Germinal centre B-cell-like diffuse large B-cell lymphoma (GCB-DLBCL) is a common malignancy, yet the signalling pathways that are deregulated and the factors leading to its systemic dissemination are poorly defined1,2. Work in mice showed that sphingosine-1-phosphate receptor-2 (S1PR2), a Gα12 and Gα13 coupled receptor, promotes growth regulation and local confinement of germinal centre B cells3,4. Recent deep sequencing studies of GCB-DLBCL have revealed mutations in many genes in this cancer, including in GNA13 (encoding Gα13) and S1PR2 (refs 5,6, 7). Here we show, using in vitro and in vivo assays, that GCB-DLBCL-associated mutations occurring in S1PR2 frequently disrupt the receptor’s Akt and migration inhibitory functions. Gα13-deficient mouse germinal centre B cells and human GCB-DLBCL cells were unable to suppress pAkt and migration in response to S1P, and Gα13-deficient mice developed germinal centre B-cell-derived lymphoma. Germinal centre B cells, unlike most lymphocytes, are tightly confined in lymphoid organs and do not recirculate. Remarkably, deficiency in Gα13, but not S1PR2, led to germinal centre B-cell dissemination into lymph and blood. GCB-DLBCL cell lines frequently carried mutations in the Gα13 effector ARHGEF1, and Arhgef1 deficiency also led to germinal centre B-cell dissemination. The incomplete phenocopy of Gα13- and S1PR2 deficiency led us to discover that P2RY8, an orphan receptor that is mutated in GCB-DLBCL and another germinal centre B-cell-derived malignancy, Burkitt’s lymphoma, also represses germinal centre B-cell growth and promotes confinement via Gα13. These findings identify a Gα13-dependent pathway that exerts dual actions in suppressing growth and blocking dissemination of germinal centre B cells that is frequently disrupted in germinal centre B-cell-derived lymphoma.

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  1. 1.

    , & Pathogenesis of human B cell lymphomas. Annu. Rev. Immunol. 30, 565–610 (2012)

  2. 2.

    & Germinal centres: role in B-cell physiology and malignancy. Nature Rev. Immunol. 8, 22–33 (2008)

  3. 3.

    et al. The sphingosine 1-phosphate receptor S1P(2) maintains the homeostasis of germinal center B cells and promotes niche confinement. Nature Immunol. 12, 672–680 (2011)

  4. 4.

    et al. Targeted disruption of the S1P2 sphingosine 1-phosphate receptor gene leads to diffuse large B-cell lymphoma formation. Cancer Res. 69, 8686–8692 (2009)

  5. 5.

    et al. Discovery and prioritization of somatic mutations in diffuse large B-cell lymphoma (DLBCL) by whole-exome sequencing. Proc. Natl Acad. Sci. USA 109, 3879–3884 (2012)

  6. 6.

    et al. Frequent mutation of histone-modifying genes in non-Hodgkin lymphoma. Nature 476, 298–303 (2011)

  7. 7.

    et al. Mutational and structural analysis of diffuse large B-cell lymphoma using whole-genome sequencing. Blood 122, 1256–1265 (2013)

  8. 8.

    , & G12/G13-mediated signalling in mammalian physiology and disease. Trends Pharmacol. Sci. 29, 582–589 (2008)

  9. 9.

    et al. The genetic landscape of mutations in Burkitt lymphoma. Nature Genet. 44, 1321–1325 (2012)

  10. 10.

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

  11. 11.

    et al. Genetic heterogeneity of diffuse large B-cell lymphoma. Proc. Natl Acad. Sci. USA 110, 1398–1403 (2013)

  12. 12.

    et al. BCL2 mutations in diffuse large B-cell lymphoma. Leukemia 26, 1383–1390 (2012)

  13. 13.

    et al. Impact of concordant and discordant bone marrow involvement on outcome in diffuse large B-cell lymphoma treated with R-CHOP. J. Clin. Oncol. 29, 1452–1457 (2011)

  14. 14.

    et al. COSMIC: mining complete cancer genomes in the Catalogue of Somatic Mutations in Cancer. Nucleic Acids Res. 39, D945–D950 (2011)

  15. 15.

