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EBF1 and PAX5 control pro-B cell expansion via opposing regulation of the Myc gene

Somasundaram, Rajesh and Jensen, Christina T. and Tingvall-Gustafsson, Johanna and Åhsberg, Josefine and Okuyama, Kazuki and Prasad, Mahadesh and Hagman, James R. and Wang, Xun and Soneji, Shamit and Strid, Tobias and Ungerbäck, Jonas and Sigvardsson, Mikael (2021) EBF1 and PAX5 control pro-B cell expansion via opposing regulation of the Myc gene. Blood, 137 (22). pp. 3037-3049. ISSN 0006-4971. doi:10.1182/blood.2020009564.

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Genes encoding B lineage–restricted transcription factors are frequently mutated in B-lymphoid leukemias, suggesting a close link between normal and malignant B-cell development. One of these transcription factors is early B-cell factor 1 (EBF1), a protein of critical importance for lineage specification and survival of B-lymphoid progenitors. Here, we report that impaired EBF1 function in mouse B-cell progenitors results in reduced expression of Myc. Ectopic expression of MYC partially rescued B-cell expansion in the absence of EBF1 both in vivo and in vitro. Using chromosome conformation analysis in combination with ATAC-sequencing, chromatin immunoprecipitation–sequencing, and reporter gene assays, six EBF1-responsive enhancer elements were identified within the Myc locus. CRISPR-Cas9–mediated targeting of EBF1-binding sites identified one element of key importance for Myc expression and pro-B cell expansion. These data provide evidence that Myc is a direct target of EBF1. Furthermore, chromatin immunoprecipitation–sequencing analysis revealed that several regulatory elements in the Myc locus are targets of PAX5. However, ectopic expression of PAX5 in EBF1-deficient cells inhibits the cell cycle and reduces Myc expression, suggesting that EBF1 and PAX5 act in an opposing manner to regulate Myc levels. This hypothesis is further substantiated by the finding that Pax5 inactivation reduces requirements for EBF1 in pro–B-cell expansion. The binding of EBF1 and PAX5 to regulatory elements in the human MYC gene in a B-cell acute lymphoblastic leukemia cell line indicates that the EBF1:PAX5:MYC regulatory loop is conserved and may control both normal and malignant B-cell development.

Item Type:Article
Related URLs:
URLURL TypeDescription
Tingvall-Gustafsson, Johanna0000-0002-7359-6584
Hagman, James R.0000-0002-5436-8455
Wang, Xun0000-0002-1653-5750
Strid, Tobias0000-0002-2166-5170
Ungerbäck, Jonas0000-0002-2190-3896
Sigvardsson, Mikael0000-0001-8527-7276
Alternate Title:Opposing regulation of Myc by EBF1 and PAX5
Additional Information:© 2021 American Society of Hematology. The authors are grateful for the technical assistance provided by Liselotte Lenner, Linda Bergström, and Maria Malmberg. This work was supported by grants from the Swedish Cancer Society (2017-258), the Swedish Childhood Cancer Foundation (2019-0020), the Swedish Research Council (2018-02448), including a Strategic research grant to Stem Therapy, Knut and Alice Wallenberg’s Foundation (2014-0089), and a donation from Henry Hallberg (all, M.S.) and Lions forskningsfond mot folksjukdomar (T.S.). J.R.H. is funded by the National Institutes of Health, National Institute of Allergy and Infectious Diseases (R21AI115696), and by the Wendy Siegel Fund for Leukemia and Cancer Research. Sequencing data are deposited in the GEO database as accession number GSE136238 (supplemental Figure 1; supplemental Data sheets) and accession number GSE159957 (Figures 3 and 5). Authorship Contribution: K.O., T.S., R.S., C.T.J., J.T.-G., M.P., J.Å., M.S., and J.U. designed, conducted, and analyzed experiments; S.S., J.U., C.T.J., and T.S. contributed to the bioinformatics analysis; J.R.H. and X.W. contributed essential reagents and analyzed data; M.S. designed experiments, analyzed data, and wrote the manuscript draft; and all authors contributed to the final version of the manuscript. R.S. and C.T.J. contributed equally to this study. The authors declare no competing financial interests. The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked “advertisement” in accordance with 18 USC section 1734.
Funding AgencyGrant Number
Swedish Cancer Society2017-258
Swedish Childhood Cancer Foundation2019-0020
Swedish Research Council2018-02448
Knut and Alice Wallenberg Foundation2014-0089
Lions forskningsfond mot folksjukdomarUNSPECIFIED
Wendy Siegel Fund for Leukemia and Cancer ResearchUNSPECIFIED
Subject Keywords:b-lymphocytes, cell growth, c-myc genes, mice, pax5 gene, chromatin immunoprecipitation sequencing, genes, myc, burkitt's lymphoma, atac trial, transcription factor
Issue or Number:22
Record Number:CaltechAUTHORS:20210224-125024033
Persistent URL:
Official Citation:Rajesh Somasundaram, Christina T. Jensen, Johanna Tingvall-Gustafsson, Josefine Åhsberg, Kazuki Okuyama, Mahadesh Prasad, James R. Hagman, Xun Wang, Shamit Soneji, Tobias Strid, Jonas Ungerbäck, Mikael Sigvardsson; EBF1 and PAX5 control pro-B cell expansion via opposing regulation of the Myc gene. Blood 2021; 137 (22): 3037–3049. doi:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:108175
Deposited By: Tony Diaz
Deposited On:24 Feb 2021 21:07
Last Modified:03 Jun 2021 22:08

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