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CXCR4 signaling directs Igk recombination and the molecular mechanisms of late B lymphopoiesis

Nature Immunology volume 20, pages 1393–1403 (2019)Cite this article

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Abstract

In B lymphopoiesis, activation of the pre-B cell antigen receptor (pre-BCR) is associated with both cell cycle exit and Igk recombination. Yet how the pre-BCR mediates these functions remains unclear. Here, we demonstrate that the pre-BCR initiates a feed-forward amplification loop mediated by the transcription factor interferon regulatory factor 4 and the chemokine receptor C-X-C motif chemokine receptor 4 (CXCR4). CXCR4 ligation by C-X-C motif chemokine ligand 12 activates the mitogen-activated protein kinase extracellular-signal-regulated kinase, which then directs the development of small pre- and immature B cells, including orchestrating cell cycle exit, pre-BCR repression, Igk recombination and BCR expression. In contrast, pre-BCR expression and escape from interleukin-7 have only modest effects on B cell developmental transcriptional and epigenetic programs. These data show a direct and central role for CXCR4 in orchestrating late B cell lymphopoiesis. Furthermore, in the context of previous findings, our data provide a three-receptor system sufficient to recapitulate the essential features of B lymphopoiesis in vitro.

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Fig. 1: Location of proliferating and differentiating pre-B cells in the BM.
Fig. 2: Small pre-B cells are in intimate contact with CXCL12+ stroma.
Fig. 3: Cxcr4 is required for the development of small pre-B cells.
Fig. 4: Pre-B cell differentiation is directly regulated by CXCR4 signaling.
Fig. 5: CXCR4 signaling determines small pre-B cell identity.
Fig. 6: CXCR4 signaling sets the small pre-B cell epigenetic landscape.
Fig. 7: CXCR4 signaling is necessary for Igk recombination.
Fig. 8: ERK signaling downstream of CXCR4 enables pre-B cell differentiation.

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Data availability

The sequences, ATAC-Seq data for WT small pre-B cells, mb1-Cre+Cxcr4fl/fl (Cxcr4 knockout) small pre-B cells, +IL-7, −IL-7 and −IL-7+CXCL12 cultured WT pre-B cells, and −IL-7 and −IL-7+CXCL12 cultured Cxcr4−/− small pre-B cells, as well as all of the corresponding RNA-Seq data, including for +IL-7+CXCL12 and −IL-7+CXCL12+ERK-i cultured pre-B cells, are deposited in the GenBank database (accession number GSE129311).

References

  1. Clark, M. R., Mandal, M., Ochiai, K. & Singh, H. Orchestrating B cell lymphopoiesis through interplay of IL-7 receptor and pre-B cell receptor signalling. Nat. Rev. Immunol. 14, 69–80 (2014).

    Article  CAS  Google Scholar 

  2. Herzog, S., Reth, M. & Jumaa, H. Regulation of B-cell proliferation and differentiation by pre-B-cell receptor signalling. Nat. Rev. Immunol. 9, 195–205 (2009).

    Article  CAS  Google Scholar 

  3. Clark, M. R., Cooper, A. B., Wang, L. & Aifantis, I. The pre-B cell receptor in B cell development: recent advances, persistent questions and conserved mechanisms. Curr. Top. Microbiol. Immunol. 290, 87–104 (2005).

    CAS  PubMed  Google Scholar 

  4. Cooper, A. B. et al. A unique function for cyclin D3 in early B cell development. Nat. Immunol. 7, 489–497 (2006).

    Article  CAS  Google Scholar 

  5. Lazorchak, A. S. et al. Sin1-mTORC2 suppresses rag and il7r gene expression through Akt2 in B cells. Mol. Cell 39, 433–443 (2010).

    Article  CAS  Google Scholar 

  6. Rathmell, J. C. et al. Akt-directed glucose matabolism can prevent Bax conformation change and promote growth factor-independent survival. Mol. Cell Biol. 23, 7315–7328 (2003).

    Article  CAS  Google Scholar 

  7. Gottlob, K. et al. Inhibition of early apoptotic events by Akt/PKB is dependent on the first committed step of glycolysis and mitochondrial hexokinase. Genes Dev. 15, 1406–1418 (2001).

    Article  CAS  Google Scholar 

  8. Mandal, M. et al. Ras orchestrates exit from the cell cycle and light-chain recombination during early B cell development. Nat. Immunol. 10, 1110–1117 (2009).

    Article  CAS  Google Scholar 

  9. Mandal, M. et al. Epigenetic repression of the Igk locus by STAT5-mediated recruitment of the histone methyltransferase Ezh2. Nat. Immunol. 12, 1212–1220 (2011).

    Article  CAS  Google Scholar 

  10. Mandal, M. et al. Histone reader BRWD1 targets and restricts recombination to the Igk locus. Nat. Immunol. 16, 1094–1103 (2015).