    , & Pathogenesis of non-Hodgkin’s lymphoma. J. Clin. Oncol. 29, 1803–1811 (2011)

  16. 16.

    et al. Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin. Nature Genet. 42, 181–185 (2010)

  17. 17.

    et al. PTEN loss defines a PI3K/AKT pathway-dependent germinal center subtype of diffuse large B-cell lymphoma. Proc. Natl Acad. Sci. USA 110, 12420–12425 (2013)

  18. 18.

    et al. Lymphoproliferative disease and autoimmunity in mice with increased miR-17–92 expression in lymphocytes. Nature Immunol. 9, 405–414 (2008)

  19. 19.

    et al. EZH2 is required for germinal center formation and somatic EZH2 mutations promote lymphoid transformation. Cancer Cell 23, 677–692 (2013)

  20. 20.

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

  21. 21.

    et al. Analysis of the coding genome of diffuse large B-cell lymphoma. Nature Genet. 43, 830–837 (2011)

  22. 22.

    et al. BCL2 translocation defines a unique tumor subset within the germinal center B-cell-like diffuse large B-cell lymphoma. Am. J. Pathol. 165, 159–166 (2004)

  23. 23.

    , , & Signalling mechanisms of RhoGTPase regulation by the heterotrimeric G proteins G12 and G13. J. Biochem. 150, 357–369 (2011)

  24. 24.

    et al. Recurrent RHOA mutations in pediatric Burkitt lymphoma treated according to the NHL-BFM protocols. Genes Chromosomes Cancer (2014)

  25. 25.

    et al. Regulation of PTEN by Rho small GTPases. Nature Cell Biol. 7, 399–404 (2005)

  26. 26.

    , & Inhibitory role of sphingosine 1-phosphate receptor 2 in macrophage recruitment during inflammation. J. Immunol. 184, 1475–1483 (2010)

  27. 27.

    , & Rho protein crosstalk: another social network? Trends Cell Biol. 21, 718–726 (2011)

  28. 28.

    et al. The sphingosine-1-phosphate receptors S1P1, S1P2, and S1P3 function coordinately during embryonic angiogenesis. J. Biol. Chem. 279, 29367–29373 (2004)

  29. 29.

    , , , & Rho GEF Lsc is required for normal polarization, migration, and adhesion of formyl-peptide-stimulated neutrophils. Blood 107, 1627–1635 (2006)

  30. 30.

    et al. Essential role for Gα13 in endothelial cells during embryonic development. Proc. Natl Acad. Sci. USA 102, 8281–8286 (2005)

  31. 31.

    et al. Testing gene function early in the B cell lineage in mb1-cre mice. Proc. Natl Acad. Sci. USA 103, 13789–13794 (2006)

  32. 32.

    et al. Enforced bcl2 expression in B-lymphoid cells prolongs antibody responses and elicits autoimmune disease. Proc. Natl Acad. Sci. USA 88, 8661–8665 (1991)

  33. 33.

    , , & EBI2 mediates B cell segregation between the outer and centre follicle. Nature 460, 1122–1126 (2009)

  34. 34.

    et al. A loss-of-function RNA interference screen for molecular targets in cancer. Nature 441, 106–110 (2006)

  35. 35.

    et al. Balanced responsiveness to chemoattractants from adjacent zones determines B-cell position. Nature 416, 94–99 (2002)

  36. 36.

    et al. Lymphocyte egress from thymus and peripheral lymphoid organs is dependent on S1P receptor 1. Nature 427, 355–360 (2004)

  37. 37.

    , & Conversion of mature B cells into T cells by dedifferentiation to uncommitted progenitors. Nature 449, 473–477 (2007)

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We thank S. Coughlin for Gna13f/f and Arhgef1−/− mice and R. Proia for S1pr2−/− mice. We thank X. Geng and G. Doitsch for assistance with processing of human tonsil, A. Reboldi for discussion, and T. Arnon and O. Bannard for reading the manuscript. J.R.M. is supported by a Fellow Award from the Leukemia & Lymphoma Society and by National Institutes of Health (NIH) institutional training grants (T32 DK007636 and T32 CA1285835); R.S. is supported by the Dr Mildred Scheel Stiftung für Krebsforschung (Deutsche Krebshilfe). N.V. was supported by NIH grant GM097261 for the modelling work. J.G.C. is an Investigator of the Howard Hughes Medical Institute. The human lymphoma samples were studied under the auspices of the Lymphoma/Leukemia Molecular Profiling Project. The work was supported in part by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research, and NIH grant AI45073.

