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BACH2 mediates negative selection and p53-dependent tumor suppression at the pre-B cell receptor checkpoint

Abstract

The B cell–specific transcription factor BACH2 is required for affinity maturation of B cells. Here we show that Bach2-mediated activation of p53 is required for stringent elimination of pre-B cells that failed to productively rearrange immunoglobulin VH-DJH gene segments. After productive VH-DJH gene rearrangement, pre-B cell receptor signaling ends BACH2-mediated negative selection through B cell lymphoma 6 (BCL6)-mediated repression of p53. In patients with pre-B acute lymphoblastic leukemia, the BACH2-mediated checkpoint control is compromised by deletions, rare somatic mutations and loss of its upstream activator, PAX5. Low levels of BACH2 expression in these patients represent a strong independent predictor of poor clinical outcome. In this study, we demonstrate that Bach2+/+ pre-B cells resist leukemic transformation by Myc through Bach2-dependent upregulation of p53 and do not initiate fatal leukemia in transplant-recipient mice. Chromatin immunoprecipitation sequencing and gene expression analyses carried out by us revealed that BACH2 competes with BCL6 for promoter binding and reverses BCL6-mediated repression of p53 and other cell cycle checkpoint–control genes. These findings identify BACH2 as a crucial mediator of negative selection at the pre-B cell receptor checkpoint and a safeguard against leukemogenesis.

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Figure 1: Bach2 and Bcl6 maintain a balance between negative selection and survival of early B cells at the pre-B cell receptor checkpoint.
Figure 2: Bach2-dependent activation of Arf and p53 is reversed by Bcl6 as a result of expression of functional μHC.
Figure 3: Bach2 mediates V(D)J recombination and μHC checkpoint control during early B cell development.
Figure 4: BACH2 mediates PAX5-dependent tumor suppression in pre-B ALL through activation of TP53.
Figure 5: BACH2 is an independent predictor of poor clinical outcome in patients with ALL.
Figure 6: Bach2 prevents leukemic transformation by Myc.

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References

  1. 1

    Osmond, D.G. Proliferation kinetics and the lifespan of B cells in central and peripheral lymphoid organs. Curr. Opin. Immunol. 3, 179–185 (1991).

    CAS  Article  Google Scholar 

  2. 2

    Sakaguchi, N. & Melchers, F. Lambda 5, a new light-chain–related locus selectively expressed in pre-B lymphocytes. Nature 324, 579–582 (1986).

    CAS  Article  Google Scholar 

  3. 3

    Kitamura, D., Roes, J., Kühn, R. & Rajewsky, K.A. B cell–deficient mouse by targeted disruption of the membrane exon of the immunoglobulin mu chain gene. Nature 350, 423–426 (1991).

    CAS  Article  Google Scholar 

  4. 4

    Duy, C. et al. BCL6 is critical for the development of a diverse primary B cell repertoire. J. Exp. Med. 207, 1209–1221 (2010).

    CAS  Article  Google Scholar 

  5. 5

    Nahar, R. et al. Pre-B cell receptor-mediated activation of BCL6 induces pre-B cell quiescence through transcriptional repression of MYC. Blood 118, 4174–4178 (2011).

    CAS  Article  Google Scholar 

  6. 6

    van Zelm, M.C. et al. Ig gene rearrangement steps are initiated in early human precursor B cell subsets and correlate with specific transcription factor expression. J. Immunol. 175, 5912–5922 (2005).

    CAS  Article  Google Scholar 

  7. 7

    Fuxa, M. et al. Pax5 induces V-to-DJ rearrangements and locus contraction of the Ig heavy-chain gene. Genes Dev. 18, 411–422 (2004).

    CAS  Article  Google Scholar 

  8. 8

    Oyake, T. et al. Bach proteins belong to a novel family of BTB-basic leucine zipper transcription factors that interact with MafK and regulate transcription through the NF-E2 site. Mol. Cell Biol. 16, 6083–6095 (1996).

    CAS  Article  Google Scholar 

  9. 9

    Muto, A. et al. The transcriptional programme of antibody class switching involves the repressor Bach2. Nature 429, 566–571 (2004).

    CAS  Article  Google Scholar 

  10. 10

    Duy, C. et al. BCL6 enables Ph+ acute lymphoblastic leukaemia cells to survive BCR-ABL1 kinase inhibition. Nature 473, 384–388 (2011).

    CAS  Article  Google Scholar 

  11. 11

    Rouault, J.P. et al. Identification of BTG2, an antiproliferative p53-dependent component of the DNA damage cellular response pathway. Nat. Genet. 14, 482–486 (1996).

    CAS  Article  Google Scholar 

  12. 12

    Muljo, S.A. & Schlissel, M.S. A small molecule Abl kinase inhibitor induces differentiation of Abelson virus–transformed pre-B cell lines. Nat. Immunol. 4, 31–37 (2003).

    CAS  Article  Google Scholar 

  13. 13

    Malin, S. et al. Role of STAT5 in controlling cell survival and immunoglobulin gene recombination during pro-B cell development. Nat. Immunol. 11, 171–179 (2010).

    CAS  Article  Google Scholar 

  14. 14

    Ehlich, A., Martin, V., Müller, W. & Rajewsky, K. Analysis of the B-cell progenitor compartment at the level of single cells. Curr. Biol. 4, 573–583 (1994).