    Article  CAS  Google Scholar 

  11. Herzog, S. et al. SLP-65 regulates immunoglobulin light chain gene recombination through the PI(3)K-PKB-Foxo pathway. Nat. Immunol. 9, 623–631 (2008).

    Article  CAS  Google Scholar 

  12. Amin, R. H. & Schlissel, M. S. Foxo1 directly regulates the transcription of recombination-activating genes during B cell development. Nat. Immunol. 9, 613–622 (2008).

    Article  CAS  Google Scholar 

  13. Heizmann, B., Kastner, P. & Chan, S. Ikaros is absolutely required for pre-B cell differentiation by attenuating IL-7 signals. J. Exp. Med. 210, 2823–2832 (2013).

    Article  CAS  Google Scholar 

  14. Lin, Y. C. et al. A global network of transcription factors, involving E2A, EBF1 and Foxo1, that orchestrates B cell fate. Nat. Immunol. 11, 635–643 (2010).

    Article  CAS  Google Scholar 

  15. Sakamoto, S. et al. E2A and CBP/p300 act in synergy to promote chromatin accessibility of the immunoglobulin κ locus. J. Immunol. 188, 5547–5560 (2012).

    Article  CAS  Google Scholar 

  16. Van Loo, P. F., Dingjan, G. M., Maas, A. & Hendriks, R. W. Surrogate-light-chain silencing is not critical for the limitation of pre-B cell expansion but is for the termination of constitutive signaling. Immunity 27, 468–480 (2007).

    Article  CAS  Google Scholar 

  17. Fistonich, C. et al. Cell circuits between B cell progenitors and the IL-7+ mesenchymal progenitor cells control B cell development. J. Exp. Med. 215, 2596–2599 (2018).

    Article  Google Scholar 

  18. Tokoyoda, K., Egawa, T., Sugiyama, T., Choi, B. I. & Nagasawa, T. Cellular niches controlling B lymphocyte behavior within bone marrow during development. Immunity 20, 335–344 (2004).

    Article  Google Scholar 

  19. Johnson, K. et al. Regulation of immunoglobulin light-chain recombination by the transcription factor IRF-4 and the attenuation of interleukin-7 signaling. Immunity 28, 335–345 (2008).

    Article  CAS  Google Scholar 

  20. Tian, Y. et al. CXCL12 induces migration of oligodendrocyte precursor cells through the CXCR4-activated MEK/ERK and PI3K/AKT pathways. Mol. Med. Rep. 18, 4374–4380 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. Song, Z. Y., Wang, F., Cui, S. X., Gao, Z. H. & Qu, X. J. CXCR7/CXCR4 heterodimer-induced histone demethylation: a new mechanism of colorectal tumorigenesis. Oncogene 38, 1560–1575 (2019).

    Article  CAS  Google Scholar 

  22. Pozzobon, T., Goldoni, G., Viola, A. & Molon, B. CXCR4 signaling in health and disease. Immunol. Lett. 177, 6–15 (2016).

    Article  CAS  Google Scholar 

  23. Zheng, N. et al. CXCR7 is not obligatory for CXCL12-CXCR4-induced epithelial-mesenchymal transition in human ovarian cancer. Mol. Carcinog. 58, 144–155 (2019).

    Article  CAS  Google Scholar 

  24. Chatterjee, S., Behnam Azad, B. & Nimmagadda, S. The intricate role of CXCR4 in cancer. Adv. Cancer Res. 124, 31–82 (2014).

    Article  CAS  Google Scholar 

  25. Trampont, P. C. et al. CXCR4 acts as a costimulator during thymic β-selection. Nat. Immunol. 11, 162–170 (2010).

    Article  CAS  Google Scholar 

  26. Amin, R. H. et al. Biallelic, ubiquitous transcription from the distal germline Igκ locus promoter during B cell development. Proc. Natl Acad. Sci. USA 106, 522–527 (2009).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  28. Derudder, E. et al. Development of immunoglobulin λ-chain-positive B cells, but not editing of immunoglobulin κ-chain, depends on NF-κB signals. Nat. Immunol. 10, 647–454 (2009).

    Article  CAS  Google Scholar 

  29. Ochiai, K. et al. A self-reinforcing regulatory network triggered by limiting IL-7 activates pre-BCR signaling and differentiation. Nat. Immunol. 13, 300–307 (2012).

    Article  CAS  Google Scholar 

  30. Lagergren, A. et al. The Cxcl12, periostin, and Ccl9 genes are direct targets for early B-cell factor in OP-9 stroma cells. J. Biol. Chem. 282, 14454–14462 (2007).

    Article  CAS  Google Scholar 

  31. Mandal, M. et al. BRWD1 orchestrates epigenetic landscape of late B lymphopoiesis. Nat. Commun. 9, 3888–3902 (2018).

    Article  Google Scholar 

  32. Essafi, A. et al. Direct transcriptional regulation of Bim by FoxO3a mediates STI571-induced apoptosis in Bcr-Abl-expressing cells. Oncogene 24, 2317–2329 (2005).