Author information

Author notes

    • Jesse A. Green

    Present address: Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.


  1. Department of Microbiology and Immunology, University of California, San Francisco, California, 94143, USA

    • Jagan R. Muppidi
    • , Jesse A. Green
    • , Sterling E. Braun
    • , Jinping An
    • , Ying Xu
    •  & Jason G. Cyster
  2. Department of Medicine, University of California, San Francisco, California 94143, USA

    • Jagan R. Muppidi
  3. Howard Hughes Medical Institute, University of California, San Francisco, California 94143, USA

    • Jagan R. Muppidi
    • , Jesse A. Green
    • , Sterling E. Braun
    • , Jinping An
    • , Ying Xu
    •  & Jason G. Cyster
  4. Lymphoid Malignancies Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Roland Schmitz
    • , Wenming Xiao
    •  & Louis M. Staudt
  5. Division of Immunology, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA

    • Adrien B. Larsen
    •  & Nagarajan Vaidehi
  6. Department of Pathology, University of Würzburg, 97080 Würzburg, Germany

    • Andreas Rosenwald
  7. Department of Clinical Pathology, Robert-Bosch-Krankenhaus, Auerbachstraße 110, 70376 Stuttgart, Germany

    • German Ott
  8. Dr. Margarete Fischer-Bosch Institute for Clinical Pharmacology, 70376 Stuttgart, Germany

    • German Ott
  9. British Columbia Cancer Agency, Vancouver, British Columbia V5Z 1L3, Canada

    • Randy D. Gascoyne
  10. Department of Pathology, University of Arizona, Tucson, Arizona 85724, USA

    • Lisa M. Rimsza
  11. Hospital Clinic, University of Barcelona, 08036 Barcelona, Spain

    • Elias Campo
  12. Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Elaine S. Jaffe
  13. Pathology Clinic, Rikshospitalet University Hospital, 0372 Oslo, Norway

    • Jan Delabie
  14. Institute for Cancer Research, Rikshospitalet University Hospital, University of Oslo, 0310 Oslo, Norway

    • Erlend B. Smeland
  15. Center for Cancer Biomedicine, Faculty Division of the Norwegian Radium Hospital, University of Oslo, 0310 Oslo, Norway

    • Erlend B. Smeland
  16. Oregon Health and Science University, Portland, Oregon 97239, USA

    • Rita M. Braziel
  17. Cleveland Clinic Pathology and Laboratory Medicine Institute, Cleveland, Ohio 44195, USA

    • Raymond R. Tubbs
    •  & J. R. Cook
  18. Department of Pathology, City of Hope National Medical Center, Duarte, California 91010, USA

    • Dennis D. Weisenburger
  19. Department of Pathology, University of Nebraska Medical Center, Omaha, Nebraska 68198, USA

    • Wing C. Chan
  20. Department of Microbiology, University of Nebraska Medical Center, Omaha, Nebraska 68198, USA

    • Wing C. Chan


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J.R.M. designed and performed experiments, interpreted the results and wrote the manuscript. R.S. performed sequencing of cell lines and primary samples, and analysed data. J.A.G. designed experimental procedures used in the manuscript. W.X. analysed sequence data. A.B.L and N.V. performed computer modelling of S1PR2. S.E.B. performed western blots of S1PR2. J.A. performed mouse genotyping and cared for the mouse colony. Y.X. performed quantitative PCR. A.R., G.O., R.D.G., L.M.R., E.C., E.S.J., J.D., E.B.S., R.M.B., R.R.T., J.R.C., D.D.W. and W.C.C. supplied lymphoma patient samples or lines, and reviewed pathological and clinical data. L.M.S. coordinated human sequence analysis, analysed data and supervised research. J.G.C. designed experiments, supervised research and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Louis M. Staudt or Jason G. Cyster.

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