Abstract

Similar to resting mature B cells, where the B-cell antigen receptor (BCR) controls cellular survival1,2,3, surface BCR expression is conserved in most mature B-cell lymphomas. The identification of activating BCR mutations and the growth disadvantage upon BCR knockdown of cells of certain lymphoma entities has led to the view that BCR signalling is required for tumour cell survival4,5,6,7. Consequently, the BCR signalling machinery has become an established target in the therapy of B-cell malignancies8,9. Here we study the effects of BCR ablation on MYC-driven mouse B-cell lymphomas and compare them with observations in human Burkitt lymphoma. Whereas BCR ablation does not, per se, significantly affect lymphoma growth, BCR-negative (BCR) tumour cells rapidly disappear in the presence of their BCR-expressing (BCR+) counterparts in vitro and in vivo. This requires neither cellular contact nor factors released by BCR+ tumour cells. Instead, BCR loss induces the rewiring of central carbon metabolism, increasing the sensitivity of receptor-less lymphoma cells to nutrient restriction. The BCR attenuates glycogen synthase kinase 3 beta (GSK3β) activity to support MYC-controlled gene expression. BCR tumour cells exhibit increased GSK3β activity and are rescued from their competitive growth disadvantage by GSK3β inhibition. BCR lymphoma variants that restore competitive fitness normalize GSK3β activity after constitutive activation of the MAPK pathway, commonly through Ras mutations. Similarly, in Burkitt lymphoma, activating RAS mutations may propagate immunoglobulin-crippled tumour cells, which usually represent a minority of the tumour bulk. Thus, while BCR expression enhances lymphoma cell fitness, BCR-targeted therapies may profit from combinations with drugs targeting BCR tumour cells.

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References

  1. 1.

    , & In vivo ablation of surface immunoglobulin on mature B cells by inducible gene targeting results in rapid cell death. Cell 90, 1073–1083 (1997)

  2. 2.

    , , & Survival of resting mature B lymphocytes depends on BCR signaling via the Igα/β heterodimer. Cell 117, 787–800 (2004)

  3. 3.

    et al. PI3 kinase signals BCR-dependent mature B cell survival. Cell 139, 573–586 (2009)

  4. 4.

    et al. Chronic active B-cell-receptor signalling in diffuse large B-cell lymphoma. Nature 463, 88–92 (2010)

  5. 5.

    et al. Burkitt lymphoma pathogenesis and therapeutic targets from structural and functional genomics. Nature 490, 116–120 (2012)

  6. 6.

    , , & B-cell receptor signaling in diffuse large B-cell lymphoma. Semin. Hematol. 52, 77–85 (2015)

  7. 7.

    et al. The B cell antigen receptor and overexpression of MYC can cooperate in the genesis of B cell lymphomas. PLoS Biol. 6, e152 (2008)

  8. 8.

    & Targeting pathological B cell receptor signalling in lymphoid malignancies. Nat. Rev. Drug Discov. 12, 229–243 (2013)

  9. 9.

    et al. Targeting B cell receptor signaling with ibrutinib in diffuse large B cell lymphoma. Nat. Med. 21, 922–926 (2015)

  10. 10.

    et al. Burkitt lymphoma in the mouse. J. Exp. Med. 192, 1183–1190 (2000)

  11. 11.

    et al. Synergy between PI3K signaling and MYC in Burkitt lymphomagenesis. Cancer Cell 22, 167–179 (2012)

  12. 12.

    & Vagaries of conditional gene targeting. Nat. Immunol. 8, 665–668 (2007)

  13. 13.

    et al. Survival of Igα-deficient mature B cells requires BAFF-R function. J. Immunol. 196, 2348–2360 (2016)

  14. 14.

    & Pulsed stable isotope-resolved metabolomic studies of cancer cells. Methods Enzymol. 543, 179–198 (2014)

  15. 15.

    et al. The B cell antigen receptor activates the Akt (protein kinase B)/glycogen synthase kinase-3 signaling pathway via phosphatidylinositol 3-kinase. J. Immunol. 163, 1894–1905 (1999)

  16. 16.