    CAS  Article  Google Scholar 

  15. 15

    Mullighan, C.G. et al. Genome-wide analysis of genetic alterations in acute lymphoblastic leukaemia. Nature 446, 758–764 (2007).

    CAS  Article  Google Scholar 

  16. 16

    Geng, H. et al. Integrative epigenomic analysis identifies biomarkers and therapeutic targets in adult B-acute lymphoblastic leukemia. Cancer Discov. 2, 1004–1023 (2012).

    CAS  Article  Google Scholar 

  17. 17

    Kawamata, N., Pennella, M.A., Woo, J.L., Berk, A.J. & Koeffler, H.P. Dominant-negative mechanism of leukemogenic PAX5 fusions. Oncogene 31, 966–977 (2012).

    CAS  Article  Google Scholar 

  18. 18

    Mullighan, C.G. et al. Deletion of IKZF1 and prognosis in acute lymphoblastic leukemia. N. Engl. J. Med. 360, 470–480 (2009).

    CAS  Article  Google Scholar 

  19. 19

    Merup, M. et al. 6q deletions in acute lymphoblastic leukemia and non-Hodgkin's lymphomas. Blood 91, 3397–3400 (1998).

    CAS  Article  Google Scholar 

  20. 20

    Hoelbl, A. et al. Stat5 is indispensable for the maintenance of bcr/abl-positive leukaemia. EMBO Mol. Med. 2, 98–110 (2010).

    CAS  Article  Google Scholar 

  21. 21

    Drayton, S. et al. Tumor suppressor p16INK4a determines sensitivity of human cells to transformation by cooperating cellular oncogenes. Cancer Cell 4, 301–310 (2003).

    CAS  Article  Google Scholar 

  22. 22

    Schmitt, C.A., McCurrach, M.E., de Stanchina, E., Wallace-Brodeur, R.R. & Lowe, S.W. INK4a/ARF mutations accelerate lymphomagenesis and promote chemoresistance by disabling p53. Genes Dev. 13, 2670–2677 (1999).

    CAS  Article  Google Scholar 

  23. 23

    Zindy, F. et al. Myc signalling via the ARF tumor suppressor regulates p53-dependent apoptosis and immortalization. Genes Dev. 12, 2424–2433 (1998).

    CAS  Article  Google Scholar 

  24. 24

    Trageser, D. et al. Pre-B cell receptor-mediated cell cycle arrest in Philadelphia chromosome-positive acute lymphoblastic leukemia requires IKAROS function. J. Exp. Med. 206, 1739–1753 (2009).

    CAS  Article  Google Scholar 

  25. 25

    Plaisier, S.B., Taschereau, R., Wong, J.A. & Graeber, T.G. Rank-rank hypergeometric overlap: identification of statistically significant overlap between gene-expression signatures. Nucleic Acids Res. 38, e169 (2010).

    Article  Google Scholar 

  26. 26

    Subramanian, A. et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. USA 102, 15545–15550 (2005).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We would like to thank H. Ye (Albert Einstein College of Medicine) for sharing Bcl6−/− mice and wild-type controls and L. Hennighausen (US National Institute of Diabetes and Digestive and Kidney Diseases) for Stat5flox/flox mice. Samples used in this research include those obtained from the Newcastle Haematology Biobank (http://www.ncl.ac.uk/nbb/collections/nhb/index.htm). We thank D.B. Kohn (University of California Los Angeles) for providing us with the retroviral envelope and packaging vectors used in this study. This work is supported by grants from the US National Institutes of Health National Cancer Institute through R01CA137060, R01CA139032, R01CA157644, R01CA172558, R01CA172558 and R01CA169458 (to M.M.), Translational Research Program grants from the Leukemia and Lymphoma Society (grants 6132-09 and 6097-10), a Leukemia and Lymphoma Society Specialized Center of Research (grant 7005-11, B.J. Druker (principal investigator)), the William Lawrence and Blanche Hughes Foundation and a Stand Up To Cancer–American Association for Cancer Research Innovative Research Grant (IRG00909 to M.M.), the California Institute for Regenerative Medicine (CIRM; TR2-01816 to M.M.) and Leukaemia and Lymphoma Research (V.R. and A.G.H.). A.M. and M.M. are Scholars of the Leukemia and Lymphoma Society. V.R. is a Leukaemia and Lymphoma Research Bennett Fellow.

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S.S. and M.M. designed experiments and interpreted data. M.M. and S.S. also conceived the study and wrote the paper. S.S., C. Huang, H.G., Z.C., C.N., B.T., C. Hurtz, M.F.S., D.N., G.B.T. and V.R. performed experiments and interpreted data. H.G., R.H., H.K., V.R., H.P.K., W.L.C., C.L.W., A.G.H. and A.M. provided and characterized patient samples and clinical outcome data. T.G.G., H.P.K., K.I. and A.M. provided important reagents and mouse samples.

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Correspondence to Markus Müschen.

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The authors declare no competing financial interests.

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Swaminathan, S., Huang, C., Geng, H. et al. BACH2 mediates negative selection and p53-dependent tumor suppression at the pre-B cell receptor checkpoint. Nat Med 19, 1014–1022 (2013). https://doi.org/10.1038/nm.3247

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