 stability, and we propose that FBXO25 functions as a haploinsufficient tumor suppressor and that HAX1 is a proto-oncogene in MCL.

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Figure 1: FBXO25 regulates the apoptotic response by targeting HAX-1 for degradation.
Figure 2: Phosphorylation of both FBXO25and HAX-1 determines spatial and temporal control of HAX-1 degradation.
Figure 3: PRKCD phosphorylates FBXO25 and HAX-1 to promote apoptosis through HAX-1 degradation.
Figure 4: FBXO25 suppresses lymphoma growth and development through HAX-1 stabilization in vivo.
Figure 5: Disruption of the PRKCD–FBXO25–HAX-1 axis promotes survival in MCL.
Figure 6: Identification of monoallelic FBXO25 deletions and stabilizing HAX1 degron mutations in primary human MCL samples.

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Acknowledgements

We thank J. Ihle (St. Jude Children's Research Hospital), V. Kalscheuer (Max Planck Institute for Molecular Genetics), M. Pagano (New York University School of Medicine) and A. Villunger (Medical University of Innsbruck) for reagents and cell lines; J. Martinez-Climent, M. Schraders, H. Tagawa, H. Kohlhammer and E. Flordal Thelander for sending the raw data of aCGH studies; G. Keller and K. Malinowski for suggestions; K.-F. Becker and C. Schott for help with extract preparation from MCL tissue samples; and R. Oellinger for help with the MCL tissue culture models. This work was supported by grants from the German Research Foundation (KE 222/-1 and SFB824 to U.K.; and BA 2851/4-1 and Emmy Noether Program grant BA 2851/3-1 to F.B.) and the German Cancer Aid (#109543 to F.B.).

Author information

Author notes

  1. Ursula Baumann and Vanesa Fernández-Sáiz: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine III, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany

    Ursula Baumann, Vanesa Fernández-Sáiz, Anna-Maria Knorn, Katharina Engel, Viktoriya Tomiatti, Bianca-Sabrina Targosz, Christian Peschel, Ulrich Keller & Florian Bassermann

  2. Institute of Pathology, Julius-Maximilians-University and Comprehensive Cancer Center Mainfranken, Wuerzburg, Germany

    Martina Rudelius & Andreas Rosenwald

  3. Department of Proteomics and Bioanalytics, Technische Universität München, Freising, Germany

    Simone Lemeer & Bernhard Kuster

  4. Department of Medicine II, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany

    Roland Rad

  5. German Cancer Consortium (DKTK) and German Cancer Research Center (DKFZ), Heidelberg, Germany

    Roland Rad, Christian Peschel, Ulrich Keller, Bernhard Kuster & Florian Bassermann

  6. Department of Nuclear Medicine, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany

    Jolanta Slawska & Zhoulei Li

  7. Department of Gene Vectors, Helmholtz Center Munich–German Research Center for Environmental Health, Munich, Germany

    Irmela Jeremias

  8. Department of Pediatrics, Dr. von Hauner Children's Hospital, Ludwig Maximilians University, Munich, Germany

    Irmela Jeremias

  9. Department of Hematology/Oncology, University Medical Center Freiburg, Freiburg, Germany

    Anna-Lena Illert & Cornelius Miething

  10. Department of Prostate Cancer Research, Institute of Pathology, University Hospital of Bonn, Bonn, Germany

    Martin Braun & Sven Perner

  11. Biotechnology Centre of Oslo, Oslo, Norway

    Michael Leitges

  12. Institute of Pathology, Hematopathology Section and Lymph Node Registry, Universitätsklinikum Schleswig-Holstein, Kiel, Germany

    Wolfram Klapper

  13. Department of Medicine III, University Hospital Munchen, Munich, Germany

    Martin Dreyling

  14. Department of Medicine A, Translational Oncology, Universitätsklinikum Münster, Münster, Germany

    Georg Lenz

  15. Center for Integrated Protein Science Munich (CIPSM), Freising, Germany

    Bernhard Kuster

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  1. Ursula Baumann
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Contributions

U.B., V.F.-S. and F.B. conceived and designed the research. U.B. and V.F.-S. performed most of the experiments with crucial help from A.-M.K., B.-S.T., V.T. and K.E. I.J., J.S. and Z.L. helped with the MCL xenotransplant model. M.R., A.R., W.K. and M.D. provided lymphoma samples. M.R. and A.R. performed IHC analyses. R.R. performed analyses of aCGH data. S.L. and B.K. performed mass spectrometry. C.M., A.-L.I., G.L., M.B., S.P. and M.L. provided critical reagents. U.B., V.F.-S., B.-S.T., K.E., S.L., I.J., C.P., U.K., B.K. and F.B. analyzed results. F.B. coordinated this work and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Florian Bassermann.

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Baumann, U., Fernández-Sáiz, V., Rudelius, M. et al. Disruption of the PRKCD–FBXO25–HAX-1 axis attenuates the apoptotic response and drives lymphomagenesis. Nat Med 20, 1401–1409 (2014). https://doi.org/10.1038/nm.3740

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