    & The renaissance of GSK3. Nat. Rev. Mol. Cell Biol. 2, 769–776 (2001)

  17. 17.

    et al. Selective glycogen synthase kinase 3 inhibitors potentiate insulin activation of glucose transport and utilization in vitro and in vivo. Diabetes 52, 588–595 (2003)

  18. 18.

    et al. The B-cell antigen receptor integrates adaptive and innate immune signals. Proc. Natl Acad. Sci. USA 112, 12145–12150 (2015)

  19. 19.

    et al. Elucidation of tonic and activated B-cell receptor signaling in Burkitt’s lymphoma provides insights into regulation of cell survival. Proc. Natl Acad. Sci. USA 113, 5688–5693 (2016)

  20. 20.

    et al. Selective transcriptional regulation by Myc in cellular growth control and lymphomagenesis. Nature 511, 488–492 (2014)

  21. 21.

    et al. Activating K-Ras mutations outwith ‘hotspot’ codons in sporadic colorectal tumours – implications for personalised cancer medicine. Br. J. Cancer 102, 693–703 (2010)

  22. 22.

    et al. Emergence of KRAS mutations and acquired resistance to anti-EGFR therapy in colorectal cancer. Nature 486, 532–536 (2012)

  23. 23.

    , , & Ras activation of Erk restores impaired tonic BCR signaling and rescues immature B cell differentiation. J. Exp. Med. 207, 607–621 (2010)

  24. 24.

    et al. Activation of Ras overcomes B-cell tolerance to promote differentiation of autoreactive B cells and production of autoantibodies. Proc. Natl Acad. Sci. USA 111, E2797–E2806 (2014)

  25. 25.

    et al. Gene essentiality profiling reveals gene networks and synthetic lethal interactions with oncogenic Ras. Cell 168, 890–903 (2017)

  26. 26.

    , , , & Oncogenic mechanisms in Burkitt lymphoma. Cold Spring Harb. Perspect. Med. 4, a014282 (2014)

  27. 27.

    et al. B cell receptor signal strength determines B cell fate. Nat. Immunol. 5, 317–327 (2004)

  28. 28.

    , , & Ability of the hydrophobic FGF and basic TAT peptides to promote cellular uptake of recombinant Cre recombinase: a tool for efficient genetic engineering of mammalian genomes. Proc. Natl Acad. Sci. USA 99, 4489–4494 (2002)

  29. 29.

    et al. DNA damage response activation in mouse embryonic fibroblasts undergoing replicative senescence and following spontaneous immortalization. Cell Cycle 7, 3601–3606 (2008)

  30. 30.

    et al. Interplay of p53 and DNA-repair protein XRCC4 in tumorigenesis, genomic stability and development. Nature 404, 897–900 (2000)

  31. 31.

    et al. Novel targeted deregulation of c-Myc cooperates with Bcl-X(L) to cause plasma cell neoplasms in mice. J. Clin. Invest. 113, 1763–1773 (2004)

  32. 32.

    et al. Germinal center dysregulation by histone methyltransferase EZH2 promotes lymphomagenesis. J. Clin. Invest. 123, 5009–5022 (2013)

  33. 33.

    et al. An automated GCxGC-TOF-MS protocol for batch-wise extraction and alignment of mass isotopomer matrixes from differential 13C-labelling experiments: a case study for photoautotrophic-mixotrophic grown Chlamydomonas reinhardtii cells. J. Basic Microbiol. 49, 82–91 (2009)

  34. 34.

    , , & Decoding the dynamics of cellular metabolism and the action of 3-bromopyruvate and 2-deoxyglucose using pulsed stable isotope-resolved metabolomics. Cancer Metab. 2, 9 (2014)

  35. 35.

    et al. GMD@CSB.DB: the Golm Metabolome Database. Bioinformatics 21, 1635–1638 (2005)

  36. 36.

    , & Maui-VIA: a user-friendly software for visual identification, alignment, correction, and quantification of gas chromatography-mass spectrometry data. Front. Bioeng. Biotechnol. 2, 84 (2015)

  37. 37.

    et al. Enrichr: interactive and collaborative HTML5 gene list enrichment analysis tool. BMC Bioinformatics 14, 128 (2013)

  38. 38.

    et al. Enrichr: a comprehensive gene set enrichment analysis web server 2016 update. Nucleic Acids Res. 44 (W1), W90–W97 (2016)

  39. 39.

