Chronic active B-cell-receptor signalling in diffuse large B-cell lymphoma

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Abstract

A role for B-cell-receptor (BCR) signalling in lymphomagenesis has been inferred by studying immunoglobulin genes in human lymphomas1,2 and by engineering mouse models3, but genetic and functional evidence for its oncogenic role in human lymphomas is needed. Here we describe a form of ‘chronic active’ BCR signalling that is required for cell survival in the activated B-cell-like (ABC) subtype of diffuse large B-cell lymphoma (DLBCL). The signalling adaptor CARD11 is required for constitutive NF-κB pathway activity and survival in ABC DLBCL4. Roughly 10% of ABC DLBCLs have mutant CARD11 isoforms that activate NF-κB5, but the mechanism that engages wild-type CARD11 in other ABC DLBCLs was unknown. An RNA interference genetic screen revealed that a BCR signalling component, Bruton’s tyrosine kinase, is essential for the survival of ABC DLBCLs with wild-type CARD11. In addition, knockdown of proximal BCR subunits (IgM, Ig-κ, CD79A and CD79B) killed ABC DLBCLs with wild-type CARD11 but not other lymphomas. The BCRs in these ABC DLBCLs formed prominent clusters in the plasma membrane with low diffusion, similarly to BCRs in antigen-stimulated normal B cells. Somatic mutations affecting the immunoreceptor tyrosine-based activation motif (ITAM) signalling modules6 of CD79B and CD79A were detected frequently in ABC DLBCL biopsy samples but rarely in other DLBCLs and never in Burkitt’s lymphoma or mucosa-associated lymphoid tissue lymphoma. In 18% of ABC DLBCLs, one functionally critical residue of CD79B, the first ITAM tyrosine, was mutated. These mutations increased surface BCR expression and attenuated Lyn kinase, a feedback inhibitor of BCR signalling. These findings establish chronic active BCR signalling as a new pathogenetic mechanism in ABC DLBCL, suggesting several therapeutic strategies.

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Figure 1: BTK is a critical kinase for survival of ABC DLBCL cells.
Figure 2: Chronic active BCR signalling in ABC DLBCL lines.
Figure 3: CD79A and CD79B ITAM mutations in ABC DLBCL.
Figure 4: Therapeutic strategies to target chronic active BCR signalling.

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Primary accessions

Gene Expression Omnibus

Data deposits

Gene expression profiling data have been deposited in the Gene Expression Omnibus (GEO) under accession number GSE18817.

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Acknowledgements

We thank L. Honigberg for PCI-32765, L. Dang for IKK-β inhibitors, S. Ansher for dasatinib, and S. Tohda for the TMD8 cell line. This research was supported by the Intramural Research Program of the National Institutes of Health, the National Cancer Institute, the Center for Cancer Research, the National Institute of Allergy and Infectious Disease, and the National Human Genome Research Institute. P.B.R. was a Howard Hughes Medical Institute-National Institutes of Health Research Scholar.

Author Contributions R.E.D., V.N.N., G.L., P.T., R.M.Y., P.B.R., H.K., L.L. and A.L.S. designed and performed experiments. H.Z., Y.Y. and W.X. performed experiments. G.W., W.X. and J.P. analysed data. J.J. and C.J.T. synthesized reagents. A.R., G.O., H.K.M.-H., R.D.G., J.M.C., N.A.J., L.M.R., E.C., E.S.J., W.H.W., J.D., E.B.S., R.I.F., R.M.B., R.R.T., J.R.C., D.D.W. and W.C.C. supplied samples from patients and reviewed pathological and clinical data. S.K.P. supervised research. L.M.S. designed and supervised research and wrote the manuscript.

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Correspondence to Louis M. Staudt.

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