    Article  CAS  Google Scholar 

  33. Nodland, S. E. et al. IL-7R expression and IL-7 signaling confer a distinct phenotype on developing human B-lineage cells. Blood 118, 2116–2127 (2011).

    Article  CAS  Google Scholar 

  34. 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).

    Article  CAS  Google Scholar 

  35. Bossen, C. et al. The chromatin remodeler Brg1 activates enhancer repertoires to establish B cell identity and modulate cell growth. Nat. Immunol. 16, 775–784 (2015).

    Article  CAS  Google Scholar 

  36. Montresor, A. et al. CXCR4- and BCR-triggered integrin activation in B-cell chronic lymphocytic leukemia cells depends on JAK2-activated Bruton’s tyrosine kinase. Oncotarget 9, 35123–35140 (2018).

    Article  Google Scholar 

  37. Thompson, E. C. et al. Ikaros DNA-binding proteins as integral components of B cell developmental-stage-specific regulatory circuits. Immunity 26, 335–344 (2007).

    Article  CAS  Google Scholar 

  38. Vikstrom, I. B. et al. MCL-1 is required throughout B-cell development and its loss sensitizes specific B-cell subsets to inhibition of BCL-2 or BCL-XL. Cell Death Dis. 7, e2345 (2016).

    Article  CAS  Google Scholar 

  39. Ma, S. et al. Ikaros and Aiolos inhibit pre-B cell proliferation by directly suppressing c-Myc expression. Mol. Cell Biol. 30, 4149–4158 (2010).

    Article  CAS  Google Scholar 

  40. Bolger, A. M., Lohse, M. & Usadel, B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120 (2014).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank M. Olson, R. Ladd and D. Leclerc for cell-sorting services, and the ImmGen Consortium for data assembly. We thank H. Singh (University of Pittsburgh) for providing the PAX5 and FOXO1 ChIP-Seq data, and M. Schlissel (University of Michigan) for providing the Cκ-YFP reporter mice. This work is supported by US National Institutes of Health grants AI120715, AI128785 and AI143778 (to M.R.C.), AI120715-02 and AI128785-01A1 (to M.M.), F32AI143120 (to D.E.K.), T32GM007281 (to K.C.M.), UL1TR002003 (to M.M.-C.) and T32HD007009 (to M.K.O.). Part of the bioinformatics analysis was performed by the UIC Research Informatics Core, supported in part by NCATS through grant UL1TR002003.

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Author notes

  1. These authors contributed equally: Michael K. Okoreeh, Domenick E. Kennedy.

Authors and Affiliations

  1. Department of Medicine, Section of Rheumatology, University of Chicago, Chicago, IL, USA

    Malay Mandal, Michael K. Okoreeh, Domenick E. Kennedy, Junting Ai, Kaitlin C. McLean, Margaret Veselits, Fotini Gounari & Marcus R. Clark

  2. Gwen Knapp Center for Lupus and Immunology Research, University of Chicago, Chicago, IL, USA

    Malay Mandal, Michael K. Okoreeh, Domenick E. Kennedy, Junting Ai, Kaitlin C. McLean, Margaret Veselits, Fotini Gounari & Marcus R. Clark

  3. Core for Research Informatics, University of Illinois at Chicago, Chicago, IL, USA

    Mark Maienschein-Cline

  4. Division of Nephrology, University of Washington, Seattle, WA, USA

    Natalya Kaverina

  5. Department of Pathology, NYU School of Medicine, New York, NY, USA

    Iannis Aifantis

  6. Laura and Isaac Perlmutter Cancer Center, NYU School of Medicine, New York, NY, USA

    Iannis Aifantis

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Contributions

M.M. and M.R.C. designed the experiments. M.M. carried out and analyzed most of the experiments. M.M. and M.M.-C. analyzed the high-throughput sequencing data. M.K.O and D.E.K. assisted with the ATAC-Seq, RNA-Seq and ChIP-Seq experiments, data analyses and flow cytometry. M.V. and K.C.M. performed some sorting and real-time PCR. J.I. and N.K. assisted with the confocal microscopy. I.A. helped to design some of the experiments. F.G. provided the RaDR-GFP mice. M.M. and M.R.C. oversaw the entire project and wrote the final manuscript.

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Correspondence to Malay Mandal or Marcus R. Clark.

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Peer review information: Laurie Dempsey was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

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Mandal, M., Okoreeh, M.K., Kennedy, D.E. et al. CXCR4 signaling directs Igk recombination and the molecular mechanisms of late B lymphopoiesis. Nat Immunol 20, 1393–1403 (2019). https://doi.org/10.1038/s41590-019-0468-0

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