    , , , & SAVI: a statistical algorithm for variant frequency identification. BMC Syst. Biol. 7 (Suppl. 2), S2 (2013)

  40. 40.

    et al. Cutaneous distribution of plasmacytoid dendritic cells in lupus erythematosus. Selective tropism at the site of epithelial apoptotic damage. Immunobiology 214, 877–886 (2009)

  41. 41.

    et al. Recurrent mutation of the ID3 gene in Burkitt lymphoma identified by integrated genome, exome and transcriptome sequencing. Nat. Genet. 44, 1316–1320 (2012)

  42. 42.

    et al. Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia 17, 2257–2317 (2003)

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Acknowledgements

We thank members of the Casola laboratory, A. Ciliberto, K. L. Otipoby and R. Küppers for discussions; A. Ciliberto and N. Bolli for reading the manuscript; D. Wang, M. Caganova, L. Duarte and S. Ronzoni for preliminary work and technical assistance; F. D’Adda di Fagagna for reagents. We acknowledge the German International Cancer Genome Consortium (ICGC) Molecular Mechanisms in Malignant Lymphoma by Sequencing (MMML-Seq) and Molecular Mechanisms in Malignant Lymphoma (MMML) networks. This work was supported by the Italian Association for Cancer Research, the Giovanni Armenise/Harvard Foundation (to S.C.), the National Institutes of Health (to K.R.) and the European Research Council (to S.C. through FIRB IDEAS, and to K.R., Advanced Grant 268921). R.S. was supported by ICGC MMML-Seq grant 01KU1002, ICGC-DE-Mining grant 01KU1505G and e:BIO MMML-MYC-SYS grant 036166, as well as by the KinderKrebsInitiative Buchholz/Holm-Seppensen. G.V. and V.P were supported by the U. Veronesi Foundation. S.L. and M.B. were supported by Fondazione Beretta.

Author information

Author notes

    • Gabriele Varano
    •  & Simon Raffel

    These authors contributed equally to this work.

Affiliations

  1. IFOM, the FIRC Institute of Molecular Oncology, 20139 Milan, Italy

    • Gabriele Varano
    • , Martina Sormani
    • , Federica Zanardi
    • , Laura Perucho
    • , Valentina Petrocelli
    •  & Stefano Casola
  2. Heidelberg Institute for Stem Cell Technology and Experimental Medicine, 69120 Heidelberg, Germany

    • Simon Raffel
  3. Department of Molecular and Translational Medicine, Section of Pathology, University of Brescia, Spedali Civili, 25123 Brescia, Italy

    • Silvia Lonardi
    • , Mattia Bugatti
    •  & Fabio Facchetti
  4. Max-Delbrück-Center of Molecular Medicine in the Helmholtz Association, 13125 Berlin, Germany

    • Christin Zasada
    • , Stefan Kempa
    •  & Klaus Rajewsky
  5. Institute of Human Genetics, Christian-Albrechts-University Kiel, 24105 Kiel, Germany

    • Andrea Haake
    • , Ulrike Paul
    •  & Reiner Siebert
  6. Department of Systems Biology and Department of Biomedical Informatics, Columbia University, New York, New York 10027, USA

    • Albert K. Lee
    •  & Raul Rabadan
  7. Division of Genetics and Cell Biology, San Raffaele Scientific Institute, 20132 Milan, Italy

    • Eelco Van Anken
  8. Institute for Cancer Genetics and the Department of Pathology and Cell Biology, Herbert Irving Comprehensive Cancer Center, Columbia University, New York, New York 10032, USA

    • Laura Pasqualucci
  9. Institute of Human Genetics, University Hospital of Ulm, 89081 Ulm, Germany

    • Reiner Siebert
  10. Ateneo Vita Salute and San Raffaele Scientific Institute, Pathology and Lymphoid Malignancies Units, 20132 Milan, Italy

    • Maurilio Ponzoni

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Contributions

M.S. and F.Z. contributed equally to this work. G.V. performed most experiments. S.R. made initial observations. M.S. performed CAL-101 lymphoma sensitivity assays, in vivo transplantations and biochemical analyses. M.S. analysed Xbp1 expression with E.V.A. F.Z. performed BCR complementation studies and bioinformatics analyses on transcriptome and exome sequencing data. C.Z. and M.S. performed metabolic labelling of lymphomas. C.Z. and S.K. analysed and interpreted metabolomic data. L.Pe. investigated the role of cell contact and soluble factors in the counter-selection of BCR tumour cells. V.P. performed Burkitt lymphoma immunoglobulin gene rearrangement analyses and Ras complementation assays. S.L., M.B., M.P. and F.F. performed immunohistochemical analyses and interpreted the results. A.H., U.P. and R.S. analysed RAS genes in Burkitt lymphoma. A.K.L., L.Pa. and R.R. assisted with exome sequencing analyses. S.C. and K.R. conceived the project. S.C. designed the experiments and interpreted the results with G.V. S.C. wrote and edited the manuscript with help from K.R. and G.V.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Stefano Casola.

Reviewer Information Nature thanks J. Burger, A. Schaffer and A. Thomas-Tikhonenko for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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